Contamination of surface water and drinking water with pharmaceuticals presents an environmental concern. It has been shown to affect aquatic organisms and have adverse health effects on humans. One of the most common pharmaceutical contaminants is the opioid analgesic tramadol. In this communication, we report on the first surface plasmon resonance biosensor-based detection of tramadol in water. The biosensor utilizes a binding inhibition format and enables detection of tramadol at a wide range of concentrations (5 orders of magnitude) with a limit of detection of 0.52 µg/L. The results of a small-scale environmental study are reported in which the biosensor was used to analyze river water samples. The results were found to agree well with those obtained using the liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).

• Keywords
• Biosensor, Surface plasmon resonance, Tramadol, Water quality monitoring,
• MeSH
• Biosensing Techniques * methods   MeSH
• Water Pollutants, Chemical * analysis   MeSH
• Limit of Detection MeSH
• Environmental Monitoring * methods   MeSH
• Analgesics, Opioid * analysis   MeSH
• Surface Plasmon Resonance * methods   MeSH
• Rivers chemistry   MeSH
• Tandem Mass Spectrometry MeSH
• Tramadol * analysis   MeSH
• Chromatography, High Pressure Liquid MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Water Pollutants, Chemical * MeSH
• Analgesics, Opioid * MeSH
• Tramadol * MeSH

Aberrant glycosylation of glycoproteins has been linked with various pathologies. Therefore, understanding the relationship between aberrant glycosylation patterns and the onset and progression of the disease is an important research goal that may provide insights into cancer diagnosis and new therapy development. In this study, we use a surface plasmon resonance imaging biosensor and a lectin array to investigate aberrant glycosylation patterns associated with oncohematological disease-myelodysplastic syndromes (MDS). In particular, we detected the interaction between the lectins and glycoproteins present in the blood plasma of patients (three MDS subgroups with different risks of progression to acute myeloid leukemia (AML) and AML patients) and healthy controls. The interaction with lectins from Aleuria aurantia (AAL) and Erythrina cristagalli was more pronounced for plasma samples of the MDS and AML patients, and there was a significant difference between the sensor response to the interaction of AAL with blood plasma from low and medium-risk MDS patients and healthy controls. Our data also suggest that progression from MDS to AML is accompanied by sialylation of glycoproteins and increased levels of truncated O-glycans and that the number of lectins that allow discriminating different stages of disease increases as the disease progresses.

• MeSH
• Leukemia, Myeloid, Acute * MeSH
• Biosensing Techniques * MeSH
• Glycoproteins metabolism   MeSH
• Glycosylation MeSH
• Plasma metabolism   MeSH
• Lectins MeSH
• Humans MeSH
• Myelodysplastic Syndromes * therapy   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Glycoproteins MeSH
• Lectins MeSH

Base excision repair is one of the important DNA repair mechanisms in cells. The fundamental role in this complex process is played by DNA glycosylases. Here, we present a novel approach for the real-time measurement of uracil DNA glycosylase activity, which employs selected oligonucleotides immobilized on the surface of magnetic nanoparticles and Förster resonance energy transfer. We also show that the approach can be performed by surface plasmon resonance sensor technology. We demonstrate that the immobilization of oligonucleotides provides much more reliable data than the free oligonucleotides including molecular beacons. Moreover, our results show that the method provides the possibility to address the relationship between the efficiency of uracil DNA glycosylase activity and the arrangement of the used oligonucleotide probes. For instance, the introduction of the nick into oligonucleotide containing the target base (uracil) resulted in the substantial decrease of uracil DNA glycosylase activity of both the bacterial glycosylase and glycosylases naturally present in nuclear lysates.

• Keywords
• Förster resonance energy transfer, base excision repair, immobilized oligonucleotides, surface plasmon resonance, uracil DNA glycosylase,
• MeSH
• Cell Nucleus metabolism   MeSH
• Humans MeSH
• Magnetic Iron Oxide Nanoparticles MeSH
• Oligonucleotide Probes chemistry   metabolism   MeSH
• DNA Repair MeSH
• Fluorescence Resonance Energy Transfer MeSH
• Uracil-DNA Glycosidase metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Oligonucleotide Probes MeSH
• Uracil-DNA Glycosidase MeSH

Myelodysplastic syndromes (MDS) are a heterogeneous group of hematological malignancies with a high risk of transformation to acute myeloid leukemia (AML). MDS are associated with posttranslational modifications of proteins and variations in the protein expression levels. In this work, we present a novel interactomic diagnostic method based on both protein array and surface plasmon resonance biosensor technology, which enables monitoring of protein-protein interactions in a label-free manner. In contrast to conventional methods based on the detection of individual biomarkers, our presented method relies on measuring interactions between arrays of selected proteins and patient plasma. We apply this method to plasma samples obtained from MDS and AML patients, as well as healthy donors, and demonstrate that even a small protein array comprising six selected proteins allows the method to discriminate among different MDS subtypes and healthy donors.

• MeSH
• Principal Component Analysis MeSH
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Protein Interaction Mapping * MeSH
• Young Adult MeSH
• Myelodysplastic Syndromes blood   diagnosis   MeSH
• Surface Plasmon Resonance MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Protein Binding MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The scavenger receptor cysteine-rich (SRCR) family comprises a group of membrane-attached or secreted proteins that contain one or more modules/domains structurally similar to the membrane distal domain of type I macrophage scavenger receptor. Although no all-inclusive biological function has been ascribed to the SRCR family, some of these receptors have been shown to recognize pathogen-associated molecular patterns (PAMP) of bacteria, fungi, or other microbes. SSc5D is a recently described soluble SRCR receptor produced by monocytes/macrophages and T lymphocytes, consisting of an N-terminal portion, which contains five SRCR modules, and a large C-terminal mucin-like domain. Toward establishing a global common role for SRCR domains, we interrogated whether the set of five SRCR domains of SSc5D displayed pattern recognition receptor (PRR) properties. For that purpose, we have expressed in a mammalian expression system the N-terminal SRCR-containing moiety of SSc5D (N-SSc5D), thus excluding the mucin-like domain likely by nature to bind microorganisms, and tested the capacity of the SRCR functional groups to physically interact with bacteria. Using conventional protein-bacteria binding assays, we showed that N-SSc5D had a superior capacity to bind to Escherichia coli strains RS218 and IHE3034 compared with that of the extracellular domains of the SRCR proteins CD5 and CD6 (sCD5 and sCD6, respectively), and similar E. coli-binding properties as Spα, a proven PRR of the SRCR family. We have further designed a more sensitive, real-time, and label-free surface plasmon resonance (SPR)-based assay and examined the capacity of N-SSc5D, Spα, sCD5, and sCD6 to bind to different bacteria. We demonstrated that N-SSc5D compares with Spα in the capacity to bind to E. coli and Listeria monocytogenes, and further that it can distinguish between pathogenic E. coli RS218 and IHE3034 strains and the non-pathogenic laboratory E. coli strain BL21(DE3). Our work thus advocates the utility of SPR-based assays as sensitive tools for the rapid screening of interactions between immune-related receptors and PAMP-bearing microbes. The analysis of our results suggests that SRCR domains of different members of the family have a differential capacity to interact with bacteria, and further that the same receptor can discriminate between different bacteria strains and species.

• Keywords
• bacteria, pattern recognition receptors, scavenger receptor cysteine-rich, surface plasmon resonance,
• Publication type
• Journal Article MeSH