Correlation of cortisol in 1-cm hair segment with salivary cortisol in human: hair cortisol as an endogenous biomarker.

BACKGROUND: Cortisol level in human hair would be an endogenous biomarker for the retrospective assessment of long-term central hypothalamo-pituitary-adrenal activity. However, no direct evidence supports that blood-related diffusion is a biologically endogenous source of hair cortisol in humans. The present study aims to validate the direct correlation between cortisol in 1-cm hair segments and salivary cortisol in healthy humans. METHODS: We collected three saliva samples from the same participant at Time 1, Time 2 (1 week later) and Time 3 (2 weeks later), and hair 4 weeks later. Cortisol levels in 1-cm hair segments and saliva were determined with high performance liquid chromatography tandem mass spectrometry. RESULTS: Salivary cortisol at Time 1 was significantly associated with that at Time 2 (r=0.514, p=0.003), but not with that at Time 3 (r=0.187, p=0.305); and the one at Time 2 was significantly associated with that at Time 3 (r=0.380, p=0.032). Hair cortisol was significantly correlated with salivary cortisol at Time 2 (r=0.389, p<0.05) and average salivary cortisol (r=0.383, p<0.05) from three sampling. CONCLUSIONS: Our results confirmed that blood-related diffusion mechanism is a biologically endogenous source of hair cortisol.

High-Strength Hydrogel Adhesive Formed via Multiple Interactions for Persistent Adhesion under Saline.

Hydrogel adhesives have been widely used in wet environments. Nonetheless, strong and stable persistent adhesion remains a challenge. Here, we report a facile yet powerful strategy to construct high-strength hydrogel adhesives for durable adhesion in a saline environment. Such a hydrogel consists of two polymer networks: a hydrophobic-associated polyacrylamide network of covalent and noncovalent cross-links and an alginate network cross-linked by divalent cations in saline. Meanwhile, polydopamine nanoparticles formed through in-situ self-polymerization are distributed evenly throughout the system to provide underwater adhesion. A low and controllable swelling rate and high compressive strength of hydrogels can be achieved via this multiple interaction strategy. Ultimately, this strategy contributes to the persistent underwater adhesion of hydrogels, and the decreasing rate of lap-shear adhesion strength of hydrogels is only 24.79 ± 8.01% after saline immersion for up to 21 days. Moreover, good cytocompatibility of hydrogels is helpful for their application in the biomedical field.

Application of nanofiber-packed SPE for determination of salivary-free cortisol using fluorescence precolumn derivatization and HPLC detection.

Salivary cortisol has emerged as an easy-to-collect biologic marker of stress in many researches. In this study, we present a method for the determination of salivary-free cortisol using HPLC method with fluorescence precolumn derivatization, which is based on a novel extraction from the strongly acidic medium (fluorescent derivatives of cortisol in sulfuric acid medium) by electrospun polystyrene nanofibers packed SPE. For high-throughput sample extraction, an array pretreatment device based on nanofibers packed SPE micro-column was designed. The LOD of cortisol was 0.01 microg/L (S/N=3). The RSDs (n=6) for all analytes were below 8.0%, and the recoveries were 110, 102.4, and 99.4% (n=3) for saliva spiked with 0.1, 10, and 20 microg/L of cortisol, respectively. The proposed method was then successfully applied in the determination of free cortisol in human saliva. The salivary cortisol concentrations in the real samples ranged from 0.22 to 7.45 microg/L. The nanofiber-packed SPE overcame the low extraction recovery and bad clean-up effect of the conventional methods, and increased the sensitivity and selectivity of the method.

Mussel-inspired hybrid network hydrogel for continuous adhesion in water.

