Bone-Conditioned Medium Changes Gene Expression in Bone-Derived Fibroblasts.

PURPOSE Autologous bone is used for augmentation in the course of oral implant placement. Bone grafts release paracrine signals that can modulate mesenchymal cell differentiation in vitro. The detailed genetic response of the bone-derived fibroblasts to these paracrine signals has remained elusive. Paracrine signals accumulate in bone-conditioned medium (BCM) prepared from porcine cortical bone chips. MATERIALS AND METHODS In this study, bone-derived fibroblasts were exposed to BCM followed by a whole genome expression profiling and downstream quantitative reverse transciptase polymerase chain reaction of the most strongly regulated genes. RESULTS The data show that ADM, IL11, IL33, NOX4, PRG4, and PTX3 were differentially expressed in response to BCM in bone-derived fibroblasts. The transforming growth factor beta (TGF-β) receptor 1 antagonist SB431542 blocked the effect of BCM on the expression of the gene panel, except for IL33. CONCLUSION These in vitro results extend existing evidence that cortical bone chips release paracrine signals that provoke a robust genetic response in mesenchymal cells that is not exclusively mediated via the TGF-β receptor. The present data provide further insights into the process of graft consolidation.

Conditioned medium from fresh and demineralized bone enhances osteoclastogenesis in murine bone marrow cultures

OBJECTIVES Osteoclasts rapidly form on the surface of bone chips at augmentation sites. The underlying molecular mechanism, however, is unclear. Soluble factors released from bone chips in vitro have a robust impact on mesenchymal cell differentiation. Whether these soluble factors change the differentiation of hematopoietic cells into osteoclasts remains unknown. METHODS Osteoclastogenesis, the formation of tartrate-resistant acid phosphatase-positive multinucleated cells, was studied with murine bone marrow cultures exposed to RANKL and M-CSF, and conditioned medium from fresh (BCM) and demineralized bone matrix (DCM). Histochemical staining, gene and protein expression, as well as viability assays were performed. RESULTS This study shows that BCM had no impact on osteoclastogenesis. However, when BCM was heated to 85°C (BCMh), the number of tartrate-resistant acid phosphatase-positive multinucleated cells that developed in the presence of RANKL and M-CSF approximately doubled. In line with the histochemical observations, there was a trend that BCMh increased expression of osteoclast marker genes, in particular the transcription factor c-fos. The expression of c-fos was significantly reduced by the TGF-β receptor I antagonist SB431542. DCM significantly stimulated osteoclastogenesis, independent of thermal processing. CONCLUSIONS These data demonstrate that activated BCM by heat and DBM are able to stimulate osteoclastogenesis in vitro. These in vitro results support the notion that the resorption of autografts may be supported by as yet less defined paracrine mechanisms.

Effects of Antiseptic Solutions Commonly Used in Dentistry on Bone Viability, Bone Morphology, and Release of Growth Factors

PURPOSE Antiseptic solutions are commonly used in dentistry for a number of sterilization procedures, including harvesting of bone chips, irrigation of extraction sockets, and sterilization of osteonecrotic bone. Despite its widespread use, little information is available regarding the effects of various antiseptic solutions on bone cell viability, morphology, and the release of growth factors. MATERIALS AND METHODS The antiseptic solutions included 1) 0.5% povidone iodine (PI), 2) 0.2% chlorhexidine diguluconate (CHX), 3) 1% hydrogen peroxide (H2O2), and 4) 0.25% sodium hypochlorite (HYP). Bone samples collected from porcine mandibular cortical bone were rinsed in the antiseptic solutions for 10 minutes and assessed for cell viability using an MTS assay and protein release of transforming growth factor (TGF-β1), bone morphogenetic protein 2 (BMP2), vascular endothelial growth factor (VEGF), interleukin (IL)-1β, and receptor activator of nuclear factor κB ligand (RANKL) using an enzyme-linked immunosorbent assay at 15 minutes and 4 hours after rinsing. RESULTS After antiseptic rinsing, changes to the surface protein content showed marked alterations, with an abundant protein layer remaining on CHX-rinsed bone samples. The amount of surface protein content gradually decreased in the following order: CHX, H2O2, PI, and HYP. A similar trend was also observed for the relative cell viability from within bone samples after rinsing, with up to 6 times more viable cells found in the CHX-rinsed bone samples than in the HYP- and PI-rinsed samples. An analysis of the growth factors found that both HYP and PI had significantly lower VEGF and TGF-β1 protein release from bone samples at 15 minutes and 4 hours after rinsing compared with CHX and H2O2. A similar trend was observed for RANKL and IL-1β protein release, although no change was observed for BMP2. CONCLUSIONS The results from the present study have demonstrated that antiseptic solutions present with very different effects on bone samples after 10 minutes of rinsing. Rinsing with CHX maintained significantly higher cell viability and protein release of growth factors potent to the bone remodeling cycle.

