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Background Dementia is a leading cause of disease burden in Australia, with almost half a million people living with dementia and a steady increase over time in dementia-related deaths. Strengthening the evidence base for dementia risk factors is critical for an effective and efficient public health and policy response. However, the number of Australians with dementia remains unknown, and there are significant gaps in the knowledge on risk factors in the Australian context. In this study we aim to develop reliable data on dementia incidence in Australia and investigate the associated risk factors, using a large population cohort. Specifically, we will assess the relative contribution of risk factors to dementia incidence as a basis for strengthening dementia prevention efforts. Methods We will use data from the 45 and Up Study that includes 267,358 residents of New South Wales, Australia, aged over 45 years, recruited between 2006-2009. To identify dementia cases we will link data from the 45 and Up Study with multiple health datasets containing information relevant to dementia case identification. We will estimate age- and sex-specific dementia incidence and model the association between dementia and risk factors related to socio-demographic characteristics, health conditions and health behaviours. We will also estimate the impact of various modifiable exposures on dementia incidence. Based on the results, we will produce a series of knowledge translation products providing advice on the contribution of identified risk factors to dementia incidence in Australia. Discussion Linking the 45 and Up Study data to multiple health datasets provides a unique opportunity to explore the role of risk factors on dementia incidence, including modelling the effect of modifiable risk factors on dementia incidence in the Australian population. We anticipate the results from this study to guide targeted and gradated strategies for population-level dementia prevention.

Background Peripheral caries may cause significant oral pain and pathology and is very prevalent within the Western Australia horse population. Associations with periodontal disease have been indicated; however, further work is needed to assess the anatomical and histological aspects of the conditions, to better understand the pathophysiology. Objectives To assess the anatomical and histopathological changes associated with equine cheek teeth peripheral caries and disease of the periodontium to better understand the pathogenesis and any association between the conditions. Study design Cross-sectional epidemiological and histological study. Methods A survey of 500 Western Australia horses was performed to assess the prevalence of peripheral caries and associations with other dental pathologies within the Western Australia horse population. Histopathological assessment was also performed on four extracted cheek teeth affected by peripheral caries and on three interdental areas from an abattoir specimen affected by peripheral caries and interdental feed accumulation. Results There was a significant association between peripheral caries and cheek teeth interdental feed accumulation and gingival recession. This association was significantly stronger in the mandibular cheek teeth than the maxillary cheek teeth and also in horses with moderate or severe peripheral caries compared to horses with mild peripheral caries. Histopathological examination found caries lesions consistent with those found in humans above the gingival margin. Sub-gingivally, however, the cementum and periodontal structures were normal. In the samples with concurrent peripheral caries and deep feed-pocketing, there was significant gingival recession; however, only mild or no histopathological changes occurred in the gingiva. Main limitations Small sample size for histopathological assessment. Conclusion Horses with peripheral caries, and in particular, horses with mandibular cheek teeth with peripheral caries are significantly more likely to also be affected by disease of the periodontium than horses without peripheral caries.

BACKGROUND Equine peripheral caries can cause significant morbidity and can have considerable welfare implications. Recent research suggests that diets with high water soluble carbohydrate (WSC) content are associated with high risk of peripheral caries. Previous work has indicated that the condition may be treatable if the inciting cause is removed and the damaged tooth allowed to erupt out, being replaced by the unaffected tooth previously under the gingival margin. OBJECTIVES To see whether the peripheral caries process can be ceased if oaten hay (typically high WSC) is removed from the diet and replaced with a non-cereal hay (typically lower WSC). STUDY DESIGN Retrospective blinded longitudinal study. METHODS Forty-two cases with peripheral caries that were on oaten hay were asked if they would change the hay type from oaten hay to a non-cereal hay or straw (meadow, Rhodes hay, lucerne hay or barley straw). Photographs were taken at the time and then again at subsequent visits. The photographs were anonymised, randomised and scored by six equine veterinary dentists using the Jackson et al. 'Peripheral Caries Grading System' grading scale. RESULTS At follow-up, 69.0% of cases were marked as inactive, compared with 47.6% of cases at baseline (OR: 2.45, 95%CI: 1.12-5.36, P = .02). Significantly lower grades of peripheral caries were observed in the gingival, middle and occlusive third of the molars (triadan 9-11's) at follow-up compared with baseline. However, significant improvements were not observed in the premolars (triadan 6-8's). MAIN LIMITATIONS This is a review of clinical records, not a prospective study. As such, other changes in the diet and management were not recorded, and there was no control group. CONCLUSIONS Recommending clients change their horse's diet from oaten hay (high WSC) to a typically lower WSC hay was associated with significant improvements in equine peripheral caries located in the molars.

