A rapid solid-phase fluorimetric assay for measuring bacterial adherence, using DNA-binding stains

Bosch, Jos A.; Veerman, E.C.I.; Turkenburg, Marjolein; Hartog, K.; Bolscher, Jan G. M.; Amerongen, A. van Nieuw

doi:10.1016/s0167-7012(02)00220-8

Cited by 13 publications

(7 citation statements)

References 16 publications

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“…Before assaying saliva was centrifuged at 2000g for 5 min. Salivary alpha-amylase measurement was completed using an enzyme kinetic method as described previously (Bosch et al, 2003; Rohlder & Nater, 2009). Saliva was diluted at 1:625 with ultrapure water, and diluted saliva was incubated with substrate reagent (α-amylase EPS Sys; Indianapolis, IN) at 37C for 3 min before a first absorbance reading was taken at 405 nm with a Tecan Sunrise ELISA reader (Tecan, Morrisville, NC).…”

Section: Methodsmentioning

confidence: 99%

Self-compassionate young adults show lower salivary alpha-amylase responses to repeated psychosocial stress

et al. 2015

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In this study we tested the hypothesis that participants higher in dispositional self-compassion would show lower stress-induced reactivity of salivary alpha-amylase (sAA), a marker of sympathetic nervous system activation. Thirty-three healthy participants (18–34 years old) were exposed to a standardized laboratory stressor on two consecutive days. Self-compassion, self-esteem, and demographic factors were assessed by questionnaire and sAA was assessed at baseline and at 1, 10, 30, and 60 minutes following each stressor. Self-compassion was a significant negative predictor of sAA responses on both days. This relationship remained significant when controlling for self-esteem, subjective distress, age, gender, ethnicity, and Body Mass Index (BMI). These results suggest that self-compassion may serve as a protective factor against stress-induced physiological changes that have implications for health.

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Section: Methodsmentioning

confidence: 99%

Self-compassionate young adults show lower salivary alpha-amylase responses to repeated psychosocial stress

et al. 2015

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“…Adhesion assays were performed as described previously (Leito et al, 2008) according to the method of Bosch et al (2003). After an o/n culture of C. albicans in Sabouraud dextrose broth, yeast cells were harvested by centrifugation (3000 g at 22 1C for 5 min), washed two times in TBST (10 mM Tris, 150 mM NaCl, 0.1% Tween 20, pH 7.4) and suspended in TBST-Ca (TBST with 2.5 mM CaCl 2 ) to a density of 1 Â 10 7 cells mL À1 .…”

Section: Adhesion Assaymentioning

confidence: 99%

Identification of salivary components that induce transition of hyphae to yeast inCandida albicans

Leito

Ligtenberg

Nazmi

et al. 2009

FEMS Yeast Research

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Candida albicans, the major human fungal pathogen, undergoes a reversible morphological transition from single yeast cells to pseudohyphae and hyphae filaments. The hyphae form is considered the most invasive form of the fungus. The purpose of this study is to investigate the effect of saliva on hyphae growth of C. albicans. Candida albicans hyphae were inoculated in Roswell Park Memorial Institute medium with whole saliva, parotid saliva or buffer mimicking the saliva ion composition, and cultured for 18 h at 37 degrees C under aerobic conditions with 5% CO(2). Whole saliva and parotid saliva induced transition to yeast growth, whereas the culture with buffer remained in the hyphae form. Parotid saliva was fractionated on a reverse-phase C8 column and each fraction was tested for inducing transition to yeast growth. By immunoblotting, the salivary component in the active fraction was identified as statherin, a phosphoprotein of 43 amino acids that has been implicated in remineralization of the teeth. Synthetically made statherin induced transition of hyphae to yeast. By deletion of five amino acids at the negatively charged N-terminal site (DpSpSEE), yeast-inducing activity and binding to C. albicans were increased. In conclusion, statherin induces transition to yeast of C. albicans hyphae and may thus contribute to the oral defense against candidiasis.

