Replication of <i>C. trachomatis</i> organisms in PRAK-deficient mouse embryo fibroblast cells.

<p>Mouse embryo fibroblast cells (MEF) without (panels a–d) or with (e–h) PRAK deficiency (PRAK−/−) were infected with (b–d & f–h) or without (a & e) <i>C. trachomatis</i> (MOI = 0.5) for various periods of time as indicated on top of the figure. The cultures were processed for immunofluorescence assay with a rabbit antibody for visualizing chlamydial organisms (green), Alexa-Fluor 568 Phalloidin for host cell F-actin (red) and Hoechst dye for DNA (blue). Note that the inclusion sizes were similar in MEF with or without PRAK deficiency.</p

Rottlerin inhibits chlamydial growth in the absence of PRAK.

<p>MEF without (panels a–c) or with (d–f) PRAK deficiency (PRAK−/−) were infected with <i>C. trachomatis</i> (MOI = 0.5) and at oh (b & e) or 16 h (c & f) post infection, parallel cultures were treated with (b, c, e & f) or without (a & d) rottlerin at 1 µM. The cultures were processed 44 h post infection for immunofluorescence assay as described in Fig. 3 legend. Note that rottlerin inhibited chlamydial growth in both wild type and PRAK-deficient MEF cells.</p

<i>Chlamydia trachomatis</i> acquisition of host sphingomyelin is independent of PRAK.

<p>(A) HeLa cells with (panels e–h) or without (a–d) <i>C. trachomatis</i> infection (MOI = 0.5) were treated without (a & e) or with EGCG (1 µM, b & f; 10 µM, c & g) or rottlerin (1 µM, d & h) 16 h post infection. Eight hours later, the cultures were subjected to BODIPY-FL-C5-ceremide labeling and visualized under a fluorescence microscope. Note that EGCG failed to block the accumulation of BODIPY-FL-sphingomyelin in the chlamydial inclusions (panel f & g) while rottlerin did (h). (B) MEF without (panels a & c) or with (b & d) PRAK deficiency (PRAK−/−) were infected with <i>C. trachomatis</i> (MOI = 0.5) and 24 h post infection, the cultures were labeled with BODIPY-FL-C5-ceremide and observed as described above. Note that <i>C. trachomatis</i> organisms can take up BODIPY-FL-sphingomyelin from MEF cells with or without PRAK. The thick arrows point to chlamydial inclusions with while thin arrows point to the inclusions without the fluorescent sphingomyelin.</p

EGCG fails to inhibit chlamydial growth.

<p>HeLa cells infected with <i>C. trachomatis</i> (MOI = 0.5) were treated with EGCG (1 µM, panel b & 10 µM, c) or rottlerin (1 µM, d) 16 h post infection. The cultures were processed 44 h post infection for immuno-labeling with a rabbit antibody for labeling the <i>C. trachomatis</i> organisms (green) and a mouse monoclonal antibody (clone BB2) for inclusion membrane protein IncA (red). Note that EGCG at 1 µM failed to inhibit inclusion expansion and even at 10 µM only slightly reduced inclusion size while rottlerin at 1 µM completely blocked inclusion expansion.</p

Ginger for treating nausea and vomiting: an overview of systematic reviews and meta-analyses

Ginger may be a potential remedy for nausea and vomiting. This review aimed to assess the reporting and methodological quality, and integrate the evidence in this field. A total of fifteen meta-analyses were analysed and met the Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2009 guidelines, providing a relatively complete statement. However, methodological quality, assessed using the Assessment of Multiple Systematic Reviews-2 checklist, was deemed critically low to low. Our review’s findings support ginger’s effectiveness in managing chemotherapy-induced nausea and vomiting in cancer patients. It also reduces postoperative nausea and vomiting severity, decreasing the need for rescue antiemetics. Furthermore, ginger shows promise in alleviating pregnancy-related nausea and vomiting symptoms. The pooled evidence suggests ginger as a safe botanical option for managing nausea and vomiting, but it is important to improve the scientific quality of published meta-analyses in the future.</p

Basic amino acids required for targeting GFP into nucleus.

<p>(A) The amino acid sequence covering residue 21 to 63 defined as CT311 fragment 4 (F4) or nuclear localization signal sequence (NLS) was shown with two clusters of basic residues highlighted in red and marked as Clusters 1 & 2 (C1 & C2) respectively. Cluster 2 consists of 2 separate basic residue sub-clusters, designated as C2a & C2b respectively. The sequences of constructs with C2b deletion (C2b del-GFP) and C1 or C2a substitution mutations (C1 sub-GFP or C2a sub-GFP) were also listed. (B) HeLa cells transfected with pLEGFP plasmid alone or expressing the various constructs listed in (A) were processed and observed under a fluorescence microscope as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064529#pone-0064529-g004" target="_blank">Fig. 4B</a> legend. Deletion or substitution of C2b, C1 or C2a effectively blocked the nuclear targeting of GFP by the CT311 NLS. (C) The GFP signal in the nuclei and entire cells was semi-quantitatively measured and the % of nuclear GFP signal from each sample was compared between cell samples as described in the legend of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064529#pone-0064529-g004" target="_blank">Fig4</a>. ** indicates statistically significant differences (P<0.01) and “ns” stands for no significant difference. The data were from 3 independent experiments.</p

Nuclear localization of CT311 in <i>C.</i><i>trachomatis</i>-infected cells.