Equine peripheral caries (PC) can cause significant dental pathology and appears to be increasing in prevalence and recognition in many areas [1, 2]. Previous studies have identified risk factors for the condition including the feeding of oaten hay [3]. It was hypothesized that this may be due to the higher water soluble carbohydrate (WSC) or “sugar” content of the hay. A randomized control trial involving 30 horses on three properties was completed. The horses were randomly assigned to two groups: high WSC (H-WSC) or low WSC (L-WSC) oaten hay and were then sedated and intraoral photographs and endoscopy were performed for baseline levels. They were maintained on this assigned hay source for three months when they were again sedated, examined, and photographs and endoscopic videos recorded. Horses with significant PC were then changed to meadow hay (previously shown to be lower risk for PC [3]) for seven months and re-examined to assess for recovery. Images and videos were anonymized and graded for PC, and the PC grades of the teeth before and after intervention were compared using a Fisher exact test. Thirteen horses fed L-WSC hay and 15 fed H-WSC hay completed the study. Of horses fed the H-WSC oaten hay, 60% deteriorated and of horses on L-WSC oaten hay, 53.8% improved in PC grade over the trial, however, the difference was not statistically significant (P = .274). All three horses available for recheck after changing to meadow hay improved.

BACKGROUND Equine peripheral caries can be a significant welfare issue in affected horses. Recent work has shown that complete reversal of the condition may be possible if the primary risk factor can be identified and corrected. Determining whether caries are active (deteriorating) or inactive (improving) is of vital importance when formulating a management and treatment plan. A new grading system is proposed that looks at whether caries are likely active or inactive to assist clinicians deciding whether intervention is required. OBJECTIVES To examine the reliability of an alternative grading method for equine peripheral caries. STUDY DESIGN Online survey of veterinary surgeons. METHODS Reviewers (n = 22) graded images of peripheral caries using three methods: the traditional grading system, a new research grading system and a new clinical grading system. Percentage agreement and Fleiss' kappa were used to examine the participant's agreement of the severity of caries. RESULTS All percentage agreements and Fleiss' kappa values were significantly greater than zero (P

BACKGROUND Peripheral dental caries is defined as macroscopic destruction of the calcified dental tissues and can cause considerable dental pathology and pain. It appears to occur at a high prevalence in Western Australian horses. At present, risk factors for the condition are poorly understood, making treatment and prevention difficult. OBJECTIVES To assess the prevalence of and potential risk factors for peripheral caries in Western Australian horses. STUDY DESIGN Cross-sectional, epidemiological study. METHODS A survey of 500 Western Australian horses was administered in two sections. The first section was completed by the owner and referred to the horse's signalment, diet and husbandry conditions. The second section was completed by veterinarians and focused on the horse's oral health. A multivariable logistic regression model was used to assess risk factors associated with peripheral caries. RESULTS Peripheral caries was present in 58.8% (95% confidence interval [CI] 54.5-63.1%) of surveyed horses. Breed was significantly associated with peripheral caries, with Warmbloods (odds ratio [OR] 0.44, 95% CI 0.24-0.82; P = 0.009) and Western breeds (OR 0.38, 95% CI 0.19-0.78; P = 0.008) being less likely to have peripheral caries than Thoroughbreds. Dietary risk factors included oaten hay (OR 2.90, 95% CI 1.87-4.53; P

Modern single-use microfluidic devices allow for affordable analysis of liquid, while remaining robust, easy-to-use and portable. Reliable and simple valving is crucial for fluid manipulation in these devices. Passive valves such as geometry-based burst valves are a frequent choice, allowing for instrumentation-free low-complexity valving. However, transfer from prototyping to manufacturing or implementation of changes in the manufacturing process oftentimes severely affect burst valve performance as material choice and surface topography impact the delicate balance of surface tension and liquid pressure in a channel. We, therefore, started this work with the goal of analyzing the influence of manufacturing techniques and materials on burst valve behavior. Yet, we soon came to learn about limitations of geometry-based burst valves when employed for commercially manufactured disposable microfluidic systems. Thence, this work presents an analytical prediction of burst valve pressure, compares predicted and experimental burst pressures based on common prototyping and manufacturing means and materials, and specifically analyses limitations of burst valves for commercial microfluidic devices. Factors influencing burst valve behavior are discussed and recommendations for robust implementation of burst valves are made.

