Journal: Diagnostics

Article Title: Phonosurgical Treatment of Laryngeal Leukoplakia and Dysplasia: Results of Multidimensional Voice Diagnostics Including the VEM

doi: 10.3390/diagnostics16081242

Figure Lengend Snippet: Intraoperative comparison of treatment modalities: Removal of leukoplakic vocal fold changes using cold steel forceps ( left ), CO 2 laser ( center ), and blue laser ( right ).

Article Snippet: This involved the use of laryngeal scissors and forceps (Storz, Tuttlingen, Germany), the AcuPulse 30W/40 ST CO 2 laser system (Lumenis, Yokneam, Israel), and the WOLF TrueBlue surgical laser (A.R.C.

Techniques: Comparison

Journal: The Journal of Biological Chemistry

Article Title: Tryptophan metabolites 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) increase oxidative stress and impair osteoblastic bone formation

doi: 10.1016/j.jbc.2026.111174

Figure Lengend Snippet: Tryptophan and its downstream metabolites activate AhR transcriptional activity . A , luciferase assay results showing activation of AhR transcriptional activity by tryptophan and its downstream metabolites. AhR transcriptional activity after inhibiting IDO1 with PF-06840003 (10 μM) ( B ) and Kyn-3-monooxygenase with JM6 (5 μM) ( C ) and treating ST2 cells with tryptophan and KYN respectively. D , Time course qPCR experiment showing Cyp1a1 expression in C2C12 cells following KYN treatment. C2C12 cells were treated with tryptophan metabolites for 3 h ( E ) and 24 h ( F ) and the mRNA levels of AhR target genes were measured via RT-qPCR. Multiple group comparison was performed by one way ANOVA and Fisher’s least significant difference (LSD) post hoc testing and bars with different superscript letters are significantly ( p < 0.05) different from one another (n ≥ 3 biological replicates for each condition). Cyp1a1, cytochrome p450 1a1; Cyp1b1, cytochrome p450 1b1; Ahrr, Aryl hydrocarbon repressor. # indicates significant difference versus vehicle control at p < 0.05 (#), p < 0.01 (##), p < 0.001 (###) as shown by one sample t-tests versus vehicle controls. AhR, aryl hydrocarbon receptor; Kyn, kynurenine.

Article Snippet: Immortalized murine ST2 (obtained from Leibniz Institute DSMZ, #ACC333) and C2C12 (obtained from ATCC, #CRL-1772) cell lines were used for in vitro studies.

Techniques: Activity Assay, Luciferase, Activation Assay, Expressing, Quantitative RT-PCR, Comparison, Control

Journal: The Journal of Biological Chemistry

Article Title: Tryptophan metabolites 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) increase oxidative stress and impair osteoblastic bone formation

doi: 10.1016/j.jbc.2026.111174

Figure Lengend Snippet: 3-hydroxy-kynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) induce DNA damage leading to senescence at low concentrations and apoptotic cell death at higher concentration . DNA damage was evaluated after treating ST2 cells with 3HK ( A ), 3HAA ( B ), Tryptophan (Tryp) ( C ) and Kyn ( D ) then staining the cells with Gamma H2AX antibody. ( E ) ST2 cells were treated with tryptophan metabolites (100 μM) or doxorubicin as a positive control (1 μM) for 24 h, fixed and stained for β-galactosidase activity to reflect senescence, β-galactosidase positive cells were quantified using the manual cell counting function of ImageJ. MTT assays were used to measure cell viability after treating ( F ) C2C12 and ( G ) ST2 cells with tryptophan metabolites for 24 h. H , representative images showing that 3HK (400 μM) and 3HAA (300 μM) induced apoptosis in C2C12 cells as indicated by positive TUNEL staining ( left ); ImageJ was utilized to quantify TUNEL labeling from n ≥ 3 biological replicates ( right ), and the results were analyzed by Student’s t test. The scale bar represents 50 μm. MTT assay showing that scavenging ROS with N-Acetylcysteine rescued ( I ) 3HK and ( J ) 3HAA-induced cell death. Multiple group comparison was performed by one way ANOVA and Fisher’s LSD post hoc testing and bars with different superscript letters are significantly ( p < 0.05) different from one another (n ≥ 3 biological replicates for each condition). 3HAA, 3-hydroxyanthranilic acid; 3HK, 3-hydroxy-kynurenine.

Article Snippet: Immortalized murine ST2 (obtained from Leibniz Institute DSMZ, #ACC333) and C2C12 (obtained from ATCC, #CRL-1772) cell lines were used for in vitro studies.

Techniques: Concentration Assay, Staining, Positive Control, Activity Assay, Cell Counting, TUNEL Assay, Labeling, MTT Assay, Comparison

Journal: The Journal of Biological Chemistry

Article Title: Tryptophan metabolites 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) increase oxidative stress and impair osteoblastic bone formation

doi: 10.1016/j.jbc.2026.111174

Figure Lengend Snippet: HK and 3HAA induce oxidative stress in mesenchymal lineage cells . Amplex red assay measuring whole cell ROS production after 6 h of treatment with ( A and C ) 3HK and ( B and D ) 3HAA in the presence and absence of the ROS scavenger N-acetyl cysteine (5 mM). One sample t test was used to compare treatment conditions with their vehicle control # indicates significant difference versus veh. at p < 0.05 (#), p < 0.01 (##), p < 0.001 (###). Amplex red assay was used to measure mitochondrial ROS ( E ) while tetramethylrhodamine methyl ester measured transmembrane potential ( F ) after 6 h treatment of ST2 cells with tryptophan metabolites. Multiple group comparison was done by one way ANOVA and Fisher’s LSD post hoc testing and bars with different superscript letters are significantly ( p < 0.05) different from one another (n ≥ 3 biological replicates for each condition). The expression of the antioxidant enzyme Hmox1 and ( G and H ) its transcription factor regulator Nrf-2 ( I and J ) was increased following a 24 h treatment of ST2 cells with 3HK and 3HAA. One sample t test was used to compare treatment conditions with their vehicle control; # indicates significant difference versus veh. at p < 0.05 (#), p < 0.01 (##), p < 0.001 (###). 3HAA, 3-hydroxyanthranilic acid; 3HK, 3-hydroxy-kynurenine; ROS, reactive oxygen species.

