Journal: The EMBO Journal

Article Title: Arthropod exosomal glycine-rich protein as a potential vaccine candidate effectively reduces tick blood-feeding and pathogen transmission

doi: 10.1038/s44318-026-00709-z

Figure Lengend Snippet: Hematoxylin and eosin (H&E) staining of skin biopsy samples from mice (that allowed feeding of mock/ XM_002400035 -dsRNA-treated ticks) display that tick feeding causes inflammation at bite site in mock-dsRNA-treated group ( A ), but inflammation is reduced in XM_002400035 -dsRNA-treated group ( B ). Within the panniculus, there is downward projection of epidermis containing chitinous tick mouthparts (shown by black arrow). The panniculus contained moderate to large number of neutrophils, lymphocytes, plasma cells, and lower number of macrophages in mice that allowed feeding of mock-dsRNA-treated ticks ( A ). However, panniculus contained moderate number of inflammatory cells (shown by black arrow) with lymphocytes mixed with few macrophages and plasma cells in mice that allowed feeding of XM_002400035 -dsRNA-treated ticks. There is a mild crush artifact in this image. Magnification of both these images is 200×. Scale bar indicates 100 μm for each image. Enlarged images shown in Fig. 9A,B are repeated in Appendix Fig. for better visualization. ( C ) ELISA assay performed with skin lysates from mice that allowed feeding of ticks silenced for exosomal GRP or mock control ticks. Samples were probed with serum from immunized mice (1:1000 dilution). ( D ) Scratch assays performed on HaCaT cell monolayers incubated with 2 µg of GST/GST- GRP/GST-CXCL-12 protein (for 12 h), with/without 20 µl of tick exosomes from uninfected (UI), LGTV-infected (I), LGTV-infected and mock-dsRNA-treated or LGTV-infected and XM_002400035 -dsRNA-treated groups are shown. Phase contrast images (obtained using EVOS auto-fluorescence system, M7000) of HaCaT cell monolayers were taken for selected time-points (as before scratch, 0, 16, 20, and 24 h) and using 10× magnification. Untreated (UT) monolayers served as internal control. Scale bar indicates 275 μm for each image per group or timepoint. ( E ) Measurement of remaining wound size diameters (analyzed by ImageJ software) at different time-points (of 0, 16, 20, and 24 h) post-treatment of tick exosomes-derived from UI, I, mock/ XM_002400035 -dsRNA is shown. Wounds at 0 h were considered as 100% for all groups, including untreated (UT) control. Mouse CXCL-12 expression was analyzed in skin samples from mice immunized with GST/GST-GRP protein is shown ( F ). Exact number of sample numbers for each group representing multiple experiments is 5 mice for GST/6 mice for GST-GRP groups (in C , F ). Statistical differences were calculated using Mann–Whitney U test and p value is shown. p < 0.05 is considered as statistically significant. ( G ) Schematic model showing tick-borne flavivirus transmission to vertebrate host via tick saliva-derived exosomes. Ixodes scapularis tick attaches firmly and bites on host skin for longer feeding. Secreted saliva contains a plethora of substances including cement and perhaps cement-like GRPs to seal the feeding cone/cavity for directional blood flow and to defend from being groomed off by the vertebrate host. During blood meal ingestion, infected-ticks may continuously spit saliva containing infectious exosomes with viral full-length RNA genomes or polyproteins at host skin interface. We propose that incubation of tick exosomes containing exosomal GRP modulates the battle ground at skin interface by delaying cell migration/recruitment of immune cells (like neutrophils and dendritic cells from circulation) at the wound/bite site. Tick exosomes containing GRP inhibits residential keratinocytes and IL-8/CXCL-12 to delay injury, wound-healing, tissue damage, and repair process that will eventually enable ticks to acquire a successful blood meal at the host skin interface. .

Article Snippet: Keratinocyte growth kit , ATCC , # PCS-200-040.

Techniques: Staining, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Incubation, Infection, Fluorescence, Software, Derivative Assay, Expressing, MANN-WHITNEY, Transmission Assay, Migration

Journal: Gene therapy

Article Title: Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation.

doi: 10.1038/gt.2008.77

Figure Lengend Snippet: Figure 3 Mouse UL16-binding protein-like transcript 1 E (MULT1E)/FasTI induces apoptosis. A total of 1 106 cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml NKG2D/Fc for 16 h. The cells were then analyzed for apoptosis and necrosis using Annexin V assay (a–c) or caspase-3 assay (d) according to the manufacturers’ protocols. (a) An example of the fluorescence-activated cell sorting (FACS) data. (b, c) Summaries of data from three separate experiments. The statistical analyses were conducted between the controls (open bars) and NKG2D/Fc-treated cells (solid bars) using two-way analysis of variance (ANOVA). The difference between NKG2D/Fc-treated L-5 cells and NKG2D/Fc-treated L-10 cells was also compared using Student’s t-test. *Po0.05; **Po0.01 and ***Po0.001.