BACKGROUND A large quantity of data is collected during the delivery of cancer care. However, once collected, these data are difficult for health professionals to access to support clinical decision making and performance review. There is a need for innovative tools that make clinical data more accessible to support health professionals in these activities. One approach for providing health professionals with access to clinical data is to create the infrastructure and interface for a clinical dashboard to make data accessible in a timely and relevant manner. OBJECTIVE This study aimed to develop and evaluate 2 prototype dashboards for displaying data on the identification and management of lymphedema. METHODS The study used a co-design framework to develop 2 prototype dashboards for use by health professionals delivering breast cancer care. The key feature of these dashboards was an approach for visualizing lymphedema patient cohort and individual patient data. This project began with 2 focus group sessions conducted with members of a breast cancer multidisciplinary team (n=33) and a breast cancer consumer (n=1) to establish clinically relevant and appropriate data for presentation and the visualization requirements for a dashboard. A series of fortnightly meetings over 6 months with an Advisory Committee (n=10) occurred to inform and refine the development of a static mock-up dashboard. This mock-up was then presented to representatives of the multidisciplinary team (n=3) to get preliminary feedback about the design and use of such dashboards. Feedback from these presentations was reviewed and used to inform the development of the interactive prototypes. A structured evaluation was conducted on the prototypes, using Think Aloud Protocol and semistructured interviews with representatives of the multidisciplinary team (n=5). RESULTS Lymphedema was selected as a clinically relevant area for the prototype dashboards. A qualitative evaluation is reported for 5 health professionals. These participants were selected from 3 specialties: surgery (n=1), radiation oncology (n=2), and occupational therapy (n=2). Participants were able to complete the majority of tasks on the dashboard. Semistructured interview themes were categorized into engagement or enthusiasm for the dashboard, user experience, and data quality and completeness. CONCLUSIONS Findings from this study constitute the first report of a co-design process for creating a lymphedema dashboard for breast cancer health professionals. Health professionals are interested in the use of data visualization tools to make routinely collected clinical data more accessible. To be used effectively, dashboards need to be reliable and sourced from accurate and comprehensive data sets. While the co-design process used to develop the visualization tool proved effective for designing an individual patient dashboard, the complexity and accessibility of the data required for a cohort dashboard remained a challenge.

Cell-free protein synthesis (CFPS), which entails synthesizing proteins outside of intact cells, is conducted in several formats with the continuous-exchange cell-free (CECF) format generally having the greatest protein expression yields. With this format, continuous chemical exchange occurs through a dialysis membrane separating a reaction solution from a feeding solution containing supplemental nutrient/energy molecules. Here, we describe the optimization of the miniaturized fluid array device (µFAD) by studying the effects of structural and experimental parameters responsible for the heightened chemical exchange across the dialysis membranes and enhanced protein expression capabilities of the high-throughput device. The interface area and number between the reaction and feeding solutions have a direct impact on protein expression, with a 1.6% enhancement in protein expression yield with each square millimeter increase in area and a 20% decrease with each additional interface. For nutrient/energy availability, an increasing solution volume ratio and height difference increase protein expression yield until the expression yield plateaus at a volume ratio of 20 to 1 (feeding to reaction solution) and a solution height difference of 2 mm. This yield can be further increased by 7% every 30 min with feeding solution replacement. Of the studied experimental factors (feeding solution stirring, device shaking, and temperature increase), feeding solution stirring has a significant effect on protein expression in this device. In the optimized system, green fluorescent protein (GFP), s-glucuronidase (GUS), s-galactosidase (LacZ), luciferase, and tissue plasminogen activator (tPA) expression increased 77.8-, 212-, 3.66-, 463-, and 5.43-fold, respectively, compared to the conventional batch format in a standard microplate. These results highlight the significance of structural/experimental conditions on the productive expression of proteins in the CECF format. Biotechnol. Bioeng. 2015;112: 2459–2467. © 2015 Wiley Periodicals, Inc.

