Sex identification of ancient DNA samples using a microfluidic device.

Ancient DNA is the name given to the degraded, fragmented, and chemically damaged biomolecules that can be recovered from archaeological remains of plants, animals, and humans. Where ancient human DNA has survived at archaeological sites, it can give valuable information and is especially useful for its potential to identify kinship, population affinities, pathogens, and biological sex. Here, we describe the operation of a microfluidic device for the sex identification of ancient DNA samples using an efficient sample handling process. DNA is extracted from powdered bone samples and abasic sites labeled with biotin. Streptavidin-coated superparamagnetic particles are used to isolate the labeled DNA prior to amplification of the Amelogenin sex marker

The preparation of microfluidic architecture with monolithic materials using a dual porous silica structure.

A microfluidic device (MD) has been developed which features a porous silica (PS) monolithic disk synthesized from tetramethyl orthosilicate, incorporated into the device post-fabrication and sealed in place with a second PS monolithic layer, synthesized from potassium silicate. This dual porous silica (DPS) structure provides a pathway for sample introduction to the MD and offers an ideal platform for solid phase extraction (SPE) methodologies which can be rapidly and efficiently integrated into a chip-based format. All silica disk manufacture and functionalization was carried out in batch to provide a readily scalable method of production. Application of this design for processing samples was demonstrated using two alternative nucleic acid purification chemistries, yielding polymerase chain reaction amplifiable DNA extracted from 150 μL of human urine in less than 35 min. It is proposed that this DPS system could be further developed for a diverse range of chip-based SPE applications, providing an interface facilitating sample delivery and enabling SPE on-chip. Furthermore, to the author's knowledge it is the first reporting of two different types of PS amalgamated in a single MD

Visuo-tactile integration in autism: atypical temporal binding may underlie greater reliance on proprioceptive information

BackgroundEvidence indicates that social functioning deficits and sensory sensitivities in autism spectrum disorder (ASD) are related to atypical sensory integration. The exact mechanisms underlying these integration difficulties are unknown; however, two leading accounts are (1) an over-reliance on proprioception and (2) atypical visuo-tactile temporal binding. We directly tested these theories by selectively manipulating proprioceptive alignment and visuo-tactile synchrony to assess the extent that these impact upon body ownership.MethodsChildren with ASD and typically developing controls placed their hand into a multisensory illusion apparatus, which presented two, identical live video images of their own hand in the same plane as their actual hand. One virtual hand was aligned proprioceptively with the actual hand (the veridical hand), and the other was displaced to the left or right. While a brushstroke was applied to the participants’ actual (hidden) hand, they observed the two virtual images of their hand also being stroked and were asked to identify their real hand. During brushing, one of three different temporal delays was applied to either the displaced hand or the veridical hand. Thus, only one virtual hand had synchronous visuo-tactile inputs.ResultsResults showed that visuo-tactile synchrony overrides incongruent proprioceptive inputs in typically developing children but not in autistic children. Evidence for both temporally extended visuo-tactile binding and a greater reliance on proprioception are discussed.ConclusionsThis is the first study to provide definitive evidence for temporally extended visuo-tactile binding in ASD. This may result in reduced processing of amodal inputs (i.e. temporal synchrony) over modal-specific information (i.e. proprioception). This would likely lead to failures in appropriately binding information from related events, which would impact upon sensitivity to sensory stimuli, body representation and social processes such as empathy and imitation

Review of methods used by chiropractors to determine the site for applying manipulation

Background: With the development of increasing evidence for the use of manipulation in the management of musculoskeletal conditions, there is growing interest in identifying the appropriate indications for care. Recently, attempts have been made to develop clinical prediction rules, however the validity of these clinical prediction rules remains unclear and their impact on care delivery has yet to be established. The current study was designed to evaluate the literature on the validity and reliability of the more common methods used by doctors of chiropractic to inform the choice of the site at which to apply spinal manipulation. Methods: Structured searches were conducted in Medline, PubMed, CINAHL and ICL, supported by hand searches of archives, to identify studies of the diagnostic reliability and validity of common methods used to identify the site of treatment application. To be included, studies were to present original data from studies of human subjects and be designed to address the region or location of care delivery. Only English language manuscripts from peer-reviewed journals were included. The quality of evidence was ranked using QUADAS for validity and QAREL for reliability, as appropriate. Data were extracted and synthesized, and were evaluated in terms of strength of evidence and the degree to which the evidence was favourable for clinical use of the method under investigation. Results: A total of 2594 titles were screened from which 201 articles met all inclusion criteria. The spectrum of manuscript quality was quite broad, as was the degree to which the evidence favoured clinical application of the diagnostic methods reviewed. The most convincing favourable evidence was for methods which confirmed or provoked pain at a specific spinal segmental level or region. There was also high quality evidence supporting the use, with limitations, of static and motion palpation, and measures of leg length inequality. Evidence of mixed quality supported the use, with limitations, of postural evaluation. The evidence was unclear on the applicability of measures of stiffness and the use of spinal x-rays. The evidence was of mixed quality, but unfavourable for the use of manual muscle testing, skin conductance, surface electromyography and skin temperature measurement. Conclusions: A considerable range of methods is in use for determining where in the spine to administer spinal manipulation. The currently published evidence falls across a spectrum ranging from strongly favourable to strongly unfavourable in regard to using these methods. In general, the stronger and more favourable evidence is for those procedures which take a direct measure of the presumptive site of care– methods involving pain provocation upon palpation or localized tissue examination. Procedures which involve some indirect assessment for identifying the manipulable lesion of the spine–such as skin conductance or thermography–tend not to be supported by the available evidence.https://doi.org/10.1186/2045-709X-21-3