The mussel-inspired catechol-based strategy has been widely used in the development of adhesives. However, the properties of the obtained adhesives were still severely limited in a humid environment, particularly in water. In this study, a facile and versatile approach was proposed to prepare an underwater adhesion hydrogel. First, dopamine (DA) was grafted on oxidized carboxymethylcellulose (OCMC) to obtain dopamine-grafted oxidized carboxymethylcellulose (OCMC-DA). Second, the acrylamide (AM) monomer was conjugated with OCMC-DA by a Schiff base reaction, and then polymerized to form an OCMC-DA/PAM hydrogel. The properties of the resulting hydrogel have been fully characterized. The underwater adhesion strength of the hydrogel can reach as high as 86.3 ± 7.2 kPa and reduced to 43 ± 3.4 kPa after being immersed in water for 9 days. More remarkably, we found that the maximal adhesion strength was shown when the G' and G'' of the hydrogel were very close. Moreover, we demonstrated the mechanical properties of our fabricated hydrogel by compressive tests and rheological analysis. The adhesive hydrogel also exhibits excellent biocompatibility.

A high-strength double network polydopamine nanocomposite hydrogel for adhesion under seawater.

Mussel-inspired catechol-based strategy has been widely used in the development of underwater adhesives. Nonetheless, the properties of the adhesives were still severely limited under harsh environments. A facile approach was proposed herein to prepare a double network hydrogel adhesive with low swelling rate and high strength in seawater, where the first network was polyacrylamide (PAM) and the second network was alginate (Alg). Meanwhile, polydopamine (PDA) nanoparticles, which were formed through self-polymerization as adhesion anchoring sites, distributed evenly throughout the double network hydrogel and effectively enhanced the adhesion capability of the hydrogel. The properties of the resulting hydrogel have been fully characterized. The optimal adhesion strength of the hydrogel adhesive in seawater was as high as 146.84 ± 7.78 kPa. Furthermore, the hydrogel also has excellent ability to promote the growth of zooxanthellae. Our studies provide useful insights into the rational design of underwater adhesives with high performances even beyond nature.

On the origin of vertebrate body plan: Insights from the endoderm using the hourglass model.

The vertebrate body plan is thought to be derived during the early Cambrian from a worm-like chordate ancestor. While all three germ layers were clearly involved in this innovation, the role of the endoderm remains elusive. According to the hourglass model, the optimal window for investigating the evolution of vertebrate endoderm-derived structures during cephalochordate development is from the Spemann's organizer stage to the opening of the mouth (Stages 1-7, described herein). Regulatory gene expression, examined during these stages, illustrate that the cephalochordate endoderm is patterned into 12 organ primordia. Early vertebrates inherited at least a portion of 6 of these primordia, while the remainder were lost. Of those that were preserved, we demonstrate that the vertebrate symmetric mouth was built on a vestige of the anterior pre-oral pit, that the pre-existing pharyngeal pouch in this chordate ancestor laid the foundation for the new neural crest cell (NCC)-derived vertebrate-type pharyngeal arches, that the thyroid evolved from the posterior endostyle primordim, that the pancreas was derived from the Pdx1-expressing diverticulum primordium, and the small and large intestines originated with the Cdx1-expressing hindgut rudiments. This investigation uncovers the evolutionary foundations of vertebrate endoderm-derived structures, and demonstrates that the number of organ primordia were reduced during evolution.

PCR-product microarray based on polyacrylic acid-modified surface for SNP genotyping.

PCR-product microarray has great efficiency in SNP genotyping, mutation screening and epigenetic analyzing from a large number of samples. The current PCR-product microarray technology is mostly based on the 3-D gel microarray technologies due to its high loading capacity for PCR products, while there is little progress for PCR-product microarray on planar glass, which gives low background and convenient fabrication. In this study, we improved the PCR-product microarray on planar glass by employing a polyacrylic acid-covered slide. The raw amino-modified PCR products were simply precipitated with ethanol and directly spotted for DNA immobilization. Three detection methods of hybridization, solid-single base extension and solid-multiple bases elongation were carried out for single nucleotide variation identification on the PCR-product microarray. The experimental results showed that the high immobilization yield for raw PCR product was achieved, and the high specificity and high ratio of S/N for genotyping on the microarray were obtained. SNP genotyping of cytochrome P450 2D6 gene in 30 individuals was successfully demonstrated. This study has significantly increased the performances of PCR-product microarray, which could improve its applications in SNP genotyping and mutation screening in large number of individuals.