To build replicating systems with new functions, the engineering of existing biological machineries requires a sensible strategy. Protein synthesis Using Recombinant Elements (PURE) system consists of the desired components for transcription, translation, aminoacylation and energy regeneration. PURE might be the basis for a radically alterable, lifelike system after optimization. Here, we regenerated 54 E. coli ribosomal (r-) proteins individually from DNA templates in the PURE system. We show that using stable isotope labeling with amino acids, mass spectrometry based quantitative proteomics could detect 26 of the 33 50S and 20 of the 21 30S subunit r-proteins when coexpressed in batch format PURE system. By optimizing DNA template concentrations and adapting a miniaturized Fluid Array Device with optimized feeding solution, we were able to cogenerate and detect at least 29 of the 33 50S and all of the 21 30S subunit r-proteins in one pot. The boost on yield of a single r-protein in coexpression pool varied from ∼1.5 to 5-fold compared to the batch mode, with up to ∼2.4 μM yield for a single r-protein. Reconstituted ribosomes under physiological condition from PURE system synthesized 30S r-proteins and native 16S rRNA showed ∼13% activity of native 70S ribosomes, which increased to 21% when supplemented with GroEL/ES. This work also points to what is still needed to obtain self-replicating synthetic ribosomes in situ in the PURE system.

Periodontal disease of equine cheek teeth is common and may lead to tooth loss if left untreated. Limited information is available comparing the effectiveness of treatment methods. The objective of this study was to retrospectively compare the effectiveness of 4 commonly used treatments in reducing periodontal pocket depth (in addition to routine dental treatment and occlusal equilibration). The 4 treatments compared were (1) removal of feed material, lavaging the pocket with dilute chlorhexidine, and then rinsing the mouth with chlorhexidine-containing mouthwash (CL); (2) CL plus placement of metronidazole into the pocket (M); (3) M plus the addition of polyvinyl siloxane temporary filling over the diastema (PVS); and (4) diastema widening to increase the interdental space, then PVS (DW). Pocket measurements were compared before and 2 to 6 months after treatment. Treatment groups CL, M, and PVS showed statistically significant reductions in pocket depth following treatment. The mean pocket depth reduction was the greatest in the DW group (and this was the only group with no cases having an increase in pocket depth), but this was not significant due to the small sample size. Additional analysis to compare effectiveness revealed a confounding effect of initial pocket depth. After accounting for this, DW was associated with smaller improvements than the other treatments, however, this was also based on a small sample size. After accounting for confounders, differences between treatments CL, M and PVS were not found to be significant, although all were associated with statistically significant reductions in pocket depth.

Reasons for performing the study. Equine cheek teeth periodontal disease is a common, often painful dental condition that may lead to premature tooth loss if left untreated. All cases of periodontal disease in this study were associated with diastemata, with the majority of cases being secondary or senile diastemata. At present limited objective information is available comparing different treatment methods for the condition to assist clinicians in making evidence-based treatment decisions. Objectives. To compare the effectiveness of four commonly used treatments for equine periodontal disease (additional to routine dental treatment and equilibration) to assess their merit in reducing periodontal pocket depth. Study design. Retrospective case series. Methods. Four commonly used equine periodontal treatments were compared: Removing feed material and lavaging the periodontal pocket with dilute chlorhexidine and rinsing the mouth with chlorhexidine based mouthwash (‘Hexarinse’), ‘clean and antiseptic lavage’ (CL); CL plus the addition of metronidazole antibiotics into the periodontal pockets (M); M plus the addition of a polyvinyl siloxane temporary filling over the diastema (PVS); and diastema widening to open the diastema and increase the interdental space between adjacent teeth, then PVS (DW). Periodontal pocket depth measurements were compared before and after treatment at treatment intervals between two and six months. Results. Treatment groups CL, M and PVS showed statistically significant reductions in pocket depth following treatment. Mean pocket depth reduction was greatest in the DW group (and this was the only group with no cases involving an increase in pocket depth), but this was based on a small sample size and was not statistically significant. Additional analysis to compare effectiveness revealed a strong confounding effect of initial pocket depth. After taking this into account, there was some evidence that DW was associated with smaller improvements than the other treatments, however this result was also based on a small sample size, and influenced by a small number of cases with particularly large improvements. Among the other treatments, no statistically significant differences in effectiveness were found. Conclusion. This study has shown that treatment methods CL, M and PVS are associated with statistically significant reductions in pocket depth. Due to the confounding effect of initial pocket depth, no clear differences in effectiveness were found between treatment methods.

