Controlled placement of multiple CNS cell populations to create complex neuronal cultures.

In vitro brain-on-a-chip platforms hold promise in many areas including: drug discovery, evaluating effects of toxicants and pathogens, and disease modelling. A more accurate recapitulation of the intricate organization of the brain in vivo may require a complex in vitro system including organization of multiple neuronal cell types in an anatomically-relevant manner. Most approaches for compartmentalizing or segregating multiple cell types on microfabricated substrates use either permanent physical surface features or chemical surface functionalization. This study describes a removable insert that successfully deposits neurons from different brain areas onto discrete regions of a microelectrode array (MEA) surface, achieving a separation distance of 100 µm. The regional seeding area on the substrate is significantly smaller than current platforms using comparable placement methods. The non-permanent barrier between cell populations allows the cells to remain localized and attach to the substrate while the insert is in place and interact with neighboring regions after removal. The insert was used to simultaneously seed primary rodent hippocampal and cortical neurons onto MEAs. These cells retained their morphology, viability, and function after seeding through the cell insert through 28 days in vitro (DIV). Co-cultures of the two neuron types developed processes and formed integrated networks between the different MEA regions. Electrophysiological data demonstrated characteristic bursting features and waveform shapes that were consistent for each neuron type in both mono- and co-culture. Additionally, hippocampal cells co-cultured with cortical neurons showed an increase in within-burst firing rate (p = 0.013) and percent spikes in bursts (p = 0.002), changes that imply communication exists between the two cell types in co-culture. The cell seeding insert described in this work is a simple but effective method of separating distinct neuronal populations on microfabricated devices, and offers a unique approach to developing the types of complex in vitro cellular environments required for anatomically-relevant brain-on-a-chip devices.

Long-term non-invasive interrogation of human dorsal root ganglion neuronal cultures on an integrated microfluidic multielectrode array platform.

Scientific studies in drug development and toxicology rely heavily on animal models, which often inaccurately predict the true response for human exposure. This may lead to unanticipated adverse effects or misidentified risks that result in, for example, drug candidate elimination. The utilization of human cells and tissues for in vitro physiological platforms has become a growing area of interest to bridge this gap and to more accurately predict human responses to drugs and toxins. The effects of new drugs and toxins on the peripheral nervous system are often investigated with neurons isolated from dorsal root ganglia (DRG), typically with one-time measurement techniques such as patch clamping. Here, we report the use of our multi-electrode array (MEA) platform for long-term noninvasive assessment of human DRG cell health and function. In this study, we acquired simultaneous optical and electrophysiological measurements from primary human DRG neurons upon chemical stimulation repeatedly through day in vitro (DIV) 23. Distinct chemical signatures were noted for the cellular responses evoked by each chemical stimulus. Additionally, the cell viability and function of the human DRG neurons were consistent through DIV 23. To the best of our knowledge, this is the first report on long-term measurements of the cell health and function of human DRG neurons on a MEA platform. Future generations will include higher electrode numbers in customized arrangements as well as integration with different tissue types on a single device. This platform will provide a valuable testing tool for both rodent and human cells, enabling a more comprehensive risk assessment for drug candidates and toxicants.

High-temperature electromagnons in the magnetically induced multiferroic cupric oxide driven by intersublattice exchange.

Magnetically induced ferroelectric multiferroics present an exciting new paradigm in the design of multifunctional materials, by intimately coupling magnetic and polar order. Magnetoelectricity creates a novel quasiparticle excitation--the electromagnon--at terahertz frequencies, with spectral signatures that unveil important spin interactions. To date, electromagnons have been discovered at low temperature (<70 K) and predominantly in rare-earth compounds such as RMnO3. Here we demonstrate using terahertz time-domain spectroscopy that intersublattice exchange in the improper multiferroic cupric oxide (CuO) creates electromagnons at substantially elevated temperatures (213-230 K). Dynamic magnetoelectric coupling can therefore be achieved in materials, such as CuO, that exhibit minimal static cross-coupling. The electromagnon strength and energy track the static polarization, highlighting the importance of the underlying cycloidal spin structure. Polarized neutron scattering and terahertz spectroscopy identify a magnon in the antiferromagnetic ground state, with a temperature dependence that suggests a significant role for biquadratic exchange.

On-chip laser-induced DNA dehybridization.

