Table_1_Effects of secukinumab and ixekizumab on major adverse cardiovascular events in patients with psoriasis: a meta-analysis of randomized controlled trials.docx

IntroductionThe aims of this study is to analyze the risk of major adverse cardiovascular events (MACEs) in patients with psoriasis treated with secukinumab and ixekizumab.MethodologyWe systematically identified randomized controlled trials (RCTs) that focused on the treatment of psoriasis with secukinumab and ixekizumab by conducting computerized searches of PubMed, Embase, and the Cochrane Library databases, spanning from their inception to October 31st, 2022. The search terms used included psoriasis, secukinumab, ixekizumab, and randomized controlled trial. Two independent evaluators conducted literature screening, data extraction, and assessed the quality of included studies based on predetermined inclusion and exclusion criteria. The gather data was subjected to meta-analysis using the statistical software RevMan 5.4.ResultsA total of 20 articles, encompassing 23 randomized controlled trials involving 10,746 psoriasis patients were included in the analysis. During the double-blind treatment period, the meta-analysis results indicated the following: There was no significant difference in the incidence of MACEs between the secukinumab and placebo groups [RR = 0.61, 95% CI (0.26, 1.44), p = 0.26]. Similarly, there was no significant difference in the incidence of MACEs with ixekizumab compared to the placebo group [RR = 0.47, 95% CI (0.15, 1.47), p = 0.20]. Furthermore, no significant difference in the incidence of MACEs was observed between secukinumab 300 mg and secukinumab 150 mg treatment groups [RR = 1.00, 95% CI (0.23, 4.35), p = 1.00]. Likewise, there was no significant difference in the incidence of MACEs between the ixekizumab Q4W (every 4 weeks) and ixekizumab Q2W (every 2 weeks) administration groups [RR = 4.01, 95% CI (0.45, 35.89), p = 0.21].ConclusionThe findings of this study suggest that neither secukinumab nor ixekizumab is significantly associated with the risk of MACEs in patients with psoriasis during double-blind treatment.Systematic review registration: Unique Identifier: CRD42022373756 https://www.crd.york.ac.uk/.</p

Responses of mutants with glycerol metabolism defects to exogenous glycerol.

<p>(A) Detection of <i>GLI1</i>, <i>GPDHc1</i> and <i>FAD-GPDH</i> transcripts by RT-PCR in wild-type, <i>gli1</i>, <i>gpdhc1</i> and <i>fad-gpdh</i> seedlings. Total RNA was prepared from 3-day-old seedlings. <i>Actin07</i> was used as an internal control. Wild-type and <i>gli1, gpdhc1</i> and <i>fad-gpdh</i> mutant seedlings were grown for 6 days on vertically oriented agar plates. (B) Seedlings growing on media containing various concentrations of glycerol (0, 250 µM and 1 mM) are shown. Bar = 1 cm. (C) The primary root (PR) lengths of WT and <i>gli1</i>, <i>gpdhc1</i> and <i>fad-gpdh</i> mutant seedlings were recorded. (D) Data on the number of lateral root primordia (LRP) per plant were statistically analyzed. The data are presented as the mean of 30–40 seedlings ± SE. Asterisks indicate significant differences between the treatment (250 µM and 1 mM glycerol) and control (0) (*, p<0.05; **, p<0.01) by Student’s t-test.</p

Glycerol Affects Root Development through Regulation of Multiple Pathways in Arabidopsis

<div><p>Glycerol metabolism has been well studied biochemically. However, the means by which glycerol functions in plant development is not well understood. This study aimed to investigate the mechanism underlying the effects of glycerol on root development in <i>Arabidopsis thaliana</i>. Exogenous glycerol inhibited primary root growth and altered lateral root development in wild-type plants. These phenotypes appeared concurrently with increased endogenous glycerol-3-phosphate (G3P) and H₂O₂ contents in seedlings, and decreased phosphate levels in roots. Upon glycerol treatment, G3P level and root development did not change in glycerol kinase mutant <i>gli1</i>, but G3P level increased in <i>gpdhc1</i> and <i>fad-gpdh</i> mutants, which resulted in more severely impaired root development. Overexpression of the <i>FAD-GPDH</i> gene attenuated the alterations in G3P, phosphate and H₂O₂ levels, leading to increased tolerance to exogenous glycerol, which suggested that FAD-GPDH plays an important role in modulating this response. Free indole-3-acetic acid (IAA) content increased by 46%, and <i>DR5pro::GUS</i> staining increased in the stele cells of the root meristem under glycerol treatment, suggesting that glycerol likely alters normal auxin distribution. Decreases in <i>PIN1</i> and <i>PIN7</i> expression, <i>β</i>-glucuronidase (GUS) staining in plants expressing <i>PIN7pro::GUS</i> and green fluorescent protein (GFP) fluorescence in plants expressing <i>PIN7pro::PIN7-GFP</i> were observed, indicating that polar auxin transport in the root was downregulated under glycerol treatment. Analyses with auxin-related mutants showed that TIR1 and ARF7 were involved in regulating root growth under glycerol treatment. Glycerol-treated plants showed significant reductions in root meristem size and cell number as revealed by <i>CYCB1;1pro::GUS</i> staining. Furthermore, the expression of <i>CDKA</i> and <i>CYCB1</i> decreased significantly in treated plants compared with control plants, implying possible alterations in cell cycle progression. Our data demonstrated that glycerol treatment altered endogenous levels of G3P, phosphate and ROS, affected auxin distribution and cell division in the root meristem, and eventually resulted in modifications of root development.</p></div

G3P levels in seedlings treated with glycerol (nmol g⁻¹ FW).