Article Snippet: Induction of apoptosis in cells expressing the fusion protein To determine if cells expressing the fusion protein can be induced to undergo apoptosis, one million cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml of NKG2D/Fc for 16 h. Apoptosis of the cells was measured using two systems: a TACS Annexin V-FITC Apoptosis Kit (R&D Systems) and a caspase-3 fluorometric assay (R&D Systems).

Techniques: Binding Assay, Clone Assay, Annexin V Assay, Caspase-3 Assay, FACS

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: Elevated soluble CD14 (sCD14) concentration in the serum of cirrhotic patients. Concentration of sCD14 was measured in healthy controls (n=31), patients with chronic viral hepatitis (n=26) and patients with cirrhosis (n=50) as described in patients and methods. Data is presented graphically as Whisker box-plots

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques: Concentration Assay, Whisker Assay

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: High correlation between the levels of human beta defensin-1 (hBD-1) and soluble CD14 (sCD14) in the hepatic veins of cirrhotic patients . Concentrations of hBD-1 and sCD14 were measured as described in patients and methods. Each dot corresponds to individual patients with cirrhosis (n=45). Analysis was performed in samples collected from peripheral veins (n=25, ) and from hepatic veins (n=20, )

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques:

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: Soluble CD14 (sCD14) and lipopolysaccharide binding protein (LBP) strongly correlate in serum of patients with cirrhosis. Concentrations of sCD14 were measured as described in Patients and Methods. Concentrations of LBP were measured by a commercially available ELISA, according to manufacturer’s instructions. Each dot corresponds to individual patients with cirrhosis (n=36). Analysis was performed in samples collected from peripheral veins

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Human Platelet Lysates‐Based Hydrogels: A Novel Personalized 3D Platform for Spheroid Invasion Assessment

doi: 10.1002/advs.201902398

Figure Lengend Snippet: Protein and growth factor release from PLMA hydrogels. A) Total protein quantification and B,C) ELISA quantification of TGF‐β1 and VEGF‐A release from PLMA hydrogels at 10, 15, and 20% (w/v). Data are presented as mean ± SD ( n ≥ 3).

Article Snippet: ELISA assays were performed to quantify the release of vascular endothelial growth factor (VEGF Human ELISA Kit, Invitrogen, ThermoFisher Scientific, USA), transforming growth factor β1 (TGF‐β1 Human ELISA Kit, Invitrogen, ThermoFisher Scientific, USA), and epidermal growth factor (Human EGF Quantikine ELISA Kit, R&D systems, Minneapolis, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of food and drug analysis

Article Title: Suppression of ERK1/2 and hypoxia pathways by four Phyllanthus species inhibits metastasis of human breast cancer cells.

doi: 10.1016/j.jfda.2016.03.010

Figure Lengend Snippet: Figure 2 e Protein-expression levels in (A) untreated cells and (B) representative images of cells treated with aqueous Phyllanthus watsonii. (C) Western blot showing VEGF expression in untreated cells and cells treated with extracts from the four Phyllanthus species. (d) Percentage of individual protein expression. (E, F) MMP expression in cells after treatment with aqueous and methanolic extracts from four Phyllanthus species, respectively. ELISA-based detection of (G) iNOS and (H) VEGF expression in untreated cells treated with extracts from the four Phyllanthus species. Error bars indicate the standard error of the mean of three independent experiments. p < 0.05 for each Phyllanthus treatment as compared with the untreated group. APN ¼ aqueous P. niruri; APU ¼ aqueous P. urinaria; APW ¼ aqueous P. watsonii; APA ¼ aqueous P. amarus; C ¼ untreated control; Cis ¼ cisplatin; Dox ¼ doxorubicin; ELISA ¼ enzyme-linked immunosorbent assay; H ¼ 500 mg/mL; I ¼ IC50 dosage; iNOS ¼ inducible nitric oxide synthase; L ¼ 50 mg/mL; M ¼ DNA marker; MMP ¼ matrix metalloproteinase; MPA ¼ methanolic P. amarus; MPN ¼ methanolic P. niruri; MPU ¼ methanolic P. urinaria; MPW ¼ methanolic P. watsonii; VEGF ¼ vascular endothelial growth factor.

Article Snippet: Total iNOS in the four Phyllanthus species-treated cells was measured using a human total-iNOS immunoassay enzymelinked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN, USA) according to manufacturer instructions.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Marker

Journal: Pediatric Reports

Article Title: Resistin in Urine and Breast Milk: Relation to Type of Feeding and Anthropometry at 1-Month

doi: 10.3390/pediatric14010013

Figure Lengend Snippet: Correlation between resistin concentration in the breast milk and ( a ) body weight; ( b ) age; ( c ) body mass index (BMI); and ( d ) leptin concentration in mothers’ breast milk at 1 month post-partum.

Article Snippet: We analyzed urinary and breast milk resistin levels (Human Resistin ELISA kit; Arigo biolaboratories, Hsinchu City, Taiwan) and breastmilk leptin (Human Leptin Quantikine ELISA Kit; R&D Systems, Minneapolis, MI, USA) in infants using an enzyme-linked immunosorbent assay kit.

Techniques: Concentration Assay