The art of dieting: Exposure to thin sculptures effortlessly reduces the intake of unhealthy food in motivated eaters

Thin, human-like sculptures by the artist Alberto Giacometti, applied as environmental cues, have been found to facilitate dieting by reducing chocolate intake and promoting healthy snack choices. However, the processes underlying this “Giacometti effect” have been left unexplored so far. The present study therefore first examines the effortlessness of the effect. More specifically, it aims to determine whether the sculptures reduce unhealthy food intake when only few cognitive resources for their influence are available. For this purpose, the participants in a chip tasting were given the cognitive load task of memorizing either 10 or two digits during the tasting. The results indicate that the sculptures reduced participants’ chip intake independent of the cognitive load. Thus, they influenced participants’ eating behavior even when only few cognitive resources were available. The results also indicate that the sculptures reduced chip intake only when the participants liked the chips. The sculptures could thus exert their influence when individuals were motivated to eat and the dieting cues were useful. The finding that the Giacometti sculptures, applied as environmental dieting or health cues, influenced individuals when only few cognitive resources were available, could indicate a crucial advantage for the application of these cues in complex, real-world settings.

Spatio-temporal co-ordination of RhoA, Rac1 and Cdc42 activation during prototypical edge protrusion and retraction dynamics

The three canonical Rho GTPases RhoA, Rac1 and Cdc42 co-ordinate cytoskeletal dynamics. Recent studies indicate that all three Rho GTPases are activated at the leading edge of motile fibroblasts, where their activity fluctuates at subminute time and micrometer length scales. Here, we use a microfluidic chip to acutely manipulate fibroblast edge dynamics by applying pulses of platelet-derived growth factor (PDGF) or the Rho kinase inhibitor Y-27632 (which lowers contractility). This induces acute and robust membrane protrusion and retraction events, that exhibit stereotyped cytoskeletal dynamics, allowing us to fairly compare specific morphodynamic states across experiments. Using a novel Cdc42, as well as previously described, second generation RhoA and Rac1 biosensors, we observe distinct spatio-temporal signaling programs that involve all three Rho GTPases, during protrusion/retraction edge dynamics. Our results suggest that Rac1, Cdc42 and RhoA regulate different cytoskeletal and adhesion processes to fine tune the highly plastic edge protrusion/retraction dynamics that power cell motility.

Impacts of Technology Adoption: Comparing Returns to the Farming Sector in Maine under Alternative Technology Regimes

