Portable and Quantitative Detection of Protein Biomarkers and Small Molecular Toxins Using Antibodies and Ubiquitous Personal Glucose Meters

Developing portable and low-cost methods for quantitative detection of large protein biomarkers and small molecular toxins can play a significant role in controlling and preventing diseases or toxins outbreaks. Despite years of research, most current methods still require laboratory-based or customized devices that are not widely available to the general public for quantitative analysis. We have previously demonstrated the use of personal glucose meters (PGMs) and functional DNAs for the detection of many nonglucose targets. However, the range of targets detectable by functional DNAs is limited at the current stage. To expand the range of targets that can be detected by PGMs, we report here the use of antibodies in combination with sandwich and competitive assays for quantitative detection of protein biomarkers (PSA, with a detection limit of 0.4 ng/mL) and small molecular toxins (Ochratoxin A, with a detection limit of 6.8 ng/mL), respectively. In both assay methods, with invertase conjugates as the link, quantitative detection is achieved via the dependence between the concentrations of the targets in the sample and the glucose measured by PGMs. Given the wide availability of antibodies for numerous targets, the methods demonstrated here can expand the range of target detection by PGMs significantly

MicroSatResults

Symbiodinium genotypes associated with Aiptasia. Genotypes were determined using 18S RFLP, ITS2 DGGE, and 6 microsatellite markers specific to Symbiodinium Clade B. The spreadsheets provide sample location, sample ID, RFLP and DGGE proflies, as well as the allele sizes for each microsatellite. The data are presented on two worksheets within the file. The first worksheet includes samples in which Clade B was detected according to the 18S RFLP. The second worksheet includes samples from Florida in which only Clade A was detected according to the 18S RFLP, but the samples were screened with the Clade B-specific microsatellites to determine whether low levels of Clade B (i.e., "background" or "cryptic" populations) could be detected

TfOH- and HBF₄‑Mediated Formal Cycloisomerizations and [4+3] Cycloadditions of Allene-alkynylbenzenes

A metal-free, TfOH (1.1 equiv)-mediated formal cycloisomerization of easily prepared allene-alkynylbenzenes to give pyrrolidines and cyclopentanes derivatives was developed. This reaction is initiated by the generation of allylic cation from allene, followed by alkyne’s reaction with the allylic cation, to give a vinyl cation, which is finally intercepted by the triflate (TfO) anion. This cycloisomerization can be further tuned to become an acid-mediated intramolecular formal [4+3] cycloaddition by using 10 equiv of TfOH (The excess acid was used to promote the Friedel–Crafts reaction of the acid-mediated cycloisomerization products). The present system can also be applied to synthesized F-incorporated products by using HBF₄ or Me₃OBF₄ as the fluoro source

TfOH- and HBF₄‑Mediated Formal Cycloisomerizations and [4+3] Cycloadditions of Allene-alkynylbenzenes

A metal-free, TfOH (1.1 equiv)-mediated formal cycloisomerization of easily prepared allene-alkynylbenzenes to give pyrrolidines and cyclopentanes derivatives was developed. This reaction is initiated by the generation of allylic cation from allene, followed by alkyne’s reaction with the allylic cation, to give a vinyl cation, which is finally intercepted by the triflate (TfO) anion. This cycloisomerization can be further tuned to become an acid-mediated intramolecular formal [4+3] cycloaddition by using 10 equiv of TfOH (The excess acid was used to promote the Friedel–Crafts reaction of the acid-mediated cycloisomerization products). The present system can also be applied to synthesized F-incorporated products by using HBF₄ or Me₃OBF₄ as the fluoro source

Aiptasia_Sym_B_MicSat_Fla2

Nucleotide alignment of DNA sequences from microsatellite flanking region loci CA4.86 and Si15. The alignment includes Symbiodinium minutum associated with Aiptasia from field collected and cultured specimens as well as Symbiodinium samples collected from other hosts

CignobilisCmelampygusTOTALATPase6ATPase8COMBINED.nex

Mitochondrial ATPase 6 and ATPase 8 DNA sequences utilized in the study in NEXUS format. Also see README file

Simple and Efficient Method to Purify DNA–Protein Conjugates and Its Sensing Applications

DNA–protein conjugates are very useful in analytical chemistry for target recognition and signal amplification. While a number of methods for conjugating DNA with proteins are known, methods for purification of DNA–protein conjugates from reaction mixture containing unreacted proteins are much less investigated. In this work, a simple and efficient approach to purify DNA–invertase conjugates from reaction mixture via a biotin displacement strategy to release desthiobiotinylated DNA–invertase conjugates from streptavidin-coated magnetic beads was developed. The conjugates purified by this approach were utilized for quantitative detection of cocaine and DNA using a personal glucose meter through structure-switching DNA aptamer sensors and competitive DNA hybridization assays, respectively. In both cases, the purified DNA–invertase conjugates showed better performance compared to the same assays using unpurified conjugates. The approach demonstrated here can be further expanded to other DNA and proteins to generate purified DNA–protein conjugates for analytical and other applications

Forest plot of the subgroup analysis of Tai Chi for fatigue based on different duration.

<p>Forest plot of the subgroup analysis of Tai Chi for fatigue based on different duration.</p

Meta-analysis of Tai Chi for fatigue.

<p>A random effect model was performed to test for high statistical heterogeneity. Subgroup analysis was based on three different conditions including cancer, multiple sclerosis and age-related fatigue. Only descriptive analysis was performed for Tai Chi for rheumatoid arthritis, primary insomnia and COPD related fatigue.</p

Characteristics of included studies.

<p>Characteristics of included studies.</p