ObjectiveThe study of obesity-related metabolic syndrome or Type 2 diabetes (T2D) in children is particularly difficult because of fear of needles. We tested a non-invasive approach to study inflammatory parameters in an at-risk population of children to provide proof-of-principle for future investigations of vulnerable subjects.Design and MethodsWe evaluated metabolic differences in 744, 11-year old children selected from underweight, normal healthy weight, overweight and obese categories by analyzing fasting saliva samples for 20 biomarkers. Saliva supernatants were obtained following centrifugation and used for analyses.ResultsSalivary C-reactive protein (CRP) was 6 times higher, salivary insulin and leptin were 3 times higher, and adiponectin was 30% lower in obese children compared to healthy normal weight children (all P<0.0001). Categorical analysis suggested that there might be three types of obesity in children. Distinctly inflammatory characteristics appeared in 76% of obese children while in 13%, salivary insulin was high but not associated with inflammatory mediators. The remaining 11% of obese children had high insulin and reduced adiponectin. Forty percent of the non-obese children were found in groups which, based on biomarker characteristics, may be at risk for becoming obese.ConclusionsSignificantly altered levels of salivary biomarkers in obese children from a high-risk population, suggest the potential for developing non-invasive screening procedures to identify T2D-vulnerable individuals and a means to test preventative strategies.
Evidence-based practice is the procedure whereby clinicians incorporate best research evidence, clinical expertise, and patient values to provide best patient care. Recently, there has been a significant push towards occupational therapists' adoption of evidence-based practice. This systematic review aimed to determine occupational therapists' attitudes, knowledge, and utilization of evidence-based practice. Method: A search of literature published between 2000-12 was conducted in relation to occupational therapists' practice. Academic Search Complete, Cumulative Index of Nursing and Allied Health Literature Plus, PsycARTICLES, Ingenta, Medline, Science Direct, and Journal Storage were systematically searched using MeSH and free-text keywords. Google Scholar and reference lists were also searched. Findings: Thirty-two papers were selected for review: 23 were quantitative, 8 were qualitative, and 1 used a mixed methods design. Studies demonstrated that occupational therapists hold positive attitudes towards evidence-based practice. However, these attitudes do not translate into practice, with research indicating a lack of evidence-based practice utilization. Occupational therapists perceive a number of barriers to evidence-based practice, including lack of time, lack of availability and accessibility of research, and having limited research skills. Conclusion: It is essential that educational and training initiatives provide therapists with the tools and support they need to engage fully with research evidence and its application within clinical care.
Multiplexing arrays increase the throughput and decrease sample requirements for studies employing multiple biomarkers. The goal of this project was to examine the performance of Multiplex arrays for measuring multiple protein biomarkers in saliva and serum. Specimens from the OsteoPerio ancillary study of the Women’s Health Initiative Observational Study were used. Participants required the presence of at least 6 teeth and were excluded based on active cancer and certain bone issues but were not selected on any specific condition. Quality control (QC) samples were created from pooled serum and saliva. Twenty protein markers were measured on five multiplexing array panels. Sample pretreatment conditions were optimized for each panel. Recovery, lower limit of quantification (LLOQ) and imprecision were determined for each analyte. Statistical adjustment at the plate level was used to reduce imprecision estimates and increase the number of usable observations. Sample pre-treatment improved recovery estimates for many analytes. The LLOQ for each analyte agreed with manufacturer specifications except for MMP-1 and MMP-2 which were significantly higher than reported. Following batch adjustment, 17 of 20 biomarkers in serum and 9 of 20 biomarkers in saliva demonstrated acceptable precision, defined as <20% coefficient of variation (<25% at LLOQ). The percentage of cohort samples having levels within the reportable range for each analyte varied from 10% to 100%. The ratio of levels in saliva to serum varied from 1∶100 to 28∶1. Correlations between saliva and serum were of moderate positive magnitude and significant for CRP, MMP-2, insulin, adiponectin, GM-CSF and IL-5. Multiplex arrays exhibit high levels of analytical imprecision, particularly at the batch level. Careful sample pre-treatment can enhance recovery and reduce imprecision. Following statistical adjustments to reduce batch effects, we identified biomarkers that are of acceptable quality in serum and to a lesser degree in saliva using Multiplex arrays.