Article Snippet: Immortalized murine ST2 (obtained from Leibniz Institute DSMZ, #ACC333) and C2C12 (obtained from ATCC, #CRL-1772) cell lines were used for in vitro studies.

Techniques: Amplex Red Assay, Control, Comparison, Expressing

Journal: bioRxiv

Article Title: Bacterial exo-α-sialidases subvert the complement system through desialylation

doi: 10.64898/2026.02.05.703967

Figure Lengend Snippet: (a) SDS-PAGE analysis of six purified recombinant sialidases, including Td -NanH, Cp -NanI, Cp -NanH, Bf -NanH2, Tf -NanH, and Vc -NanH. Cp -NanI was purchased from Sigma-Aldrich, N2133 and the remaining five sialidases were expressed in E. coli and purified using FPLC. (b) Kinetic analysis of six GH33 sialidases. This assay was carried out as described in the Methods section using 4-methylumbelliferyl-α-D-N-acetylneuraminic acid (4-MUNANA) as a substrate. Saturation curves were fitted to Michaelis-Menten kinetics using GraphPad Prism ; and K m and V max were calculated (see details in Supplementary Table 5). (c-e) GH33 sialidases desialylate human serum, C4 and FH proteins. For this study, normal human serum (NHS), C4 or FH proteins were incubated with six recombinant sialidases at 37°C for 1 or 3 hours, followed by SDS-PAGE and lectin blots using SNA. Abbreviations: Td, Td- NanH; Cp-I, Cp- NanI; Cp-H, Cp- NanH; Bf, Bf- NanH2; Tf, Tf- NanH; and Vc, Vc- NanH.

Article Snippet: Briefly, 50 nM purified bacterial sialidases were incubated with increasing concentrations (15.625 - 1250 μM) of either 2′-(4-methylumbelliferyl)-α-D- N -acetylneuraminic acid (4-MUNANA; GoldBio, St. Louis, MO) or 4-methylumbelliferyl α- N -glycolylneuraminic acid (4-MUNGNA; Sussex Research, Ontario, Canada) in the appropriate reaction buffer.

Techniques: SDS Page, Purification, Recombinant, Incubation

Journal: bioRxiv

Article Title: Bacterial exo-α-sialidases subvert the complement system through desialylation

doi: 10.64898/2026.02.05.703967

Figure Lengend Snippet: (a) Schematic illustration of the domain architecture of T. denticola NanH ( Td -NanH). Annotated features include the signal peptide, the conserved “RIP” motif, and three Asp-box motifs with their respective amino acid positions. InlB: Listeria monocytogenes Internalin B. (b-c) Enzymatic kinetics of Td -NanH toward the fluorogenic substrates 4-MUNANA and 4-MUNAGc. Assays were performed as described in using wild-type Td -NanH and three point mutants including Td -NanH R140A , Td -NanH P142A , and Td -NanH R140AP142A ; and as C-terminal fragment of Td -NanH ( Td-C ) (d) Serum bactericidal assays. T. denticola wild-type, the Td -NanH deletion mutant (Δ Tde471 ), and the isogenic complemented strain ( CTde471 ) were incubated with 25% normal human serum (NHS) or heat-inactivated serum (HIS) for 30 min at 37 °C. Viable bacteria were quantified using a Petroff– Hausser counting chamber. Survival was calculated as the ratio of viable cells in NHS relative to HIS. Data are presented as mean ± SEM and analyzed by one-way ANOVA with Tukey’s multiple comparisons test ( P < 0 . 01 ). (e) Td -NanH restores serum resistance in the Δ Tde471 mutant. NHS (25%) was pre-treated with recombinant Td -NanH or the catalytically inactive mutant Td -NanH * for 1-3 h at 37 °C prior to serum killing assays with the Δ Tde471 strain using the same protocol described in (d). (f) GH33 sialidases broadly inhibit serum bactericidal activity. NHS (25%) was pre-treated with six GH33 sialidases for 1 h at 37 °C and subsequently used in serum killing assays with the Δ Tde471 mutant, following the protocol in (d). (g) Complement deposition on Δ Tde471 cells. A total of 5 × 10□ Δ Tde471 cells were incubated with 25% NHS for 15 min at 37 °C under three conditions: untreated NHS, Td -NanH-treated NHS, or Td -NanH * -treated NHS. Cell-associated complement components were analyzed by SDS-PAGE and immunoblotting using antibodies against C3 and C9. A T. denticola specific antibody served as a loading control.

Article Snippet: Briefly, 50 nM purified bacterial sialidases were incubated with increasing concentrations (15.625 - 1250 μM) of either 2′-(4-methylumbelliferyl)-α-D- N -acetylneuraminic acid (4-MUNANA; GoldBio, St. Louis, MO) or 4-methylumbelliferyl α- N -glycolylneuraminic acid (4-MUNGNA; Sussex Research, Ontario, Canada) in the appropriate reaction buffer.

Techniques: Mutagenesis, Incubation, Bacteria, Recombinant, Activity Assay, SDS Page, Western Blot, Control