This thesis tests if certain technology choices are associated with a reduction in the proportion of farming activities in the agro-food system in Maine. Goodman, Sorj, and Wilkinson define appropriationism as the replacement of farming sector activities by industrial inputs. Based on the concept of appropriationism, industrial fanning systems using large amounts of synthetic inputs contribute less to fanning than more agrarian systems, like organic fanning. Thus, returns to the farming sector should be greater for organic compared with conventional potato fanning in Maine since organic farming uses fewer industrial inputs. Goodman et. al. define substitutionism as the displacement of farming sector commodities and activities by industrial processes in the marketing sector. Based on the concept of substitutionism, returns to the farming sector should be greater for Lay's Classic®™ potato chips made from natural potatoes compared with Baked Lay's®™ potato crisps manufactured from processed dehydrated potatoes. Returns to the farming sector are defined as returns to the farmer or farm family from farming activities, returns to farm labor, and returns to farmers and farm labor producing inputs used on the farm. Results show absolute returns to the farming sector are less for organic compared to conventional tablestock potato farms in Maine. However as a proportion of farm revenues, large organic farms that market at least 25% of their produce to retail stores or directly to consumers do as well as conventional farms. When comparing returns as a proportion of consumer expenditures, these organic farms do better than conventional farms. Returns to the farming sector are less for organic because of yield penalties, cost of marketing services, and diseconomies of size for organic tablestock potato farms. Expanding acreage and reintegrating livestock with cropping systems may increase returns to the fanning sector. Organic farming demonstrates difficulties in providing marketing services at the farm level. Providing marketing services limits the ability to expand production to capture economies of size. Maine organic potato farmers emphasize non-monetary values such as supporting sustainable agriculture, self-sufficiency, the intrinsic value of work, and close community and family connections. Returns to the farming sector as a proportion of consumer expenditures are about three times greater for Lay's Classic®™ potato chips than for Baked Lay's®™ potato crisps, since the value that farmers receive for potatoes used to produce dehydrated potato flakes in one pound of crisps is about half of the value that farmers receive for potatoes used to make one pound of chips. However, this assumes farmers assign a cost to producing low-grade potatoes for dehydration proportionate to their value. Premium potatoes are used to produce potato chips. Low-grade potatoes are used to produce the dehydrated potato flakes used to make potato crisps. Returns to the farming sector are slightly greater for potato crisps if no costs are allocated to producing low-grade potatoes for dehydration. A shift in consumer preferences from potato chips to crisps may result in a geographical shift of potato production from Maine to the Pacific Northwest assuming no food-grade dehydration facilities are built in Maine.

Dielectrophoresis-based analyte separation and analysis

Dielectrophoresis—the tendency of a material of high dielectric permittivity to migrate in an electrical field gradient to a region of maximum field strength—provides an ideal motive force for manipulating small volumes of biological analytes in microfluidic microsystems. The work described in this thesis was based on the hypothesis that dielectrophoresis could be exploited to provide high-resolution cell separations in microsystems as well as a means for the electrically-controllable manipulation of solid supports for molecular analysis. To this end, a dielectrophoretic/gravitational field-flow-fractionation (DEP/G-FFF) system was developed and the separation performance evaluated using various types and sizes of polystyrene microspheres as model particles. It was shown that separation of the polystyrene beads was based on the differences in their effective dielectrophoretic properties. The ability of an improved DEP/G-FFF system to separate genetically identical, but phenotypically dissimilar cell types was demonstrated using mixtures of 6m2 mutant rat kidney cells grown under transforming and non-transforming culture conditions. Additionally, a panel of engineered dielectric microspheres was designed with specific, predetermined dielectrophoretic properties such that their dielectrophoretic behaviors would be controllable and predictable. The fabrication method involved the use of gold-coated polystyrene microsphere cores coated with a self-assembled monolayer of alkanethiol and, optionally, a self-assembled monolayer of phospholipid to form a thin-insulating-shell-over-conductive-interior structure. The successful development of the DEP/G-FFF separation system and the dielectrically engineered microspheres provides proof-of-principle demonstrations of enabling dielectrophoresis-based microsystem technology that should provide powerful new methods for the manipulation, separation and identification of analytes in many diverse fields. ^

Los Voluntarios Abren les Puertas: las Cinco Claves para el Manejo Seguro del Plomo

Millions of houses and apartments built before 1978 have paint that contains lead. Chips, dust, and fumes from this paint can be very dangerous if they are not handled properly. Lead is particularly hazardous to unborn babies, infants, and young children. Volunteers in painting and housing-rehabilitation programs often work in homes that contain lead paint. The work they perform can create a lead hazard if they disturb this paint and produce paint chips or dust. Volunteers Opening Doors is a video program for these volunteers. It explains how volunteers can protect housing residents,themselves, and their families from lead poisoning by using the five keys to lead safety: 1. Protect residents and their belongings. 2. Prepare the work area. 3. Protect yourself from dust and debris. 4. Work wet. 5. Work clean.

