Enhanced electrical conductivity of anticorrosive coatings by functionalized carbon nanotubes: effect of hydrogen bonding.

Carbon nanotubes and nanofibers (CNFs) are well-known nano additives to produce coating materials with high electrical and thermal conductivity and corrosion resistance. In this paper, coating materials incorporating hydrogen bonding offered significantly lower electrical resistance. The hydrogen bonding formed between functionalized carbon nanotubes and ethanol helped create a well-dispersed carbon nanotube network as the electron pathways. Electrical resistivity as low as 6.8 Ω cm has been achieved by adding 4.5 wt% functionalized multiwalled carbon nanotubes (MWNT-OH) to 75%polyurethane/25%ethanol. Moreover, the thermal conductivity of polyurethane was improved by 332% with 10 wt% addition of CNF. Electrochemical methods were used to evaluate the anti-corrosion properties of the fabricated coating materials. 75%polyurethane/25%ethanol with the addition of 3.0 wt% of MWNT-OH showed an excellent corrosion rate of 5.105 × 10-3mm year-1, with a protection efficiency of 99.5% against corrosive environments. The adhesion properties of the coating materials were measured following ASTM standard test methods. 75%polyurethane/25%ethanol with 3.0 wt% of MWNT-OH belonged to class 5 (ASTM D3359), indicating the outstanding adhesion of the coating to the substrate. These nanocoatings with enhanced electrical, thermal, and anti-corrosion properties consist of a choice of traditional coating materials, such as polyurethane, yielding coating durability with the ability to tailor the electrical and thermal properties to fit the desired application.

Diet, Physical Activity, and Screen Time to Sleep Better: Multiple Mediation Analysis of Lifestyle Factors in School-Aged Children with and without Attention Deficit Hyperactivity Disorder.

OBJECTIVE: This study examined the mediation roles of multiple lifestyle factors in school-aged children. Structural equation modeling (SEM) tested how lifestyle factors play mechanism roles one another in the impact of ADHD to seek theoretical and intervention insights. METHOD: An online survey assessed children's lifestyle factors including diet, physical activity, screen time, sleep difficulties, and having ADHD diagnosis. A multi-country sample from English speaking nations included 309 caregivers. Multiple regression and SEM were planned to identify significant correlates and mediators of ADHD in explaining lifestyle differences. RESULTS: Preliminary multiple regression showed only sleep quality was significantly different between children with and without ADHD. Significant triple mediation effects suggested diet, physical activity, and screen time mediated the ADHD impact on sleep quality. CONCLUSION: Researchers and practitioners may incorporate the findings to develop intervention models for children with ADHD attending to the mediational roles of lifestyle factors to improve sleep quality.

A New Automated Technology for Cerebrospinal Fluid Cell Counts: Comparison of Accuracy and Clinical Impact of GloCyte, Sysmex XN, and Manual Methods.

OBJECTIVES: The purpose of the study was to compare the performance of GloCyte (Advanced Instruments, Norwood, MA), a new semiautomated instrument for cerebrospinal fluid cell counting, with the manual hemocytometer method and the automated Sysmex XN (Sysmex, Kobe, Japan) body fluid mode. The clinical impact of replacing the manual method with either automated method was determined. METHODS: Fifty-seven samples from 38 patients were analyzed by all three methods. Pearson correlation and Passing-Bablok regression were used to compare methods. Cytospin smears were reviewed on all samples, and clinical histories were obtained. RESULTS: There was a strong linear relationship between the manual and automated methods for WBC counts ( R = 0.988 for GloCyte; R = 0.980 for Sysmex XN). Positive bias was absent or negligible for WBC counts less than 30/µL. GloCyte and manual RBC counts were equivalent. There were no samples for which replacement of manual WBC counts by automated counts would have changed the diagnosis. Both automated methods showed improved precision for WBC counts compared with the manual method. CONCLUSIONS: Replacing manual WBC counts by GloCyte or Sysmex XN WBC counts would improve consistency of results without compromising diagnostic accuracy.

Interaction between carbon nanotubes and mammalian cells: characterization by flow cytometry and application.

We show herein that CNT-cell complexes are formed in the presence of a magnetic field. The complexes were analyzed by flow cytometry as a quantitative method for monitoring the physical interactions between CNTs and cells. We observed an increase in side scattering signals, where the amplitude was proportional to the amount of CNTs that are associated with cells. Even after the formation of CNT-cell complexes, cell viability was not significantly decreased. The association between CNTs and cells was strong enough to be used for manipulating the complexes and thereby conducting cell separation with magnetic force. In addition, the CNT-cell complexes were also utilized to facilitate electroporation. We observed a time constant from CNT-cell complexes but not from cells alone, indicating a high level of pore formation in cell membranes. Experimentally, we achieved the expression of enhanced green fluorescence protein by using a low electroporation voltage after the formation of CNT-cell complexes. These results suggest that higher transfection efficiency, lower electroporation voltage, and miniaturized setup dimension of electroporation may be accomplished through the CNT strategy outlined herein.

Molecular monitoring of the intestinal flora by denaturing high performance liquid chromatography.