1. In order to exclude the possibility of differences in maternal care which are known to result from typical methods of undernutrition during the suckling period, rat pups were reared artificially on different planes of nutrition away from their mothers.2. Artificial rearing was accomplished by fitting infant rats with a gastric cannula through which a milk substitute was infused intermittently. Rats were fed thus from 4 to 21 d on a high (ARHI) or a low (ARLO; 44%of ARHI level) plane of nutrition. Underfeeding of the ARLO group was continued till 25 d, after which all rats were given a good-quality pelleted diet nd lib. 3. Compared with mother-reared (MR) litter-mates, ARHI rats showed advanced eye-opening and, at 21 and 25 d, they resisted restraint more strongly.4. Growth in body-weight of ARHI and MR rats was similar but, when autopsied at 32 weeks, the ARHI rats were shorter (nose-rump length) and had lighter gastrocnemius muscles, adrenals and brains, but heavier epididymal-fat pads.5. ARLO rats had deficits at 32 weeks compared with ARHI rats in whole body, kidney and epididymal-fat-pad weights, and in tibia length.6. In a second experiment, ARHI and MR rats were killed at 21 d. All the differences found at 32 weeks were already present at 21 d. In addition, the ARHI pups had enlarged livers and intestines but shorter tibias.7. The milk substitute, which is one commonly used in such studies, has a low protein and high carbohydrate content compared with rats' milk. This difference probably caused the abnormal organ growth of ARHI rats.
Periodontal disease (PD) has been suggested to be a risk factor for Alzheimer’s disease (AD). We tested the impact of ligature-induced PD on 5xFAD mice and WT littermates. At baseline, 5xFAD mice presented significant alveolar bone loss compared to WT mice. After the induction of PD, both WT and 5xFAD mice experienced alveolar bone loss. PD increased the level of Iba1-immunostained microglia in WT mice. In 5xFAD mice, PD increased the level of insoluble Aβ42. The increased level in Iba1 immunostaining that parallels the accumulation of Aβ in 5xFAD mice was not affected by PD except for a decrease in the dentate gyrus. Analysis of double-label fluorescent images showed a decline in Iba1 in the proximity of Aβ plaques in 5xFAD mice with PD compared to those without PD suggesting a PD-induced decrease in plaque-associated microglia (PAM). PD reduced IL-6, MCP-1, GM-CSF, and IFN-γ in brains of WT mice and reduced IL-10 in 5xFAD mice. The data demonstrated that PD increases neuroinflammation in WT mice and disrupts the neuroinflammatory response in 5xFAD mice and suggest that microglia is central to the association between PD and AD.
These findings suggest that different strains of F. nucleatum impact neutrophil function in different ways. Two of three subspecies blocked neutrophil superoxide generation in response to a secondary stimulus, preventing oxidative killing by neutrophils. The direct role of bridging species in pathogenesis of periodontitis may be greater than previously suspected in which they create a favorable environment for pathogenic transition of the dental ecosystem.
Lysyl oxidase propeptide (LOX-PP) ectopic overexpression inhibits the growth of cancer xenografts. Here the ability and mode of action of purified recombinant LOX-PP (rLOX-PP) protein to inhibit the growth of pre-existing xenografts was determined. Experimental approaches employed were direct intratumoral injection (i.t.) of rLOX-PP protein into murine breast cancer NF639 xenografts, and application of a slow release formulation of rLOX-PP implanted adjacent to tumors in NCR nu/nu mice (n = 10). Tumors were monitored for growth, and after sacrifice were subjected to immunohistochemical and Western blot analyses for several markers of proliferation, apoptosis, and for rLOX-PP itself. Direct i.t. injection of rLOX-PP significantly reduced tumor volume on days 20, 22 and 25 and tumor weight at harvest on day 25 by 30% compared to control. Implantation of beads preloaded with 35 micrograms rLOX-PP (n = 10) in vivo reduced tumor volume and weight at sacrifice when compared to empty beads (p<0.05). A 30% reduction of tumor volume on days 22 and 25 (p<0.05) and final tumor weight on day 25 (p<0.05) were observed with a reduced tumor growth rate of 60% after implantation. rLOX-PP significantly reduced the expression of proliferation markers and Erk1/2 MAP kinase activation, while prominent increases in apoptosis markers were observed. rLOX-PP was detected by immunohistochemistry in harvested rLOX-PP tumors, but not in controls. Data provide pre-clinical findings that support proof of principle for the therapeutic anti-cancer potential of rLOX-PP protein formulations.
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