Volunteers Opening Doors: the Five Keys to Lead Safety

Millions of houses and apartments built before 1978 have paint that contains lead. Chips, dust, and fumes from this paint can be very dangerous if they are not handled properly. Lead is particularly hazardous to unborn babies, infants, and young children. Volunteers in painting and housing-rehabilitation programs often work in homes that contain lead paint. The work they perform can create a lead hazard if they disturb this paint and produce paint chips or dust. Volunteers Opening Doors is a video program for these volunteers. It explains how volunteers can protect housing residents,themselves, and their families from lead poisoning by using the five keys to lead safety: 1. Protect residents and their belongings. 2. Prepare the work area. 3. Protect yourself from dust and debris. 4. Work wet. 5. Work clean.

An investigation into the relationship between screen time, consumption of advertised foods, and physical activity among Texas 4th grade elementary school children

Background. The high prevalence of obesity among children has spurred creation of a list of possible causative factors, including the advertising of foods of minimal nutritional value, a decrease in physical activity, and increased media use. Few studies show prevalence rates of these factors among large cohorts of children. ^ Methods. Using data from the 2004-2005 School Physical Activity and Nutrition project (SPAN), a secondary analysis of 7907 4th-grade children (mean age 9.74 years) was conducted. In addition, a comic-book–based intervention that addressed advertised food consumption, physical activity, and media use was developed and evaluated using a pre-post test design among 4th-grade children in an urban school district. ^ Results. Among a cohort of 4th-grade children across the state of Texas, children who had more than 2 hours of video game or computer time the previous day were more than twice as likely to drink soda and eat candy or pastries. In addition, children who watched more than 2 hours of TV the previous day were more than three times as likely to consume chips, punch, soda, candy, frozen desserts, or pastries (AOR 3.41, 95% CI: 1.58, 7.37). A comic-book based intervention held great promise and acceptance among 4th-grade children. Outcome evaluation showed that while results moved in a positive direction, they were not statistically significant. ^ Conclusion. Statistically significant associations were found between screen time and eating various types of advertised food. The comic book intervention was widely accepted by the children exposed to it, and pre-post surveys indicated they moved constructs in a positive direction. Further research is needed to look at more specific ways in which children are exposed to TV, and the relationship of the TV viewing time with their consumption of advertised foods. In addition, researchers should look at comic book interventions more closely and attempt to utilize them in more in studies with a longer follow-up time. ^

Expression study of the atherosclerosis mouse aorta reveals significant disturbance of calcium signaling pathway

