Journal: Clinical Cancer Research

Article Title: Anti-EGFR Antibody–Drug Conjugate Carrying an Inhibitor Targeting CDK Restricts Triple-Negative Breast Cancer Growth

doi: 10.1158/1078-0432.CCR-23-3110

Figure Lengend Snippet: Stochastic conjugation of cetuximab to CDK inhibitor and ADC internalization in live breast cancer cells. A, Flow cytometric evaluation of surface EGFR expression (TNBC: MDA-MB-468, HCC1143, HCC1806, MDA-MB-231, HCC1937, SUM149, and CAL51; HER2+: SKBR3; ER+: MCF7, T47D; nontumorigenic epithelial cell model: MCF10A; immune cell model: human B lymphocytes RPMI8866, RPMI8226, and monocytic cell line U937; human primary melanocyte: melanocyte). B, EGFR mRNA expression from the Cancer Cell Line Encyclopedia database showed a positive correlation with surface EGFR measured by flow cytometry in A (Spearman’s rank coefficient, r = 0.723). A high level of correlation was found between EGFR and cyclin E ( r = 0.738), but not with cyclin A or CDK2. Nonsignificant P values are marked as NS. C, Top, Schematic diagram of stochastic ADC conjugation by antibody reduction with TCEP and then conjugation to SNS-032 via MC-Val–Ala-PAB. Middle, HIC analysis confirmed an average DAR of 4.4. Bottom, SEC trace indicates negligible ADC aggregation and minimal free linker–payload (less than 0.8%). D, Surface plasmon resonance analysis demonstrated similar binding affinity ( K D ) for cetuximab (0.73 nmol/L) and ADC (1.28 nmol/L). Isotype IgG1 and isotype ADC showed no measurable binding. E, Monitoring internalization of Fabfluor-pH-labeled cetuximab, ADC, or isotype control (10 nmol/L) by Incucyte live-cell imaging. Phase and red fluorescence time-course images were captured for 24 hours. Images of internalized antibody display in cytosolic, low pH lysosomal vesicle-associated red fluorescence in cells. Scale bar, 0.2 mm. F, Cells were seeded in Matrigel for 5 days, allowing the formation of spheroids. Fabfluor-pH-labeled antibodies or ADC (10 nmol/L) were introduced in the Matrigel and showed rapid internalization in EGFR-high MDA-MB-468 and MDA-MB-231, whereas EGFR-low CAL51 displayed little red fluorescence signals. A low level of internalization was observed for isotype or isotype-ADC controls. Scale bar, 0.5 mm. P values determined by two-tailed unpaired t test of three independent experiments compared with isotype control.

Article Snippet: Human primary epidermal melanocytes were cultured in Melanocyte Growth Medium (Cell Applications Inc.).

Techniques: Conjugation Assay, Expressing, Flow Cytometry, SPR Assay, Binding Assay, Labeling, Control, Live Cell Imaging, Fluorescence, Two Tailed Test

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: Effects of TPS on the expression of α-MSH. Representative chart trace of α-MSH inhibition by 5, 10, and 15 μg/ml of the TPS on HaCaT cells ( A ) and HEM cells ( B ) with UVA exposure (15 J/cm 2 ) or not by ELISA. Error bars show means ± SEMs. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: TPS inhibited melanogenesis through suppressing the α-MSH-MC1R signalling pathway in HaCaT cells and HEM cells. A, B – HEM was pretreated or not with UVA exposure (15 J/cm 2 ) before TPS (10 μg/ml) and N-1A (20 μm) were applied. Melanin contents and the expression of tyrosinase were measured as described in methods. C – HaCaT cells were treated with TPS (10 μg/ml) and/or N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. D – HEM cells were treated with TPS (10 μg/ml) and N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells, a p < 0.05, aa p < 0.01, aaa p < 0.001, ns p > 0.05

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Nutrients

Article Title: Evaluation of the Relationship Between Orexin A, Peptide YY, AgRP, and POMC Levels and Sleep Disorders in Children with Malnutrition

doi: 10.3390/nu18030377

Figure Lengend Snippet: Box plots showing the distribution of the orexin A, proopiomelanocortin (POMC), agouti-related protein (AgRP), and peptide yy (PYY) levels in the malnourished children and typically developing (TD) healthy controls. The Mann–Whitney U test was used to compare the peptide levels between the two groups.

Article Snippet: Human POMC, orexin A, AgRP, and PYY enzyme-linked immunosorbent assay (ELISA) kits (Shanghai Korain Biotech Co., Ltd., Shanghai, China) were used to measure the plasma peptide levels.

Techniques: MANN-WHITNEY