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“…Adhesion Assays-Bacterial adhesion was examined using a microtiter plate method based on the labeling of microorganisms with cellpermeable DNA binding probes, as reported earlier (17,35). Fluotrac 600 microtiter plates (Greiner, Recklinghausen, Germany) were coated with various amounts of either synthetic peptides or purified DMBT1 SAG .…”

Section: Fig 1 Determination Of the Minimal Binding Sequence Withinmentioning

confidence: 99%

Bacteria Binding by DMBT1/SAG/gp-340 Is Confined to the VEVLXXXXW Motif in Its Scavenger Receptor Cysteine-rich Domains

Bikker

Ligtenberg

End

et al. 2004

Journal of Biological Chemistry

116

130

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The scavenger receptor cysteine-rich (SRCR) proteins form an archaic group of metazoan proteins characterized by the presence of SRCR domains. These proteins are classified in group A and B based on the number of conserved cysteine residues in their SRCR domains, i.e. six for group A and eight for group B. The protein DMBT1 (deleted in malignant brain tumors 1), which is identical to salivary agglutinin and lung gp-340, belongs to the group B SRCR proteins and is considered to be involved in tumor suppression and host defense by pathogen binding. In a previous study we used nonoverlapping synthetic peptides covering the SRCR consensus sequence to identify a 16-amino acid bacteriabinding protein loop (peptide SRCRP2; QGRVEVLYRG-SWGTVC) within the SRCR domains. In this study, using overlapping peptides, we pinpointed the minimal bacteria-binding site on SRCRP2, and thus DMBT1, to an 11-amino acid motif (DMBT1 pathogen-binding site 1 or DMBT1pbs1; GRVEVLYRGSW). An alanine substitution scan revealed that VEVL and Trp are critical residues in this motif. Bacteria binding by DMBT1pbs1 was different from the bacteria binding by the macrophage receptor MARCO in which an RXR motif was critical. In addition, the homologous consensus sequences of a number of SRCR proteins were synthesized and tested for bacteria binding. Only consensus sequences of DMBT1 orthologues bound bacteria by this motif.The scavenger receptor cysteine-rich (SRCR) 1 proteins form an archaic group of metazoan proteins (1-5). This group of glycoproteins comprises cell surface molecules as well as secreted proteins that are characterized by the presence of one or more SRCR domains. SRCR domains consist of ϳ110 amino acids and are divided into groups A and B based on the number of conserved cysteine residues, namely six for group A and eight for group B.The best studied members of the group A SRCR proteins are the macrophage scavenger receptor (MSR1), the Mac 2-binding protein (Mac-2bp), and MARCO. Both MSR1 and MARCO are known to interact with bacteria (6, 7). In contrast to MARCO (8), the SRCR domain of MSR1 does not seem to be involved in bacteria binding (9, 10). Bacteria binding by MARCO involves an RXR motif within the SRCR domain, indicating that ionic interactions play a crucial role in the interaction with its negatively charged ligands (6).Group B SRCR proteins are generally involved in the regulation of cellular immune responses. In vertebrates, the group B SRCR proteins can be divided, on the basis of their structure and sequence homology, into three subgroups (11). The first subgroup includes CD5 (12), CD6 (13), and SP␣ (14). CD5 and CD6 are composed of an extracellular region of three SRCR domains, a transmembrane region, and a cytoplasmic region. SP␣ lacks the latter two regions but contains three SRCR domains that are highly homologous to those of CD5 and CD6. These three proteins are mainly expressed by T-cells and Bcells (12, 13). The second subgroup of SRCR group B molecules is the workshop cluster 1 (WC1) family, which includes WC1, ...

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A rapid solid-phase fluorimetric assay for measuring bacterial adherence, using DNA-binding stains

Cited by 13 publications

References 16 publications

Self-compassionate young adults show lower salivary alpha-amylase responses to repeated psychosocial stress

Self-compassionate young adults show lower salivary alpha-amylase responses to repeated psychosocial stress

Identification of salivary components that induce transition of hyphae to yeast inCandida albicans

Bacteria Binding by DMBT1/SAG/gp-340 Is Confined to the VEVLXXXXW Motif in Its Scavenger Receptor Cysteine-rich Domains

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