Atherosclerosis is a complex disease resulting from interactions of genetic and environmental risk factors leading to heart failure and stroke. Using an atherosclerotic mouse model (ldlr-/-, apobec1-/- designated as LDb), we performed microarray analysis to identify candidate genes and pathways, which are most perturbed in changes in the following risk factors: genetics (control C57BL/6 vs. LDb mice), shearstress (lesion-prone vs. lesion-resistant regions in LDb mice), diet (chow vs. high fat fed LDb mice) and age (2-month-old vs. 8-month old LDb mice). ^ Atherosclerotic lesion quantification and lipid profile studies were performed to assess the disease phenotype. A microarray study was performed on lesion-prone and lesion-resistant regions of each aorta. Briefly, 32 male C57BL/6 and LDb mice (n =16/each) were fed on either chow or high fat diet, sacrificed at 2- and 8-months old, and RNA isolated from the aortic lesion-prone and aortic lesion-resistant segments. Using 64 Affymetrix Murine 430 2.0 chips, we profiled differentially expressed genes with the cut off value of FDR ≤ 0.15 for t-test, and q <0.0001 for the ANOVA. The data were normalized using two normalization methods---invariant probe sets (Loess) and Quantile normalization, the statistical analysis was performed using t-tests and ANOVA, and pathway characterization was done using Pathway Express (Wayne State). The result identified the calcium signaling pathway as the most significant overrepresented pathway, followed by focal adhesion. In the calcium signaling pathway, 56 genes were found to be significantly differentially expressed out of 180 genes listed in the KEGG calcium signaling pathway. Nineteen of these genes were consistently identified by both statistical tests, 11 of which were unique to the test, and 26 were unique to the ANOVA test, using the cutoffs noted above. ^ In conclusion, this finding suggested that hypercholesterolemia drives the disease progression by altering the expression of calcium channels and regulators which subsequently results in cell differentiation, growth, adhesion, cytoskeletal change and death. Clinically, this pathway may serve as an important target for future therapeutic intervention, and thus the calcium signaling pathway may serve as an important target for future diagnostic and therapeutic intervention. ^

Assessing how middle school students' nutrient intake varies based on the availability of a la carte offerings at lunch

Purpose: School districts in the U.S. regularly offer foods that compete with the USDA reimbursable meal, known as `a la carte' foods. These foods must adhere to state nutritional regulations; however, the implementation of these regulations often differs across districts. The purpose of this study is to compare two methods of offering a la carte foods on student's lunch intake: 1) an extensive a la carte program in which schools have a separate area for a la carte food sales, that includes non-reimbursable entrees; and 2) a moderate a la carte program, which offers the sale of a la carte foods on the same serving line with reimbursable meals. ^ Methods: Direct observation was used to assess children's lunch consumption in six schools, across two districts in Central Texas (n=373 observations). Schools were matched on socioeconomic status. Data collectors were randomly assigned to students, and recorded foods obtained, foods consumed, source of food, gender, grade, and ethnicity. Observations were entered into a nutrient database program, FIAS Millennium Edition, to obtain nutritional information. Differences in energy and nutrient intake across lunch sources and districts were assessed using ANOVA and independent t-tests. A linear regression model was applied to control for potential confounders. ^ Results: Students at schools with extensive a la carte programs consumed significantly more calories, carbohydrates, total fat, saturated fat, calcium, and sodium compared to students in schools with moderate a la carte offerings (p<.05). Students in the extensive a la carte program consumed approximately 94 calories more than students in the moderate a la carte program. There was no significant difference in the energy consumption in students who consumed any amount of a la carte compared to students who consumed none. In both districts, students who consumed a la carte offerings were more likely to consume sugar-sweetened beverages, sweets, chips, and pizza compared to students who consumed no a la carte foods. ^ Conclusion: The amount, type and method of offering a la carte foods can significantly affect student dietary intake. This pilot study indicates that when a la carte foods are more available, students consume more calories. Findings underscore the need for further investigation on how availability of a la carte foods affects children's diets. Guidelines for school a la carte offerings should be maximized to encourage the consumption of healthful foods and appropriate energy intake.^