Analysis of the intrinsic fluorescence profiles of individual marine algae can be used in general classification of organisms based on cell size and fluorescence properties. We describe the design and fabrication of a Microflow Cytometer on a chip for characterization of phytoplankton. The Microflow Cytometer measured distinct side scatter and fluorescence properties of Synechococcus sp., Nitzschia d., and Thalassiosira p.; measurements were confirmed using the benchtop Accuri C6 flow cytometer. The Microflow Cytometer proved sensitive enough to detect and characterize picoplankton with diameter approximately 1 μm and larger phytoplankton of up to 80 μm in length. The wide range in size discrimination coupled with detection of intrinsic fluorescent pigments suggests that this Microflow Cytometer will be able to distinguish different populations of phytoplankton on unmanned underwater vehicles.

Reasons for performing the study. Equine periodontal disease is a common, often painful dental condition that may lead to premature tooth loss if left untreated [1,2,3]. At present little research has been done comparing different treatment methods for the condition to assist clinicians in making educated treatment decisions. Objectives. To compare the effectiveness of four commonly used treatment options for periodontal disease (on top of routine dental treatment and equilibration) to assess their merit in reducing periodontal pocket depth. Study Design. Retrospective case series. Methods. Four commonly used treatment options were compared: Removing feed material and disinfecting the periodontal pocket with dilute chlorhexidine and rinsing the mouth with chlorhexidine based mouthwash (‘Hexarinse’), ‘cleaned and disinfected’ (CD); CD plus the addition of metronidazole antibiotics into the periodontal pockets (M); M plus the addition of a polyvinyl siloxane temporary filling over the diastema (PVS); and diastema burring to widen the gap between the teeth, then PVS (DB). Pocket depth measurements were compared before and after treatment at treatment intervals between two and six months. Results. Treatment groups CD, M and PVS showed statistically significant reductions in pocket depth between visits. Mean reduction was greatest in the DB group, but this was not statistically significant. Additional analysis to compare effectiveness revealed a strong confounding effect of initial pocket depth. After taking this into account, there was some evidence that DB was associated with smaller improvements than the other treatments, however this result was based on a small sample size. Among the other treatments, no statistically significant differences in effectiveness were found. Conclusion. This study has shown that treatment methods CD, M and PVS are associated with statistically significant reductions in pocket depth. Due to the confounding effect of initial pocket depth, no clear differences in effectiveness were found between treatment methods.

Compared to cell-based protein expression, cell-free protein synthesis (CFPS) offers several advantages including a greater control over system additives. This control is further enhanced with a CFPS system called the Protein synthesis Using Recombinant Elements (PURE) system, which consists of 108 purified transcriptional and translational elements. With the PURE system, all elements are known, nuclease and protease activities are reduced, and the concentration of each element can be optimized for maximal protein expression. However, protein expression yield with this system is relatively low due to the consumption of nutrients and energy molecules as well as the accumulation of inhibitory byproducts in the batch format. To enhance protein expression with the PURE system, we developed a feeding solution that was optimized using a miniaturized fluid array device (μFAD) in a continuous-exchange cell-free (CECF) format. The device enabled (1) continuous supply of energy/nutrient molecules from the feeding solution to the reaction solution where protein synthesis occurred, and (2) simultaneous removal of inhibitory expression byproducts from the reaction solution to the feeding solution. Consequently, the synthesis yield of green fluorescent protein (GFP) increased 72.5-fold in comparison with the same reaction in the conventional batch format.

Cell-free protein synthesis (CFPS) enables rapid protein expression for the structural and functional characterization of proteins. Implementation of CFPS in a microfluidic platform has additional benefits such as reduced reaction volumes and simultaneous expression of multiple proteins. Here, we describe a microfluidic device that is composed of 96 continuous-exchange cell-free protein expression units and produces a protein synthesis yield up to 87 times higher than a conventional batch system.

