Journal: Scientific Reports

Article Title: Fluorescence-based reagent and spectrum-based optical reader for lactoferrin detection in tears: differentiating Sjögren’s syndrome from non-Sjögren’s dry eye syndrome

doi: 10.1038/s41598-024-65487-2

Figure Lengend Snippet: The principle and demonstration of the photo-detection device are shown. ( A ) The Tb 3+ –lactoferrin complex gets excited by ultraviolet light at 290 nm. The energy is transferred through tyrosine at the binding site, and fluorescein-emitted light at a wavelength of 570 nm is obtained, detected by optical fiber and amplified by wafer technology. ( B ) Samples are added to the cuvette. ( C ) Fluorescence levels are checked using the corresponding APP. ( D ) The photo-detection device (yellow arrow), cuvette (white arrow), and its APP are shown. ( E ) Different fluorescence intensities under different concentrations of lactoferrin are shown. APP application.

Article Snippet: Standard lactoferrin level references were established by detecting the fluorescence intensities of different concentrations of diluted lactoferrin from human milk (L0520, Sigma-Aldrich, Darmstadt, Germany) (Fig. E).

Techniques: Binding Assay, Amplification, Fluorescence

Journal: Scientific Reports

Article Title: Fluorescence-based reagent and spectrum-based optical reader for lactoferrin detection in tears: differentiating Sjögren’s syndrome from non-Sjögren’s dry eye syndrome

doi: 10.1038/s41598-024-65487-2

Figure Lengend Snippet: Representative pictures of the outer segment of eyes from patients with non-Sjögren’s DED ( A , C ) and SS-DED ( B , D ) are shown. ( A ) Picture from a patient with non-Sjögren’s DED, with a Schirmer test result of 4 mm, OSDI of 32, and tear lactoferrin level of 0.49 mg/mL, is shown. ( B ) Picture from a patient with SS-DED, with a Schirmer test result of 1 mm, OSDI of 33, and tear lactoferrin level of 0.05 mg/mL, is shown. ( C ) The fluorescein staining result of the patient in ( A ) is shown, with no obvious superficial punctate keratitis. ( D ) The fluorescein staining picture of the patient in ( B ) is shown, revealing diffuse corneal erosions (arrow). DED dry eye disease, OSDI Ocular Surface Disease Index, SS-DED Sjögren’s syndrome DED.

Article Snippet: Standard lactoferrin level references were established by detecting the fluorescence intensities of different concentrations of diluted lactoferrin from human milk (L0520, Sigma-Aldrich, Darmstadt, Germany) (Fig. E).

Techniques: Staining

Journal: Scientific Reports

Article Title: Fluorescence-based reagent and spectrum-based optical reader for lactoferrin detection in tears: differentiating Sjögren’s syndrome from non-Sjögren’s dry eye syndrome

doi: 10.1038/s41598-024-65487-2

Figure Lengend Snippet: The correlation between the lactoferrin (LF) concentration as assessed using the photo-detection device and with ELISA is depicted. The concentration of LF as assessed using the photo-detection device is highly correlated to that as assessed using ELISA. R 2 = 0.9724. X-axis: the results from ELISA; Y- axis: the results from the photo-detection device. ELISA enzyme-linked immunosorbent assay, LF lactoferrin.

Article Snippet: Standard lactoferrin level references were established by detecting the fluorescence intensities of different concentrations of diluted lactoferrin from human milk (L0520, Sigma-Aldrich, Darmstadt, Germany) (Fig. E).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Fluorescence-based reagent and spectrum-based optical reader for lactoferrin detection in tears: differentiating Sjögren’s syndrome from non-Sjögren’s dry eye syndrome

doi: 10.1038/s41598-024-65487-2

Figure Lengend Snippet: Comparisons in tear lactoferrin levels between non-Sjögren’s DED, SS-DED, and control groups are shown. Patients with non-Sjögren’s DED and SS-DED show significantly lower levels of tear lactoferrin than healthy controls (p < 0.0001). Further, patients with SS-DED show lower tear lactoferrin levels than patients with non-Sjögren’s DED (p < 0.001). DED dry eye disease, SS-DED Sjögren’s syndrome DED.

Article Snippet: Standard lactoferrin level references were established by detecting the fluorescence intensities of different concentrations of diluted lactoferrin from human milk (L0520, Sigma-Aldrich, Darmstadt, Germany) (Fig. E).