Detection of pathogens and relevant genetic markers using their nucleic acid signatures is extremely common due to the inherent specificity genomic sequences provide. One approach for assaying a sample simultaneously for many different targets is the DNA microarray, which consists of several million short nucleic acid sequences (probes) bound to an inexpensive transparent substrate. Typically, complex samples hybridize to the microarray and the pattern of fluorescing probes on the microarray's surface identifies the detected targets. In the case of evolving or newly emergent organisms, a hybridization pattern can occur that differs from any previously known sources. When this happens it can be useful to recover the hybridized DNA from the binding locations of interest for sequencing. Here we present the novel utilization of a focused Infrared (IR) laser to heat user-selected spots on the DNA microarray surface, causing only localized dehybridization and recovery of the desired DNA into an elution buffer where it is available for subsequent amplification or sequencing. The introduction of a focused dehybridization method for spots of interest suppresses the amount of background DNA to be analyzed from downstream processes, and should reduce subsequent sequence assembly errors. This technique could also be applied to high-density protein microarrays where the desire to locally heat spots for release of bound molecules is desired.

Hypothalamic-pituitary-adrenal (HPA) axis activity in adults with intellectual disabilities: a preliminary investigation.

BACKGROUND: Cortisol is a marker of physiological arousal, exhibiting a characteristic pattern of diurnal activity. The daily cortisol profile has been xamined extensively and is atypical in a number of clinical disorders. However, there are very few studies focussing on the cortisol profile in adults with intellectual disabilities (ID). This paper reports a preliminary investigation into the nature of the cortisol profile in adults with mild or moderate ID and provides reflections on the challenges of psychophysiological research in this population. METHODS: On two consecutive days, 39 adults with mild or moderate ID each donated saliva samples for cortisol analysis, at multiple times between waking and evening. A comparison between these data and the published literature permitted a descriptive assessment of the cortisol awakening response (CAR) and diurnal profile. A variety of psychometric measures and an assessment of behavioural history were also collected in order to describe aspects of the participants' emotional and behavioural states. RESULTS: Individuals with ID exhibit a diurnal cortisol secretion profile, qualitatively similar to that of the typical, healthy, adult population. However, the findings also suggested a blunted CAR, warranting further investigation. There was also some evidence that cortisol secretion was affected by anxiety and a recent history of aggression. CONCLUSION: While further work is required to characterise the CAR fully, there was no indication that the diurnal cortisol profile among people with ID differs from that of the typical population. This study also demonstrates that, although challenging, it is feasible, and acceptable to participants, to collect repeated physiological measures from men and women with mild and moderate ID.

Asymmetric thermal line shape broadening in a gapped 3D antiferromagnet: evidence for strong correlations at finite temperature.

It is widely believed that magnetic excitations become increasingly incoherent as the temperature is raised due to random collisions which limit their lifetime. This picture is based on spin-wave calculations for gapless magnets in 2 and 3 dimensions and is observed experimentally as a symmetric Lorentzian broadening in energy. Here, we investigate a three-dimensional dimer antiferromagnet and find unexpectedly that the broadening is asymmetric-indicating that far from thermal decoherence, the excitations behave collectively like a strongly correlated gas. This result suggests that a temperature activated coherent state of quasiparticles is not confined to special cases like the highly dimerized spin-1/2 chain but is found generally in dimerized antiferromagnets of all dimensionalities and perhaps gapped magnets in general.

HIV and dentistry in Australia: clinical and legal issues impacting on dental care.

The number of people in Australia living with HIV is growing. This reflects a consistent rate of new HIV infections combined with an increased life expectancy of people with HIV. Dentists are ideally positioned to identify, manage and treat HIV-associated oral manifestations and have a responsibility to themselves and to their patients to be up-to-date with the evolving area of HIV and related issues. Those issues include medico-legal implications associated with HIV diagnosis and treatment. This article provides a review of the current clinical and medico-legal aspects of HIV in Australia. The oral manifestations of HIV can be divided into five categories: microbiological infections (fungal, bacterial and viral); oral neoplasms; neurological conditions; other oral conditions that may be associated with HIV infection; and oral conditions associated with HIV treatment. Current treatment options in the scope of general dental practice are outlined. Medico-legal issues related to the management of patients with HIV are explored, including rights of the patient regarding disclosure of HIV status; an algorithm for the management of a patient with signs or symptoms indicating possible HIV infection, including referral pathways; and an algorithm for dealing with patient management and referral issues.

Under-three minute PCR: probing the limits of fast amplification.