Antibacterial effects of theaflavin and synergy with epicatechin against clinical isolates of Acinetobacter baumannii and Stenotrophomonas maltophilia

Stenotrophomonas maltophilia and Acinetobacter baumannii are recognised as important nosocomial pathogens; however, due to their intrinsic resistance to multiple antibiotics, treatment options are limited. Polyphenols from black tea have been shown to possess antibacterial action. In this study, the antibacterial effects of various concentrations of theaflavin as well as combinations of theaflavin and epicatechin were determined using the disk diffusion assay. The results showed strong antibacterial activity of theaflavin against eight clinical isolates of S. maltophilia and A. baumannii. Significant synergy (P≤0.05) was also observed between theaflavin and epicatechin against all isolates. Although the mechanisms for this activity and synergy are not well understood, the clinical potential is clear and further research is recommended to determine the modes of action

Antifungal synergy of theaflavin and epicatechin combinations against Candida albicans

New antifungal agents are required to compensate for the increase in resistance to standard antifungal agents of C. albicans, which is an important opportunistic fungal pathogen that causes minor infections in many individuals but very serious infections in those who are immune-compromised. In this study, combinations of the theaflavin and epicatechin are investigated as potential antifungal agents and also to establish whether antifungal synergy exists between these two readily accessible and cost-effective polyphenols isolated from black and green tea. The results of disc diffusion assays show stronger antibacterial activity of theaflavin:epicatechin combinations against C. albicans NCTC 3255 and NCTC 3179, than that of theaflavin alone. Minimum inhibitory concentrations (MICs) of theaflavin and the theaflavin:epicatechin combinations of 1024 ㎍/mL with theaflavin and 128-256 ㎍/mL with theaflavin:epicatechin combinations are found. FICIs were calculated and synergy between theaflavin and epicatechin against both isolates of C. albicans confirmed. Theaflavin:epicatechin combinations show real potential for future use as a treatment for infections caused by C. albican

Minimum inhibitory and bactericidal concentrations of theaflavin and synergistic combinations with epicatechin and quercetin against clinical isolates of Stenotrophomonas maltophilia

Stenotrophomonas maltophilia is an important nosocomial pathogen with intrinsic resistance to multiple antibiotics. Previous investigations have shown flavanols from black tea to possess antibacterial activity. This study describes the determination of minimum inhibitory concentrations and minimum bactericidal concentration for theaflavin independently and in formulations with the polyphenols epicatechin and quercetin against nine clinical isolates of Stenotrophomonas maltophilia and the control isolate NCTC 130141 via the microtitre assay. The results demonstrate that theaflavin has strong antibacterial activity and also shows significant synergism with epicatechin and quercetin. The minimum inhibitory concentrations of the isolates range between 200-400 ug/mL for theaflavin and 100-200 ug/mL for both theaflavin:epicatechin and theaflavin:quercetin combinations. The minimum bactericidal concentrations were discovered to be a 2 fold increase on those of the minimum inhibitory concentrations. The research highlights the potential use of polyphenols for the clinical treatment of highly antibiotic resistant bacteria

Synergistic antibacterial effects of theaflavin in combination with ampicillin against hospital isolates of Stenotrophomonas maltophilia

Stenotrophomonas maltophilia is an important opportunistic nosocomial pathogen that shows intrinsic resistance to many antibiotics. This often limits treatment options and can cause lengthy hospital stays. Combination treatments are often used to combat resistance and using natural compounds such as polyphenols could give increased treatment options and even the reuse of antibiotics to which high levels of resistance have been observed. A checkerboard assay was used to determine if any synergy exists between ampicillin and the polyphenol theaflavin against 9 clinical isolates and one control isolate (NCTC 13014) of S. maltophilia. It was discovered that significant synergy (P 0.05) does exist between theaflavin and ampicillin, reducing the mean MIC of ampicillin from 12.5-22.9 μg/mL in liquid culture, to 3.125-6.25 μg/mL. The FIC index was calculated to be 0.22-0.35 confirming synergy. From these results, significant potential for medical applications can be seen and further investigation is recommended