<p>(A) Wild-type seedlings were grown on agar plates containing 0.5×Murashige and Skoog (MS) medium plus 1% (w/v) sucrose in the absence or presence of 1 mM glycerol from 1–5 days post-germination (dpg). The G3P levels of the seedlings were examined. The data are presented as the mean ± SE (n = 3–4). The asterisks indicate significant differences between the means as determined by Student’s t-test (control versus 1 mM glycerol: *, p<0.05; **, p<0.01). (B) Wild-type, <i>gli1, gpdhc1</i>, <i>fad-gpdh</i>, OE #16 and OE #22 seedlings were grown on agar plates containing 0.5×MS medium plus 1% (w/v) sucrose in the absence or presence of 1 mM glycerol for 4 days. The G3P levels were assayed. The data are presented as the mean ± SE (n = 3–6). The symbols indicate significant differences between the means (*: control versus 1 mM glycerol; #: WT versus mutants and OE lines. *, #: p<0.05; **, ##: p<0.01).</p

Exogenous glycerol inhibits Arabidopsis primary root growth and has variable effects on lateral root primordia.

<p>Wild-type seedlings were grown on plates containing 0.5×Murashige and Skoog (MS) medium with 1 mM glycerol for the indicated number of days. (A) Root development of seedlings grown on control (left) and 1 mM glycerol media (right) at 2–8 days post germination (dpg). Bar = 0.5 cm. (B) Quantification of the primary root (PR) lengths of the seedlings grown under the conditions described in (A). (C) Quantification of the lateral root primordia (LRPs) of the seedlings grown under the conditions described in (A). The developmental stage of each LRP was classified according to Zhang <i>et al.</i> (1999): Stage A, up to three cell layers; Stage B, unemerged LR, but more than three cell layers; Stage C, emerged LR <0.5 mm in length; Stage D, LR longer than 0.5 mm. The data are presented as the mean ± SE (n = 10). Asterisks indicate a significant difference at p<0.05 (*) or p<0.01 (**) by Student’s t-test.</p

Pi and H₂O₂ levels in seedlings treated with glycerol.

<p>Wild-type seedlings were grown on agar plates containing 0.5×MS medium plus 1% (w/v) sucrose in the absence or presence of 1 mM glycerol from 2–6 days post-germination (dpg). (A–B) The cellular Pi level (µmol g⁻¹ FW) was analyzed in wild-type roots (A) and shoots (B) for the indicated number of days. The data are presented as the mean ± SE (n = 4–6). (C–D) The cellular Pi level in the roots (C) and shoots (D) of the <i>gli1</i> mutant, OE #16, OE #22 and wild-type seedlings grown on 0 or 1 mM glycerol medium was assayed at 4 dpg. Values are expressed as the mean ± SE (n = 4). (E) The effect of phosphate availability on the primary root (PR) length in wild-type and <i>FAD-GPDH^OE</i> lines. Arabidopsis wild-type (Col-0), OE #16 and OE #22 seedlings were grown on the surface of agar plates containing various concentrations of phosphate for 6 days, and the PR length was measured. (F) Quantification of H₂O₂ in <i>gli1, gpdhc1</i> and <i>fad-gpdh</i> mutants, OE #16, OE #22 and wild-type seedlings grown on 0 or 1 mM glycerol medium was performed at 5 dpg. The values are expressed as the mean ± SE (n = 4). (G) The effect of exogenous H₂O₂ on PR length. PR lengths of 5-day-old <i>gli1</i> mutants, OE #16, OE #22 and wild-type seedlings in the presence of 0, 250 µM and 1 mM H₂O₂ were recorded. Values are expressed as the means ± SE (n = 18–20). Different symbols indicate that the means differ significantly by Student’s t-test (*: control versus 1 mM glycerol or other treatment; #: WT versus mutants or OE lines. *, #: p<0.05; **, ##: p<0.01).</p

A model illustrating glycerol-triggered modulation of root development.