Nucleic acid amplification is enormously useful to the biotechnology and clinical diagnostic communities; however, to date point-of-use PCR has been hindered by thermal cycling architectures and protocols that do not allow for near-instantaneous results. In this work we demonstrate PCR amplification of synthetic SARS respiratory pathogenic targets and bacterial genomic DNA in less than three minutes in a hardware configuration utilizing convenient sample loading and disposal. Instead of sample miniaturization techniques, near-instantaneous heating and cooling of 5 µL reaction volumes is enabled by convective heat transfer of a thermal fluid through porous media combined with an integrated electrical heater. This method of rapid heat transfer has enabled 30 cycles of PCR amplification to be completed in as little as two minutes and eighteen seconds. Surprisingly, multiple enzymes have been shown to work at these breakthrough speeds on our system. A tool for measuring enzyme kinetics now exists and can allow polymerase optimization through directed evolution studies. Pairing this instrument technology with modified polymerases should result in a new paradigm for high-throughput, ultra-fast PCR and will hopefully improve our ability to quickly respond to the next viral pandemic.

Field olfactometry assessment of dairy manure land application methods.

Surface application of manure in reduced tillage systems generates nuisance odors, but their management is hindered by a lack of standardized field quantification methods. An investigation was undertaken to evaluate odor emissions associated with various technologies that incorporate manure with minimal soil disturbance. Dairy manure slurry was applied by five methods in a 3.5-m swath to grassland in 61-m-inside-diameter rings. Nasal Ranger Field Olfactometer (NRO) instruments were used to collect dilutions-to-threshold (D/T) observations from the center of each ring using a panel of four odor assessors taking four readings each over a 10-min period. The Best Estimate Threshold D/T (BET10) was calculated for each application method and an untreated control based on preapplication and <1 h, 2 to 4 h, and approximately 24 h after spreading. Whole-air samples were simultaneously collected for laboratory dynamic olfactometer evaluation using the triangular forced-choice (TFC) method. The BET10 of NRO data composited for all measurement times showed D/T decreased in the following order (a = 0.05): surface broadcast > aeration infiltration > surface + chisel incorporation > direct ground injection Sshallow disk injection > control, which closely followed laboratory TFC odor panel results (r = 0.83). At 24 h, odor reduction benefits relative to broadcasting persisted for all methods except aeration infiltration, and odors associated with direct ground injection were not different from the untreated control. Shallow disk injection provided substantial odor reduction with familiar toolbar equipment that is well adapted to regional soil conditions and conservation tillage operations.

Quantum criticality in an Ising chain: experimental evidence for emergent E8 symmetry.

Quantum phase transitions take place between distinct phases of matter at zero temperature. Near the transition point, exotic quantum symmetries can emerge that govern the excitation spectrum of the system. A symmetry described by the E8 Lie group with a spectrum of eight particles was long predicted to appear near the critical point of an Ising chain. We realize this system experimentally by using strong transverse magnetic fields to tune the quasi-one-dimensional Ising ferromagnet CoNb2O6 (cobalt niobate) through its critical point. Spin excitations are observed to change character from pairs of kinks in the ordered phase to spin-flips in the paramagnetic phase. Just below the critical field, the spin dynamics shows a fine structure with two sharp modes at low energies, in a ratio that approaches the golden mean predicted for the first two meson particles of the E8 spectrum. Our results demonstrate the power of symmetry to describe complex quantum behaviors.

Effect of infrared temperature on thermoregulatory behaviour in suckling piglets.

The objective of this study was to investigate the effect of infrared (IR) temperature on thermoregulatory behaviour in suckling piglets in the first 3 weeks after farrowing. A total of 10 piglets from each of the 16 litters were exposed to recommended IR temperature conditions at 1, 2 and 3 weeks of age, with a mild offset (4°C) in IR temperature during the first experiment and a more challenging offset (8°C) during the second experiment. Digital photos were taken when all piglets had settled in the creep area, and the lying posture and huddling behaviour were analysed. A lying posture score and a huddling score was calculated by multiplying the number of piglets in each category with a given value for each category, based on different lying postures and different degrees of huddling behaviour. With a 4°C change in IR temperature, the piglets tended to alter their lying posture, while an 8°C change had a significant effect on lying posture (P < 0.01). A change in IR temperature of 4°C had no effect on the degree of huddling. The huddling score decreased significantly with 8°C change in IR temperature (P < 0.05). Postural changes, rather than changes in degree of huddling were the preferred thermoregulatory strategy for suckling piglets.