Ultrasonic approach to enhance signal-to-noise in three dimensional microarray.

Three-dimensional polyacrylamide gel microarray (3-D gel microarray) has a strong adsorption for hybridzation probes with fluorescent label, resulting in high fluorescent background noise. The background noise has hampered the broad application of the microarray in scientific research. A low-frequency ultrasound of 40 kHz has been successfully employed to efficiently remove the nonspecifically bound fluorescent probes from the 3-D gel microarray. The method can significantly enhance the signal to noise ratio of the 3-D gel microarray and make it work better in single-nucleotide polymorphism genotyping.

Polyacrylamide gel film immobilized molecular beacon array for single nucleotide mismatch detection.

We reported polyacrylamide gel immobilized molecular beacon array for single nucleotide mismatch detection in this paper. Molecular beacons are oligonucleotide probes fluorescing upon hybridization to their complementary DNA/RNA targets with excellent sensitivity and high selectivity. The specially designed molecular beacon for immobilization contains a 15 base loop sequence with a 5 base pair stem, a polyT (20 bases) spacer, a 5'-end amino group for immobilization, a fluorescein in the middle of the sequence as the fluorophore, and a 3'-end DABCYL as the quencher. Between the 5'-end amino group and the stem, the polyT is used to minimize disability caused by 5'-end immobilization. The molecular beacon microarray was fabricated by a pin-based spotting robot and the hybridization was investigated by confocal microscope. A real-time hybridization process at room temperature was registered every minute for 20 min after the target solution was pumped into the hybridization cell. The result indicates that a polyacrylamide film coated glass slide provides an ideal solution-like environment for molecular beacon probes. The potential applications of this kind of molecular beacon array are mutation detection, disease mechanisms, disease diagnostics, etc. in a parallel, cost saving, and label-free detection way.

Amperometric hydrogen peroxide biosensor with sol-gel/chitosan network-like film as immobilization matrix.

A new type of sol-gel/organic hybrid composite material based on the cross-linking of natural polymer chitosan with (3-aoryloxypropyl) dimethoxymethylsilane was developed for the fabrication of an amperometric H(2)O(2) biosensor. The composite film was used to immobilize horseradish peroxidase (HRP) on a gold disk electrode. The properties of sol-gel/chitosan and sol-gel/chitosan-HRP films have been carefully characterized by atomic force microscopy and Fourier transform infrared. By using fluorescent label, a protein density on sol-gel/chitosan has been calculated to be 3.14 x 10(12) moleculescm(-2). With the aid of catechol mediator, the biosensor had a fast response of less than 2 s with linear range of 5.0 x 10(-9)-1.0 x 10(-7) mol l(-1) and a detection limit of 2 x 10(-9) mol l(-1). Its current response shows a typical Michaelis-Menten mechanism. The apparent Michaelis-Menten constant K(M)(app) is found to be 1.30 micromol l(-1). The activation energy for enzymatic reaction is calculated to be 8.22 kJ mol(-1). The biosensor retained approximately 75% of its original activity after about 60 days of storage in a phosphate buffer at 4 degrees C.

Effect of polyethylene glycols on the alkaline-induced molten globule intermediate of bovine serum albumin.

In the present study, the formation of one molten globule-like unfolding intermediate of bovine serum albumin (BSA) at pH 11.2 has been established with the help of circular dichroism (CD) spectra, fluorescence spectroscopy and 'phase diagram' approach. Additionally, we have shown the conformational changes occurring in the pH 11.2 intermediate of BSA when it was exposed to different molecular weight polyethylene glycols (PEGs) at varying concentrations. When the pH 11.2 intermediate of BSA was treated by PEG 400 there was induction of secondary and non-native tertiary contacts. In case of PEG 8000 and PEG 20,000, the loss in secondary as well as tertiary structure was observed. PEG 8000 and 20,000 altered the conformation of the pH 11.2 intermediate and resulted in its transition to another intermediate state in which the hydrophobic patches were inaccessible.