<p>The diagram shows the different root patterns in the absence (left) or the presence (right) of glycerol. The middle section shows a condensed schematic of plant glycerol metabolism and three important genes in this study (red). Glycerol is phosphorylated to G3P by GLI1 and can also be generated by GPDHc1 via the reduction of DHAP. G3P is oxidized to DHAP by FAD-GPDH or dephosphorylate to glycerol by GPP. Exogenous glycerol treatment can cause modifications of multiple pathways, including increased G3P and reactive oxygen species (ROS) levels, reduced the phosphate level and expression of <i>PIN1</i> and <i>PIN7</i>. It also affected polar auxin transport and the root meristem activity, thus resulting in modified root growth and development. Abbreviations: GLI1, glycerol kinase; GPDHc1, cytosolic glycerol-3-phosphate dehydrogenase; FAD-GPDH, flavin adenine dinucleotide-dependent glycerol-3-phosphate dehydrogenase; GPP, glycerol-3-phosphatase; G3P, glycerol-3-phosphate; DHAP, dihydroxyacetone phosphate; ATP, adenosine triphosphate; FAD, flavin adenine dinucleotide; NADH, the reduced form of nicotinamide adenine dinucleotide. For the sake of clarity, some co-substrates and/or co-products have been omitted in some reactions.</p

The effect of auxin transport inhibitor NPA on root development in Arabidopsis seedlings.

<p>(A) Growth performance of wild-type seedlings under glycerol and NPA treatments. Wild-type seedlings were grown on 0.5×MS medium for 5 days and subsequently transferred to media containing 1 mM glycerol with or without 2 µM NPA for 3 days. Black dots indicated the starting growth positions of the PR tip after shift. Bar = 1 cm. (B) Primary root (PR) extension lengths and (C) Lateral root primordia (LRP) at Stages A and D in wild-type plants grown under the conditions as described in (A) were recorded. The data are presented as the mean ± SE (n = 36). (D) Comparison of wild-type and OE #16 plants under glycerol and NPA treatments. Wild-type and OE #16 seeds were sown directly onto media containing 0 mM glycerol, 1 mM glycerol and 2 µM NPA or 1 mM glycerol and 2 µM NPA for 7 days; the root lengths of the plants exposed to each treatment were then measured (n≥27). Different letters indicate significant differences (p<0.05).</p

Root meristem cell and cell cycle gene expression in glycerol-treated seedlings of wild-type and mutants.

<p>The seedlings were grown on 0.5×MS medium plus 1% sucrose with or without 1 mM glycerol. (A) Nomarski images showed the meristems of wild-type seedlings gown on 0.5×MS medium in the absence (left) or presence (right) of 1 mM glycerol at 8 dpg. Arrows mark the boundaries of the meristem region. Bars = 100 µm. The meristem size (B) and meristem cell number (C) of wild-type plants grown on media with or without glycerol at different developmental stages were investigated. Meristem size (D) and meristem cell number (E) of wild-type, <i>gpdhc1</i>, <i>gli1</i> and <i>fad-gpdh</i> seedlings grown for 7 days on 0, 250 µM and 1 mM glycerol media were recorded. The data are presented as the mean of 30–40 seedlings ± SE. (F) Starch granules in wild-type, <i>gli1</i>, <i>gpdhc1</i> and <i>fad-gpdh</i> seedlings were visualized by Lugol staining in the presence or absence of 1 mM glycerol. Four-day-old seedlings were first fixed in FAA at 4°C overnight and subsequently washed once in 50% ethanol. The samples were then placed in Lugol solution (0.37% iodine and 0.71% potassium iodide) for 1 min and transferred to a chloral hydrate solution for 2 min. The micrographs are representative of at least 10 seedlings for each genotype. Bars = 20 µm. (G) The expression levels of <i>CycB1</i> and <i>CDKA</i> in the root tip at 8 dpg were analyzed under glycerol treatment. (H) Seedlings expressing <i>CycB1;1-GUS</i> were grown on media in the presence or absence of 1 mM glycerol for 6 days and subjected to GUS staining. Bar = 10 µm. Asterisks indicate that the means differ significantly from the control by Student’s t-test (*, p<0.05; **, p<0.01).</p

The overexpression of <i>AtFAD-GPDH</i> in Arabidopsis ameliorates the effect of glycerol on root development.

<p>(A) Transgene expression levels in different transgenic lines. Reverse transcription PCR (RT-PCR) was performed on cDNA made from total RNA extracted from 7-day-old wild-type and <i>35Spro::AtFAD-GPDH</i> seedlings. <i>Actin07</i> is shown as an internal control. (B) Seven-day-old wild-type seedlings and four <i>35Spro::AtFAD-GPDH</i> lines were grown on agar plates containing 0.5×MS medium plus 1% (w/v) sucrose in the presence or absence of 1 mM glycerol. Bar = 0.5 cm. (C and D) Primary root (PR) length (C) and lateral root (LR) number (D) of 7-day-old wild-type and transgenic seedlings described in (B) were recorded. The data are presented as the mean ± SE (n≥20). (E) The relative expression of <i>FAD-GPDH</i> in the 8-day-old wild-type seedling roots under 1 mM glycerol treatment compared with untreated control. (F and G) The expression pattern of <i>FAD-GPDHpro:GUS</i> in germinating seeds (F); Bar = 100 µm, and 3-day-old seedlings (G) under normal growth conditions; Bar = 500 µm. At least five transgenic plants were observed at each stage, and representative images are presented. The arrows in (F) and (G) show GUS staining in the root cap.</p