Short communication: effect of changing the ratio of forage to concentrate on ammonia emissions by dairy heifers.

Two animal growth studies and a companion digestibility study were conducted to evaluate the effect of differing ratios of forage to concentrate and the addition of yeast culture (Saccharomyces cerevisiae) on NH(3) emissions from the manure of growing dairy heifers with corn silage (CS) as the sole forage. Flux chamber methods were used to measure NH(3) volatilization from the barn floor or by laboratory procedures. In experiment 1, 24 Holstein heifers (159 +/- 3.3 kg of initial body weight; BW) were fed either a low-concentrate diet (LC; 77% CS, 23% concentrate) or a high-concentrate diet (HC; 33% CS, 67% concentrate) in a randomized design. Manure (feces and urine mixture) from heifers consuming the LC diets volatilized similar amounts of NH(3) as manure from HC heifers (314.0 vs. 174.4 +/- 36.1 microg/cm(2) per min). In experiment 2, 24 older heifers (227.9 +/- 27.1 kg of BW) were used. Manure from HC heifers released slightly less NH(3) from the barn floor, confirming the results from the initial study. Finally, a digestibility study was undertaken using four 9-mo-old heifers (234 +/- 15 kg of initial BW) and four 14-mo-old heifers (409 +/- 20 kg of initial BW), allocated to 4 treatments consisting of an HC or LC diet with or without yeast culture addition. Emissions per unit of manure (mg of NH(3)/g) from heifers in both age groups were greater for the HC diets; however, total emissions per day were equal. Yeast culture addition had no effect on cumulative daily emissions. In these 3 experiments, NH(3) emissions from HC heifers were not different from those from LC heifers.

Orbital degeneracy removed by charge order in triangular antiferromagnet AgNiO2.

We report a high-resolution neutron diffraction study on the orbitally degenerate spin-1/2 hexagonal metallic antiferromagnet AgNiO2. A structural transition to a tripled unit cell with expanded and contracted NiO6 octahedra indicates sqrt[3]xsqrt[3] charge order on the Ni triangular lattice. This suggests charge order as a possible mechanism of lifting the orbital degeneracy in the presence of charge fluctuations, as an alternative to the more usual Jahn-Teller distortions. A novel magnetic ground state is observed at low temperatures with the electron-rich S=1 Ni sites arranged in alternating ferromagnetic rows on a triangular lattice, surrounded by a honeycomb network of nonmagnetic and metallic Ni ions. We also report first-principles band-structure calculations that explain microscopically the origin of these phenomena.

Differences in stem and progenitor cell yield in different subcutaneous adipose tissue depots.

BACKGROUND: Human adipose tissue has been shown to contain multipotent cells with properties similar to mesenchymal stromal cells. While there have been many studies of the biology of these cells, no study has yet evaluated issues associated with tissue harvest. METHODS: Adipose tissue was obtained from the subcutaneous space of the abdomen and hips of 10 donors using both syringe and pump-assisted liposuction. Tissue was digested with collagenase and then assayed for the presence of different stem and progenitor cell types using clonogenic culture assays, including fibroblast colony-forming unit (CFU-F) and alkaline phosphatase-positive colony-forming unit (CFU-AP). Paired analysis of samples obtained from the same individual was used to compare harvest method and site. RESULTS: Syringe suction provided significantly greater recovery of adipocytes and a non-significant trend towards improved recovery of cells in the adipocyte-depleted fraction. There was considerable donor-to-donor variation in stem cell recovery. However, paired analysis of tissue obtained from different subcutaneous sites in the same donor showed that tissue harvested from the hip yielded 2.3-fold more CFU-F/unit volume and a 7-fold higher frequency of CFU-AP than that obtained from the abdomen. These differences were statistically significant. DISCUSSION: Harvest site influences the stem and progenitor cell content of subcutaneous adipose tissue.

Genome-wide scan for loci influencing quantitative immune response traits in the Belém family study: comparison of methods and summary of results.