Techniques: Control

Journal: Scientific Reports

Article Title: Fluorescence-based reagent and spectrum-based optical reader for lactoferrin detection in tears: differentiating Sjögren’s syndrome from non-Sjögren’s dry eye syndrome

doi: 10.1038/s41598-024-65487-2

Figure Lengend Snippet: Association between tear lactoferrin level and other factors.

Article Snippet: Standard lactoferrin level references were established by detecting the fluorescence intensities of different concentrations of diluted lactoferrin from human milk (L0520, Sigma-Aldrich, Darmstadt, Germany) (Fig. E).

Techniques:

Journal: Virology Journal

Article Title: Human lactoferrin but not lysozyme neutralizes HSV-1 and inhibits HSV-1 replication and cell-to-cell spread

doi: 10.1186/1743-422X-6-53

Figure Lengend Snippet: Lactoferrin but not lysozyme inhibited efficiently HSV-1 infection of Vero cells . Replication of wt HSV-1 and mutant viruses gC - 39 and Rid1 in Vero cells with human non-iron-saturated Lf (apoLf), iron-saturated Lf (satLf) or lysozyme (hLz) present throughout the entire replication cycle. Lactoferrin and lysozyme were added to cell cultures 24 h prior to infection. The relative amount of viral plaques, in comparison to untreated cultures, is shown. Error bars represent standard deviations of three separate experiments. Statistical significance in the level of inhibition: * = p < 0.05, ** = p < 0.005, mutant viruses compared to wt KOS. Only results with the highest concentration of hLz used are shown.

Article Snippet: We infected Vero cell monolayers with HSV-1 in the presence or absence of human milk-derived non-iron-saturated lactoferrin (apoLf; L-0520, Sigma), iron-saturated lactoferrin (satLf; L-3770, Sigma) or lysozyme (hLz; L-6394, Sigma).

Techniques: Infection, Mutagenesis, Inhibition, Concentration Assay

Journal: Virology Journal

Article Title: Human lactoferrin but not lysozyme neutralizes HSV-1 and inhibits HSV-1 replication and cell-to-cell spread

doi: 10.1186/1743-422X-6-53

Figure Lengend Snippet: Lactoferrin but not lysozyme neutralized HSV-1 . Wt HSV-1 and mutant viruses gC - 39 and Rid1 were incubated for 30 min at 37°C with different concentrations of human non-iron-saturated Lf (apoLf), iron-saturated Lf (satLf) or lysozyme (hLz) prior to inoculation of Vero cell cultures. Preincubated virus/hLf mixtures were diluted 1:10 upon addition to the cell culture so that the inoculum contained 1–50 μg/ml of hLf. Therefore the observed inhibition could not have resulted from the hLf effect on the cells. The relative amount of viral plaques, in comparison to untreated cultures, is shown. Error bars represent standard deviations of three separate experiments. Statistical significance in the level of inhibition: * = p < 0.05, ** = p < 0.005, mutant viruses compared to wt KOS. Only results with the highest concentration of hLz used are shown.

Article Snippet: We infected Vero cell monolayers with HSV-1 in the presence or absence of human milk-derived non-iron-saturated lactoferrin (apoLf; L-0520, Sigma), iron-saturated lactoferrin (satLf; L-3770, Sigma) or lysozyme (hLz; L-6394, Sigma).

Techniques: Mutagenesis, Incubation, Cell Culture, Inhibition, Concentration Assay

Journal: Virology Journal

Article Title: Human lactoferrin but not lysozyme neutralizes HSV-1 and inhibits HSV-1 replication and cell-to-cell spread

doi: 10.1186/1743-422X-6-53

Figure Lengend Snippet: Pretreatment of cell cultures with lactoferrin but not lysozyme inhibited HSV-1 infection . Vero cell cultures were pretreated with human non-iron-saturated Lf (apoLf), iron-saturated Lf (satLf) or lysozyme (hLz) for 24 h prior to inoculation. To exclude lactoferrin and lysozyme effect on HSV-1 virion, these supplements were removed from the media by washing before inoculation of cultures with wt HSV-1 and mutant viruses gC - 39 and Rid1. The relative amount of viral plaques, in comparison to untreated cultures, is shown. Error bars represent standard deviations of three separate experiments. Statistical significance in the level of inhibition: * = p < 0.05, ** = p < 0.005, mutant viruses compared to wt KOS. Only results with the highest concentration of hLz used are shown.