Automated time-resolved analysis of bacteria-substrate interactions using functionalized microparticles and flow cytometry.

Surface biofouling poses an increasing problem in industrial and health care applications, driving research for surface coatings to prevent anti-microbial colonization and characterization of the efficacy of the same. The diversity and increasing sophistication of such coatings, which postulate different types of anti-microbial action on planktonic and surface adhering bacteria, challenge the suitability of current approaches to evaluate and compare the different approaches as well as the speed and accuracy at which screening can be made. We describe and provide proof of principle for a method to use microparticles functionalized with molecular coatings through self-assembly together with flow cytometry readout to evaluate Escherichia coli bacteria surface adhesion and killing efficiency. Advantages of the method are the automation of the method that allows recording an immense number of interactions and the possibility to simultaneously record effects on both surface adhering and planktonic bacteria. We demonstrate and discuss design criteria to obtain this information on two coatings, poly(L-lysine)-graft-C(3)H(6)N(+)(CH(3))(2)C(12)H(25) (PLL-g-QAC) and poly(L-lysine)-graft-poly(ethylene glycol)-C(3)H(6)N(+)(CH(3))(2)C(12)H(25) (PLL-g-PEG-QAC), which exemplify two different approaches to creating anti-microbial interfaces. Despite an apparent higher killing efficiency of the PLL-g-QAC during brief exposures, the rapid fouling of that surface quickly reduces its efficiency, whereas the PLL-g-PEG-QAC coating showed greater promise in reducing the growth and interfacial colonization of bacteria over longer time scales.

Influences of cationic, anionic, and nonionic surfactants on alkaline-induced intermediate of bovine serum albumin.

The formation of one molten globule-like intermediate of bovine serum albumin (BSA) at pH 11.2 has been established by circular dichroism (CD) spectra, fluorescence spectroscopy and 'phase diagram' approach. Three types of surfactants induced alpha-helical structure and resumed the tertiary structure of alkaline-induced BSA intermediate. The internalized tryptophans of the intermediate were exposed to a more polar environment in the presence of surfactants. Induction of both secondary and tertiary structures in alkaline-unfolded BSA was greater in the presence of 3.5 mM cetyltrimethylammonium bromide (CTAB) followed by 8.0 mM sodium dodecyl sulphate (SDS) and the least in 0.1 mM Tween-20. Maximum ANS binding was obtained in the presence of 0.1 mM Tween-20 followed by 8.0 mM SDS. The addition of 3.5 mM CTAB resulted in loss of ANS binding sites, suggesting the burial of hydrophobic patches.

Genome-wide pathways analysis of nickel ion-induced differential genes expression in fibroblasts.

To reveal molecular mechanisms of the interaction between Ni2+ and cells, cDNA microarray technology and GenMAPP analysis were utilized to investigate changes of gene expression profile and identify significant biological pathways in mouse fibroblast cells (L-929) treated by 100 microm Ni2+ for 12, 24, 48 and 72 h, respectively. The microarray data was validated by real-time PCR. Methylthiazoltetrazolium (MTT) analysis and flow cytometry experiment were used to assess the cellular response of L-929 cells to Ni2+. It was found that six main biological pathways were affected by Ni2+ with 118 differentially expressed genes involved. Further analysis illuminated that the exposure of cells to Ni2+ may evoke series of cellular responses to hypoxia by regulating hypoxia-inducible gene expression and cause irreversible DNA damage. Cell cycle pathway analysis results showed DNA replication in S phase could be inhibited by Ni2+ which was consistent with the data gained from flow cytometry experiment. Compared to previous researches based on conventional molecular biology experiments, the present work has not only indirectly validated the findings of other groups but also obtained several discoveries related to cell-Ni2+ interaction, such as inhibition of electron transport chain and accumulation of extracellular matrix (ECM) collagens. The routine of the present study not only can analyze gene expression profile but also may provide a more convenient and efficient approach to explain molecular mechanisms of cell-biomaterial interaction.