Student consumption of a la carte foods during school lunch meals

The purpose of this study was to evaluate students' lunch consumption compared to NSLP guidelines, the contribution of competitive foods to calorie intake at lunch, and the differences in nutrient and food group intake between the a la carte food consumers and non- a la carte food consumers.^ In Fall 2011, 1170 elementary and 440 intermediate students were observed anonymously during school lunch. The foods eaten, their source, grade level, and gender were recorded. All a la carte offerings met the Texas School Nutrition Policy.^ Differences in nutrient and food group intake by grade level and between students who consumed a la carte and those who did not were assessed using ANCOVA. A chi-squared analysis was conducted to evaluate differences in a la carte food consumption by grade level, gender, and the school's low income status.^ Average lunch intakes for elementary students were 457 (SD 164) calories for elementary students and 541 calories (SD 188) for intermediate students (p<0.001). 760 students (47%) consumed 937 a la carte foods, with the most often consumed items being chips (32%), ice cream (22%) and snack items (18%). Mean a la carte food intakes were 60 and 98 calories for elementary and intermediate schools respectively (p<0.001). Significantly more (p<0.000) intermediate students (34.3%) consumed a la carte items compared to elementary students (27.5%).^ Students who consumed a la carte foods had significantly higher intakes of calories (p<0.000), fat (p<0.000), sodium (p<0.002), fiber (p<0.000), added sugar (p<0.000), total grains (p<0.000), dessert foods (p<0.000), and snack chips (p<0.000) and lower intakes of vitamin A (p<0.001), iron (p<0.000), fruit (p<0.022), vegetables (p<0.031), milk (p<0.000), and juice (p<0.000) compared to students who did not eat a la carte foods.^ Although previous studies have found that reducing availability of unhealthy items at school decreased student consumption of these items, the results of this study indicate that even the strict guidelines set forth by the state of Texas are not sufficient to prevent increased caloric intake and poor nutrient intake. Strategies to improve student selection and consumption at school lunch when a la carte foods are available are warranted.^

Dietary patterns and body mass index of Canadian Inuit communities in 1999 and 2008

Objectives: The International Polar Year (IPY) Inuit Health Survey provided an opportunity to compare dietary and body mass index (BMI) data with data collected a decade earlier for the same communities. Study design: A dietary survey included 1,929 randomly selected participants aged 15 years or older, selected from 18 Inuit communities in 1998-1999. The IPY survey included 2,595 randomly selected participants aged 18 years or older, selected from 36 Inuit communities in 2007-2008. Data from the same 18 communities included in both surveys were compared for adults 20 years and older. Methods: Twenty-four-hour dietary recall data were analysed to assess the percentage of energy from traditional and market foods by sex and age groups. Body mass index (BMI) was assessed to establish the prevalence of obesity by sex and age groups in both surveys. Results: There was a significant decrease (p<=0.05) in energy contribution from traditional food and a significant increase in market food consumption over time. Sugar-sweetened beverages, chips and pasta all increased as percentages of energy. BMI increased overall for women and for each age stratum evaluated (p<0.05). Conclusion: The nutrition transition continues in the Canadian Arctic with a concurrent increase in BMI.

Pebbles and carbonate nodules Legs 56-57 Holes

At all DSDP Leg 56 drilling sites, exotic pebbles occur commonly, throughout the cores. Chips of carbonate nodules occur only at Site 434 on the lower inner trench wall. Both exotic pebbles and carbonate nodule chips sometimes tend to be concentrated at particular levels of cores. Exotic pebbles are generally well rounded and consist of various rock types, such as dacite, andesite, basalt, tuff, gabbro, granodiorite, metaquartzite, biotite hornfels, lithic wacke, mudstone, etc., of which dacite occurs commonly at all the sites. Almost all pebbles at Site 436 and most at Sites 434 and 435 may have been rafted by ice. Some at the latter sites may have been derived by down-slope slumping. Carbonate nodules consist of microcrystalline dolomite, manganoan calcite, and siderite; CaCO3 content ranges from 22 to 65 per cent. They are also generally characterized by a high content of P2O5. The nodules are commonly rich in diatom remains, some of which indicate that the nodules are autochthonous. Some nodules contain abundant glass shards, with a modal refractive index of 1.499, almost identical to shards in the surrounding mud and ooze. These facts suggest that the carbonate nodules may have been formed diagenetically, in situ. This may throw light on problems of the formation of carbonate nodules in ancient "geosynclinal" sediments. It is also very important to point out that these carbonate nodules were formed within sediment deposited well below the CCD.