Article Snippet: We infected Vero cell monolayers with HSV-1 in the presence or absence of human milk-derived non-iron-saturated lactoferrin (apoLf; L-0520, Sigma), iron-saturated lactoferrin (satLf; L-3770, Sigma) or lysozyme (hLz; L-6394, Sigma).

Techniques: Infection, Mutagenesis, Inhibition, Concentration Assay

Journal: Virology Journal

Article Title: Human lactoferrin but not lysozyme neutralizes HSV-1 and inhibits HSV-1 replication and cell-to-cell spread

doi: 10.1186/1743-422X-6-53

Figure Lengend Snippet: Lactoferrin has inhibitory activity against HSV-1 infection also after viral adsorption . To study the lactoferrin effect on post-attachment events, wt HSV-1 and mutant viruses gC - 39 and Rid1 were adsorbed to cells at 4°C, cultures were washed and human non-iron-saturated Lf (apoLf), iron-saturated Lf (satLf) or lysozyme (hLz) was added. The relative amount of viral plaques, in comparison to untreated cultures, is shown. Error bars represent standard deviations of three separate experiments. Statistical significance in the level of inhibition: * = p < 0.05, ** = p < 0.005, mutant viruses compared to wt KOS. Only results with the highest concentration of hLz used are shown.

Article Snippet: We infected Vero cell monolayers with HSV-1 in the presence or absence of human milk-derived non-iron-saturated lactoferrin (apoLf; L-0520, Sigma), iron-saturated lactoferrin (satLf; L-3770, Sigma) or lysozyme (hLz; L-6394, Sigma).

Techniques: Activity Assay, Infection, Adsorption, Mutagenesis, Inhibition, Concentration Assay

Journal: Virology Journal

Article Title: Human lactoferrin but not lysozyme neutralizes HSV-1 and inhibits HSV-1 replication and cell-to-cell spread

doi: 10.1186/1743-422X-6-53

Figure Lengend Snippet: Lactoferrin inhibition of HSV-1 cell-to-cell spread is mediated by gD . HSV-1 KOS (a, d), gC - 39 (b) and Rid1 (c) infection of Vero cells in the absence or presence of 500 μg/ml of human lactoferrin. Plaques formed by the infecting virus were visualized by crystal violet staining after 5 days of culture in the presence of human immunoglobulin (a-c) or by immunoperoxidase staining with polyclonal anti-HSV-1 antibody after 20 h of culture (d). Lactoferrin dramatically impaired the cell-to-cell spread of HSV-1 KOS and gC - 39, but not of Rid1.

Article Snippet: We infected Vero cell monolayers with HSV-1 in the presence or absence of human milk-derived non-iron-saturated lactoferrin (apoLf; L-0520, Sigma), iron-saturated lactoferrin (satLf; L-3770, Sigma) or lysozyme (hLz; L-6394, Sigma).

Techniques: Inhibition, Infection, Staining, Immunoperoxidase Staining

Journal: BMC Infectious Diseases

Article Title: Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae

doi: 10.1186/1471-2334-4-12

Figure Lengend Snippet: Radial inhibition assay results demonstrating the effect of innate immune molecules on otitis media pathogens. Representative radial inhibition assay for the measurement of the activity of innate immune molecules against NTHi 12, M. catarrhalis 035E (M cat), S. pneumoniae 3 (Sp 3) and S. pneumoniae 6B (Sp 6B). Panel A shows the results of testing three doses (50 μg, 100 μg and 200 μg) of human lysozyme (Lz) against the bacteria. Panel B shows the results of testing the lactoferrin (Lf) and the defensins against the bacteria. Three doses (4 μg, 10 μg and 40 μg) of human lactoferrin (Lf) were tested, as were two doses (4 μg and 10 μg) each of human β defensin-1 (hBD-1) and β defensin-2 (hBD-2). Each dose was delivered in a total volume of 4 μl. The control well received only solvent (0.01% acetic acid). The diameter of the inhibition zone was measured and averaged in three separate experiments.

Article Snippet: The antimicrobial peptides and proteins, human milk lysozyme (Sigma, St. Louis, MO), human milk apo-lactoferrin (0.03% iron content, Sigma, St. Louis, MO), human β defensin-1 and human β defensin-2 (Peptides International, Louisville, KY), were dissolved in 0.01% acetic acid/0.1% bovine serum albumin (Sigma, St. Louis, MO) and 4 μl was added to each well.

