Journal: Journal of Clinical Medicine

Article Title: Blood Biomarkers for Alzheimer’s Disease in Down Syndrome

doi: 10.3390/jcm10163639

Figure Lengend Snippet: Blood biomarkers for non ATN-processes proposed for AD dementia in DS. Biomarkers are grouped according to the molecular pathway pathologically affected in DS. Colors for each group of biomarkers are related to Figure 2.

Article Snippet: , MMP-1 , , Mesoscale , Multi-Spot MMP 3-Plex Ultra-Sensitive kit SECTOR Imager 2400 (MesoScale Discovery) , DS > controls DS-AD > controls.

Techniques: Biomarker Discovery, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Western Blot, Targeted Proteomics, Spectrophotometry, Activity Assay, Methylation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An exhaustive cell-based screen coupled with an intracellular-induced lux-based reporter identified bioactive molecules that inhibit host cell infection by intracellular pathogens

doi: 10.3389/fcimb.2026.1770677

Figure Lengend Snippet: The effect of the identified compounds on host cell invasion and intracellular replication in epithelial cells. HeLa cells were seeded in a 24-well cell culture-treated microplate and incubated with fresh DMEM medium containing compounds C1 to C8 at a final concentration of 10 µM for 3 h prior to the infection. Cells were infected at an MOI of 50, incubated at 37°C under 5% CO 2 , and then subjected to the gentamicin protection assay as explained in the Materials and methods section to eliminate extracellular bacteria. At 2 hpi, cells were washed and lysed with lysis buffer. To calculate the number of invading bacteria at 2 hpi (A) , the cell lysates were serially diluted in PBS and plated onto LB-agar plates for CFU count. An S . Typhimurium invA null mutant strain impaired in invasion of non-phagocytic cells was included as a genetic control. To calculate the intracellular fold replication (B) , at 2 hpi, the medium of the infected cells was replaced with DMEM supplemented 10 µM of each compound and 10 µg/mL of gentamicin and incubated at 37°C in a humidified 5% CO 2 incubator. At 24 h post-infection, the cells were washed and lysed as above. Intracellular replication was determined by the ratio between the number of intracellular cells counted at 24 hpi and their number at 2 hpi. The percentage of invasion and replication relative to HeLa cells infected with S . Typhimurium in the absence of the compounds (Ctrl) is shown. HT29-MTX-E12 cells were incubated with the indicated inhibitors for 3 h and infected with a mid-logarithmic culture of S. Typhimurium SL1344 at an MOI of 30. Infected cells were subjected to the gentamicin protection assay. (C) At 2 hpi, Salmonella invasion into HT29-MTX-E12 cells was determined as in (A) . (D) At 24 h after infection, cells were washed and lysed with lysis buffer. Serial dilutions were plated on LB agar plates for CFU counting, and intracellular replication was determined as in (B) . The charts show the mean and standard error of the mean (SEM) of at least three biological repeats. One-way ANOVA was used to determine statistical significance. *, P-value <0.05; **, P-value <0.01; ***, P-value < 0.001; ns, not statistically significant.

Article Snippet: Absorbance at 600 nm and bioluminescence were determined using the Infinite 200 PRO M-Plex microplate reader (Tecan, Männedorf, Switzerland) and imaged using IVIS Lumina LT (PerkinElmer, Shelton, USA) after 5.5 h of growth.

Techniques: Cell Culture, Incubation, Concentration Assay, Infection, Bacteria, Lysis, Mutagenesis, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An exhaustive cell-based screen coupled with an intracellular-induced lux-based reporter identified bioactive molecules that inhibit host cell infection by intracellular pathogens

doi: 10.3389/fcimb.2026.1770677

Figure Lengend Snippet: Structure–activity relationship (SAR) studies of compound C4. (A) HeLa cells were seeded in a 96-well, cell culture-treated, microplate and incubated at 37°C in 5% CO 2 humidified incubator for 4 h in the presence of 10 µM of compound C4 (PCM-0103431) and its chemical analogs (shown as gray bars). As a positive control, 2 µM of cytochalasin D (CD) was also included. Cells were infected with subcultures of S . Typhimurium expressing the reporter system P ssek3::lux at an MOI of 50. One hpi, gentamicin was added to a final concentration of 20 µg/mL, and the infected cells were incubated overnight, until bioluminescence was read. Host cell infection is shown as the intracellular luminescence relative to its value in HeLa cells infected with S. Typhimurium in the presence of 0.1% DMSO (Ctrl). The charts show the mean and SEM of three biological repeats. One-way ANOVA was used to determine statistical significance. (B) SAR diagram of compound 4 (PCM-0103431) of various modifications made to the pyran scaffold. In green, the aniline moiety was substituted with electron-donating and neutral groups. In red, changes to the amine were not tolerated; a secondary amine was crucial for activity. Magenta: methyl groups could be replaced with ethyl, maintaining activity. Blue: p-methoxypheny, tolyl, and benzyl groups maintained their activity. **, P-value <0.01; ***, P-value < 0.001; ns, not statistically significant.

Article Snippet: Absorbance at 600 nm and bioluminescence were determined using the Infinite 200 PRO M-Plex microplate reader (Tecan, Männedorf, Switzerland) and imaged using IVIS Lumina LT (PerkinElmer, Shelton, USA) after 5.5 h of growth.

Techniques: Activity Assay, Cell Culture, Incubation, Positive Control, Infection, Expressing, Concentration Assay