Here we report the results from a genome-wide linkage scan to identify genes and chromosomal regions that influence quantitative immune response traits, using multi-case leprosy and tuberculosis families from north-eastern Brazil. Total plasma IgE, antigen-specific IgG to Mycobacterium leprae soluble antigen (MLSA), M. tuberculosis soluble antigen (MTSA) and M. tuberculosis purified protein derivative (PPD), and antigen-specific lymphocyte proliferation (stimulation index or SI) and interferon-gamma (IFN-gamma) release to MLSA and PPD, were measured in 16 tuberculosis (184 individuals) and 21 leprosy (177 individuals) families. The individuals were genotyped at 382 autosomal microsatellite markers across the genome. The adjusted immune-response phenotypes were analysed using a variety of variance components and regression-based methods. These analyses highlighted a number of practical issues and problems with regard to implementation of the methods and, interestingly, differences were observed between several standard statistical and genetic analysis packages used. From this we determined that, for this set of traits in these pedigrees, significant p values for linkage using variance components analysis, supported by significance using the Visscher-Hopper modification of the Haseman-Elston method, provided the most compelling evidence for linkage. Using these criteria, linkage (5.8 x 10(-5) < p < 0.008) was seen for: total plasma IgE on chromosome 2; IgG to MLSA on chromosomes 8, 17 and 21; IgG to PPD on chromosome 12; SI to PPD on chromosome 1; IFN-gamma to MLSA on chromosomes 6, 7, 10, 12 and 14; and IFN-gamma to PPD on chromosomes 1, 16 and 19.

Brain-derived neurotrophic factor is expressed in a gradient in the superior colliculus during development of the retinocollicular projection.

Abstract Theoretical models of topographic map formation have postulated a gradient of attractant in addition to a gradient of repulsion in the target. In species where many axons grow past their correct positions initially, it has also been argued that a parallel gradient of attractant or branching signal is required to ensure collateral formation at the correct position (O'Leary et al., 1999). Brain-derived neurotrophic factor (BDNF) is a known attractant and promotes branching of retinal axons. We have examined its distribution in the superior colliculus and that of its receptor, trkB, in the retina, using immunohistochemistry and in situ hybridization, respectively, during the development of the topographic retinocollicular projection in the wallaby, a marsupial mammal. The number of glial endfeet expressing BDNF at the surface of the colliculus was found to be in a high caudal-to-low rostral gradient during the time when the retinocollicular projection was developing. When the projection was mature the rostrocaudal gradient had disappeared and the number of detectable endfeet expressing BDNF was very low. Messenger RNA for TrkB was expressed in the retinal ganglion cell layer throughout the time when the retinocollicular projection was developing, with no difference in expression across the nasotemporal axis of the retina. The low rostral to high caudal distribution of BDNF in glial endfeet supports the idea that it is providing a parallel gradient of attractant or branching signal in the colliculus.

Convectively driven polymerase chain reaction thermal cycler.

We have fabricated a low-cost disposable polymerase chain reaction thermal chamber that uses buoyancy forces to move the sample solution between the different temperatures necessary for amplification. Three-dimensional, unsteady finite element modeling and a simpler 1-D steady-state model are used together with digital particle image velocimetry data to characterize the flow within the device. Biological samples have been amplified using this novel thermal chamber. Time for amplification is less than 30 min. More importantly, an analysis of the energy consumption shows significant improvements over current technology.

Incidence of nondextrality in the NAS/NRC Twin Registry.

Genetic theories still flounder on the fact that similarity of hand preference is the same in monozygotic (MZ) and dizygotic (DZ) twins. Lateral preference on a well-designed set of 5 activities was obtained from 2,131 male pairs. On item analysis, only "throw" discriminated zygosity, attributable to "excess" nondextral MZ pairs. This item is remarkably free of the intense cultural bias against sinistrality.

The National Health Service Breast Screening Programme and British Association of Surgical Oncology audit of quality assurance in breast screening 1996-2001.

BACKGROUND: The National Health Service Breast Screening Programme (NHSBSP) is an example of a nationally coordinated quality assurance programme in which all the professional groups involved participate. METHODS: Surgeons, radiologists and pathologists defined the clinical outcome measures against which they would subsequently be audited. The NHSBSP and the Association of Breast Surgery at BASO are jointly responsible for coordinating an annual audit of all surgical activities undertaken within the NHSBSP. RESULTS: The trends for key outcome measures between 1996 and 2001 are provided. The preoperative diagnosis rate (minimum standard 70 per cent or more) improved from 63 to 87 per cent. This rise was mirrored by an increase in the use of core biopsy in preference to fine-needle cytology. The proportion of patients in whom lymph node status was recorded improved from 81 to 93 per cent. There was no significant change in the number of women treated by low case-load surgeons and waiting times for surgery increased through the study interval. CONCLUSION: The BASO-NHSBSP Breast Audit has recorded major changes in clinical practice over 5 years. A key feature has been the dissemination of good practice through feedback of the results at local and national level.