Techniques: Inhibition, Activity Assay

Journal: BMC Infectious Diseases

Article Title: Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae

doi: 10.1186/1471-2334-4-12

Figure Lengend Snippet: Measurements of the effect of human lysozyme (hLz), lactoferrin (hLf), β defensin-1 (HBD-1) and β defensin-2 (HBD-2) on the growth of otitis media pathogens, using the radial inhibition assay.

Article Snippet: The antimicrobial peptides and proteins, human milk lysozyme (Sigma, St. Louis, MO), human milk apo-lactoferrin (0.03% iron content, Sigma, St. Louis, MO), human β defensin-1 and human β defensin-2 (Peptides International, Louisville, KY), were dissolved in 0.01% acetic acid/0.1% bovine serum albumin (Sigma, St. Louis, MO) and 4 μl was added to each well.

Techniques: Inhibition

Journal: BMC Infectious Diseases

Article Title: Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae

doi: 10.1186/1471-2334-4-12

Figure Lengend Snippet: Measurements of the effect of 100 mM NaCl on the inhibition of the growth of otitis media pathogens by human lysozyme (hLz), lactoferrin (hLf), β defensin-1 (HBD-1) and β defensin-2 (HBD-2), using the radial inhibition assay.

Article Snippet: The antimicrobial peptides and proteins, human milk lysozyme (Sigma, St. Louis, MO), human milk apo-lactoferrin (0.03% iron content, Sigma, St. Louis, MO), human β defensin-1 and human β defensin-2 (Peptides International, Louisville, KY), were dissolved in 0.01% acetic acid/0.1% bovine serum albumin (Sigma, St. Louis, MO) and 4 μl was added to each well.

Techniques: Inhibition

Journal: PLoS ONE

Article Title: EndoE from Enterococcus faecalis Hydrolyzes the Glycans of the Biofilm Inhibiting Protein Lactoferrin and Mediates Growth

doi: 10.1371/journal.pone.0091035

Figure Lengend Snippet: A. Human lactoferrin (hLF) was incubated with EndoE, EndoE(E186Q) and EndoE(E662Q), separated on 10% SDS-PAGE and stained with Coomassie (upper panel), or electro-blotted onto PVDF membranes and was analyzed with ConA lectin (lower panel). Incubation of hLF with PBS was used as a negative control. B. Plasmon surface resonance assay to analyze binding of EndoE to hLF. The plots show binding of EndoE(E186Q) and EndoE(E662Q) to hLF.

Article Snippet: Activity of EndoE on human lactoferrin from milk (hLF) (Sigma-Aldrich) was tested as described with other glycoproteins previously .

Techniques: Incubation, SDS Page, Staining, Western Blot, Negative Control, Resonance Assay, Binding Assay

Journal: PLoS ONE

Article Title: EndoE from Enterococcus faecalis Hydrolyzes the Glycans of the Biofilm Inhibiting Protein Lactoferrin and Mediates Growth

doi: 10.1371/journal.pone.0091035

Figure Lengend Snippet: Biofilm formation of E. faecalis was measured using the crystal violet assay and is expressed as OD 550 . Human lactoferrin (hLF) was added either fully glycosylated (hLF) or deglycosylated (de-hLF) due to the treatment with EndoE. Error bars indicate the standard deviation from the mean of three independent experiments with three replicates. w/o: no hLF added.

Article Snippet: Activity of EndoE on human lactoferrin from milk (hLF) (Sigma-Aldrich) was tested as described with other glycoproteins previously .

Techniques: Crystal Violet Assay, Standard Deviation

Journal: PLoS ONE

Article Title: EndoE from Enterococcus faecalis Hydrolyzes the Glycans of the Biofilm Inhibiting Protein Lactoferrin and Mediates Growth

doi: 10.1371/journal.pone.0091035

Figure Lengend Snippet: A. Western blot analysis, using anti-lactoferrin antibodies, of human lactoferrin (hLF) bound to E. faecalis OG1. E. faecalis was incubated with indicated concentrations of hLF or EndoE treated hLF (de-hLF). E. faecalis cell extract was separated on 10% SDS-PAGE and electro-blotted onto PVDF membranes. Control: 1 µg hLF. B. Binding of different hLF concentrations to recombinant enolase immobilized to a microtiter plate. Anti-hLF antibodies were used to detect the binding of hLF to the enolase. Error bars indicate the standard deviation from the mean of three independent experiments.