<p>(A) HeLa cells infected with <i>C. trachomatis</i> organisms were processed at different time points after infection (as indicated on top of the image) for immunofluorescence labeling of chlamydial organisms (green), CT311 (panels a to e) or CPAF (panels f to j, both red) or DNA (blue). Red arrowheads indicate CT311 localized in the host cell nuclei in samples harvested at 36h (d) and 44h (e) post infection. The images were acquired using a conventional fluorescence microscope. (B) The 36h sample was further observed under a confocal microscope. Red arrowhead indicates nuclear localization of CT311.</p

Localization of CT311 expressed via a transgene in mammalian cell nuclei.

<p>(A) HeLa cells transfected with pLenti6.3/V5-CT311 (panels a–c) or pLenti6.3/V5-CPAF (panels d to f) were processed 24h after transfection for immunofluorescence labeling of CT311 or CPAF using an anti-V5 tag antibody (red) and DNA using Hoechst dye (blue). CT311 but not CPAF localized in host cell nuclei. (B) The nuclear localization of CT311 was confirmed by co-labeling CT311 (green) and Nup153 (red), a nuclear protein.</p

Table_3_Natural products and dietary interventions on liver enzymes: an umbrella review and evidence map.DOCX

BackgroundThe association between natural products and dietary interventions on liver enzymes is unclear; therefore, this study aimed to examine their effects on liver enzymes in adults.MethodsPubMed, Embase, and Cochrane Library of Systematic Reviews databases were searched from inception until March 2023. The Assessment of Multiple Systematic Reviews-2 (AMSTAR-2) and Grading of Recommendations Assessment, Development, and Evaluation (GRADE) systems were used to assess the methodological and evidence quality, and the therapeutic effects were summarized in a narrative form.ResultsA total of 40 meta-analyses on natural products (n = 25), dietary supplements (n = 10), and dietary patterns (n = 5) were evaluated, and results were presented in a narrative form. The overall methodological quality of the included studies was relatively poor. The results indicated that positive effects were observed for nigella sativa, garlic, artichoke, curcumin, silymarin, vitamin E, vitamin D, L-carnitine, propolis, and polyunsaturated fatty acids on certain liver enzymes. The dietary patterns, including high-protein, Mediterranean, and calorie-restriction diets and evening snacks, may reduce liver enzymes; however, other supplements and herbs did not reduce liver enzyme levels or have minimal effects. The evidence quality was generally weak given the risk of bias, heterogeneity, and imprecision.ConclusionThis umbrella review suggests that natural products and dietary interventions have beneficial therapeutic effects on liver enzymes levels. Further clinical trials are necessary to establish the effectiveness of supplements that reduce liver enzymes.</p

Table_2_Natural products and dietary interventions on liver enzymes: an umbrella review and evidence map.DOCX

BackgroundThe association between natural products and dietary interventions on liver enzymes is unclear; therefore, this study aimed to examine their effects on liver enzymes in adults.MethodsPubMed, Embase, and Cochrane Library of Systematic Reviews databases were searched from inception until March 2023. The Assessment of Multiple Systematic Reviews-2 (AMSTAR-2) and Grading of Recommendations Assessment, Development, and Evaluation (GRADE) systems were used to assess the methodological and evidence quality, and the therapeutic effects were summarized in a narrative form.ResultsA total of 40 meta-analyses on natural products (n = 25), dietary supplements (n = 10), and dietary patterns (n = 5) were evaluated, and results were presented in a narrative form. The overall methodological quality of the included studies was relatively poor. The results indicated that positive effects were observed for nigella sativa, garlic, artichoke, curcumin, silymarin, vitamin E, vitamin D, L-carnitine, propolis, and polyunsaturated fatty acids on certain liver enzymes. The dietary patterns, including high-protein, Mediterranean, and calorie-restriction diets and evening snacks, may reduce liver enzymes; however, other supplements and herbs did not reduce liver enzyme levels or have minimal effects. The evidence quality was generally weak given the risk of bias, heterogeneity, and imprecision.ConclusionThis umbrella review suggests that natural products and dietary interventions have beneficial therapeutic effects on liver enzymes levels. Further clinical trials are necessary to establish the effectiveness of supplements that reduce liver enzymes.</p