Cell-free protein synthesis (CFPS) enables rapid protein expression for the structural and functional characterization of proteins. Implementation of CFPS in a microfluidic platform has additional benefits such as reduced reaction volumes and simultaneous expression of multiple proteins. Here, we describe a microfluidic device that is composed of 96 continuous-exchange cell-free protein expression units and produces a protein synthesis yield up to 87 times higher than a conventional batch system.

Cell-free protein synthesis (CFPS) has been used as an alternative to cell-based recombinant technology for protein production in academic and industrial labs. The continuous-exchange format generally has higher expression yield by constantly supplying a nutrient solution and removing inhibitory by-products through a porous membrane. Because of the concern of possible membrane clogging by large molecules in the CFPS solution, we investigated the effects of membrane orientation on protein synthesis. We fabricated a miniaturized array device called Vertical-I with its membrane oriented vertically in reference to the table surface and found that the protein synthesis yield in the Vertical-I device was 144% higher than the Horizontal Device reported previously. The reaction time was also faster; β-glucuronidase reached the synthesis yield plateau after 2 h in the Vertical-I device versus 4 h in the Horizontal Device. Possible clogging of membrane pores was confirmed by fluorescein diffusion measurement. Using these results, we designed a device called Vertical-II that would fit into a 96 well plate holder for compatibility with commercial reagent dispensers and microplate readers. The experimentally optimized device increased protein expression 406% over the Horizontal Device and consumes 5 times fewer reagents than a commercial device, showing the potential for high-throughput protein synthesis.

The utility of pathogen and toxin detection systems depends not only upon sensitivity, specificity and capability for multiplexed recognition, but also on access at the point of need, ease of use, and response time. Combining microfluidics and optical biosensors facilitates miniaturization and automation, while careful design makes it possible to test variable quantities of complex matrices such as food (mL) and clinical samples (µL). We report on progress towards an optofluidic system that combines a rotating magnet trap for automated sample processing and a microflow cytometer capable of 4-color analysis to achieve multi-analyte diagnostics. Sample and antibody-coated magnetic beads are introduced into a moving magnetic field created by magnets rotating in the opposite direction of the flow. The color-coded magnetic beads with captured target remain in suspension while fluorescent tracer reagents are introduced to the flow, thus optimizing the binding kinetics and minimizing aggregation. Magnetic beads with bound target and tracer reagents are released for multiplexed analysis by reversing the direction of spinning magnets.

Each Factor 386 36.1.2 How the Forensic Document Examiner Meets Each Factor 39036.2 Conclusion 391 References 391Daubert challenges have become a recurring phenomenon in U.S. courts since the Supreme Court’s seminal decision in 1993, and forensic document examination has been one of the first and most active of the forensic sciences to face such challenges. Prior to the Daubert decision, testimony of the FDE typically consisted of a brief description of professional credentials, followed by an explanation of the examination conducted and conclusions. Post-Daubert testimony requires this and now often a great deal more. FDEs, like all experts, must now be prepared to answer questions regarding the science of their expertise.

PROBLEM: To determine whether cultured human decidual cells and chorion cells produce interleukin-10 (IL-10) after incubation with purified bacterial products. METHOD OF STUDY: Decidual cell cultures and chorion cell cultures were established by standard techniques. With confluence, monolayers of each culture were incubated with purified bacterial products, including various concentrations of lipopolysaccharide (LPS), lipid A, and lipoteichoic acid (LTA) for 16 hr in quadruplicate. Culture supernatants were collected and assayed for immunodetectable IL-10 by enzyme-linked immunoadsorbent assay (ELISA). RESULTS: Both decidual cell cultures and chorion cell cultures produced significant quantities of IL-10 after stimulation with LPS, lipid A, and LTA. Cultures of decidual cells produced more IL-10 than did chorion cell cultures. CONCLUSIONS: Our data indicate that both maternal decidual cells and fetally derived chorion cells can produce IL-10 after incubation with bacterial virulence factors. This finding contrasts with our previous findings in which chorion cells did not produce IL-10 after stimulation with IL-1β, suggesting that chorion cell production after incubation with bacterial products is independent of IL-1β. We speculate that the contribution of anti-inflammatory IL-10 production by human gestational tissues to the inflammatory process in these tissues may be overcome or abrogated by the pro-inflammatory process.