Article Snippet: Activity of EndoE on human lactoferrin from milk (hLF) (Sigma-Aldrich) was tested as described with other glycoproteins previously .

Techniques: Western Blot, Incubation, SDS Page, Binding Assay, Recombinant, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Lactoferrin Inhibition of the Complex Formation between ACE2 Receptor and SARS CoV-2 Recognition Binding Domain

doi: 10.3390/ijms23105436

Figure Lengend Snippet: Time−courses of the reaction between RBD in solution with ACE2 in the absence (panels a and b) and in the presence (panels c and d) of human lactoferrin (LF). ( a ) Signals of the binding and dissociation experiment performed via BLI with the protein system RBD and ACE2. ACE2 was loaded on ProA biosensors, and RBD was in solution at decreasing concentrations. The vertical dashed lines indicate the time interval of the binding step (180 s) and of the dissociation step (120 s). ( b ) Aggregation signals from turbidimetric assays. Latex nanospheres were coated with the ACE2 protein and mixed in solution with RBD at decreasing concentrations. Panels ( c , d ): Time-courses of the reaction with RBD and ACE2 in the presence of LF in solution. ( c ) BLI signals of LF in solution with RBD as the analyte protein at different LF concentrations, with ACE2 loaded on ProA biosensors. The vertical dashed lines indicate the start of the binding step (180 s) and of the dissociation step (120 s). ( d ) Turbidimetry assay performed with LF at variable concentrations and RBD present at a fixed concentration, in solution with latex nanospheres coated with ACE2.

Article Snippet: Human milk lactoferrin (LF) protein was from Sigma-Aldrich, with molecular weight between 82.4 and 87 kD; the estimated extinction coefficient was ε mM = 110.96 mM −1 cm −1 (280 nm), and the pI was 8.7.

Techniques: Binding Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Lactoferrin Inhibition of the Complex Formation between ACE2 Receptor and SARS CoV-2 Recognition Binding Domain

doi: 10.3390/ijms23105436

Figure Lengend Snippet: Inhibition of RBD−ACE2 complex formation in the presence of increasing concentrations of lactoferrin. The turbidimetric data of d were used to analyze the effect of increasing lactoferrin concentration (in log scale). The absorbance amplitudes of the reactions are shown in panel ( a ), whereas the initial rates of the same curves are depicted in panel ( b ). Interpolating red curves represent the best fit to the data obtained by Equation (S8). The data analysis, performed via custom MATLAB program, yielded apparent K obs values of 9.5 ± 1.5 µM ( a ) and 6.3 ± 1.2 µM ( b ), respectively, with cooperativity coefficient n of 1.44 ± 0.11 and 1.54 ± 0.08, respectively.

Article Snippet: Human milk lactoferrin (LF) protein was from Sigma-Aldrich, with molecular weight between 82.4 and 87 kD; the estimated extinction coefficient was ε mM = 110.96 mM −1 cm −1 (280 nm), and the pI was 8.7.

Techniques: Inhibition, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Lactoferrin Inhibition of the Complex Formation between ACE2 Receptor and SARS CoV-2 Recognition Binding Domain

doi: 10.3390/ijms23105436

Figure Lengend Snippet: Regions of high binding propensity of ACE2 to LF. ( a ) Binding propensity of human ACE2 residues for human lactoferrin obtained on the basis of local shape complementarity of the molecular surfaces . Only residues whose binding propensity is higher than 0.85 are reported. Blue, orange, and green bands highlight three different portions of the molecular surfaces characterized by high binding propensity. ( b ) Molecular surface of the extracellular region of human ACE2 colored according to the Zernike binding propensity score. The color turns from white to red as the local binding propensity increases. The surface was oriented to show the region around residue Tpr48, which is comprised in the blue band in panel ( a ). ( c ) Same as in ( b ) but displaying the region around residue Arg177 and marked with the orange bands; a cartoon representation of the RDB of the spike protein bound to ACE2 is also shown. ( d ) Same as in ( b ), but for the green band of the panel ( a ) marking the region around residue Asn572.

Article Snippet: Human milk lactoferrin (LF) protein was from Sigma-Aldrich, with molecular weight between 82.4 and 87 kD; the estimated extinction coefficient was ε mM = 110.96 mM −1 cm −1 (280 nm), and the pI was 8.7.

Techniques: Binding Assay