Gut flora analysis is hampered by the complexity of the intestinal microbiota and by inherent limitations of culture-based approaches. Therefore, culture-independent molecular methods based upon 16S rRNA gene analysis were applied successfully for the analysis of complex microbial communities. However, generally accepted and validated profiling methods such as denaturing and temperature gradient gel electrophoresis (DGGE/TGGE) are still laborious and time consuming. Thus, we adapted the separation of amplified bacterial 16S rRNA gene fragments by denaturing high performance liquid chromatography (DHPLC) using the WAVE Microbial Analysis System as a rapid and convenient means to display complex intestinal bacterial communities and to monitor changes in the gut flora. The separation of 16S rRNA gene fragments amplified from reference strains representing main gut bacterial populations and from human stool samples revealed that DHPLC analysis effectively detects bacterial groups predominant in the human gut flora. The investigation of faecal samples from hospitalized patients before, during and after antibiotic therapy showed that PCR-based DHPLC can be used to monitor gut flora changes. Results from DHPLC analysis were comparable with DGGE profiles generated from the same samples, demonstrating that the adapted DHPLC protocol is well suited for the analysis of complex microbial communities.

Use of denaturing high-performance liquid chromatography for rapid detection and identification of seven Candida species.

A novel denaturing high-performance liquid chromatography (DHPLC)-based technique allows rapid high-resolution analysis of PCR products. We used this technique for unequivocal molecular identification of seven Candida species. We show the application of this PCR/DHPLC approach for direct detection and identification of yeast species from blood cultures and for detection of Candida colonization in the gastrointestinal tract of allogeneic transplant patients.

Automated high-throughput mycobacterial interspersed repetitive unit typing of Mycobacterium tuberculosis strains by a combination of PCR and nondenaturing high-performance liquid chromatography.

Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing of Mycobacterium tuberculosis complex isolates is portable, 100% reproducible, and highly discriminatory. Nondenaturing high-performance liquid chromatography (non-dHPLC) with use of a WAVE microbial analysis system is a promising method of PCR amplicon analysis as it is low cost and requires no preanalysis processing. The aims of this study were to validate the application of WAVE microbial analysis system technology to MIRU-VNTR typing. A collection of 70 strains were cultivated in liquid culture and extracted using the QIAamp DNA minikit. Novel primers were designed to target the 12 MIRU-VNTR loci (P. Supply et al., J. Clin. Microbiol. 39:3563-3571, 2001). After amplification, each PCR product was analyzed on a WAVE microbial analysis system. The fragment size was calculated from the chromatogram, and the number of tandem repeats at each locus was determined. For the collection of 70 strains 100% concordance was achieved when comparing MIRU-VNTR profiles obtained from agarose gel electrophoresis and PCRs analyzed on a WAVE microbial analysis system. The calculated fragment sizes, obtained from the WAVE microbial analysis system, were sufficiently accurate to ensure 100% confidence when assigning the number of tandem repeats to a MIRU-VNTR locus. This study is the first to report the successful use of non-dHPLC for screening for variations in the number of MIRU-VNTRs in mycobacterial DNA. Non-dHPLC analysis was demonstrated to be a rapid, low-labor input method for the detection and analysis of MIRU-VNTR amplicons. The combination with non-dHPLC further enhances the utility of MIRU-VNTR typing.

Culture-independent identification of pathogenic bacteria and polymicrobial infections in the genitourinary tract of renal transplant recipients.

Renal transplant recipients are predisposed to urinary tract infections caused by both common uropathogens and opportunistic bacteria resulting frequently in significant polymicrobial infections. In this study, a culture-independent 16S rRNA-based approach was established to identify unusual, fastidious, or anaerobic bacteria and to investigate bacterial diversity in urinary tract specimens. Similarly sized amplicons encompassing the V6 to V8 region of the 16S rRNA were analyzed with denaturing high-performance liquid chromatography (DHPLC) (WAVE System). Artificial mixtures of single amplicons from commonly encountered uropathogenic bacteria produced distinct peak profiles whose identities were confirmed by sequencing individually collected peak products. We evaluated the application of the method on 109 urinary tract specimens from renal transplant recipients; 100% correlation was found for culture-positive specimens, and DHPLC generated peak profiles. However, for culture-negative specimens, DHPLC facilitated the detection of novel peak profiles. DNA sequencing of these individual peaks was used to identify the bacteria involved. Thus, in PCR-positive but culture-negative samples the method allowed detection of previously known uropathogens such as Corynebacterium urealyticum and Gardnerella vaginalis, but also unusual agents including Anaerococcus lactolyticus, Bacteroides vulgatus, Dialister invisus, Fusobacterium nucleatum, Lactobacillus iners, Leptotrichia amnionii, Prevotella buccalis, Prevotella ruminicola, Rahnella aquatilis, and Streptococcus intermedius were detected as single pathogens or as constituents of polymicrobial infections. The method described is reproducible and rapidly and enables both DHPLC-based profiling and sequence-based investigation of microbial communities and polymicrobial infections. A detailed understanding of infections found in recipients of renal transplants will guide antibiotic therapy regimens and provide new perspectives for decreasing the risk of graft rejection.

Endonuclease cleavage of blocked replication forks: An indirect pathway of DNA damage from antitumor drug-topoisomerase complexes.

The cytotoxicity of several important antitumor drugs depends on formation of the covalent topoisomerase-DNA cleavage complex. However, cellular processes such as DNA replication are necessary to convert the cleavage complex into a cytotoxic lesion, but the molecular mechanism of this conversion and the precise nature of the cytotoxic lesion are unknown. Using a bacteriophage T4 model system, we have previously shown that antitumor drug-induced cleavage complexes block replication forks in vivo. In this report, we show that these blocked forks can be cleaved by T4 endonuclease VII to create overt DNA breaks. The accumulation of blocked forks increased in endonuclease VII-deficient infections, suggesting that endonuclease cleavage contributes to fork processing in vivo. Furthermore, purified endonuclease VII cleaved the blocked forks in vitro close to the branch points. These results suggest that an indirect pathway of branched-DNA cleavage contributes to the cytotoxicity of antitumor drugs that target DNA topoisomerases.