Experimental In-Vivo Models Used in Fat Grafting Research for Volume Augmentation in Soft Tissue Reconstruction

As the popularity of fat grafting research increases, animal models are being used as the source of pre-clinical experimental information for discovery and to enhance techniques. To date, animal models used in this research have not been compared to provide a standardized model. We analyzed publications from 1968-2015 to compare published accounts of animal models in fat grafting research. Data collected included: species used, graft characteristics (donor tissue, recipient area, amount injected, injection technique), time of sacrifice and quantification methods. Mice were most commonly used (56% of studies), with the athymic nude strain utilized most frequently (44%). Autologous fat was the most common source of grafted tissue (52%). Subcutaneous dorsum was the most common recipient site (51%). On average, 0.80+/-0.60 mL of fat was grafted. A single bolus technique was used in 57% of studies. Fat volume assessment was typically completed at the end of the study, occurring at less than 1 week to one year. Graft volume was quantified by weight (63%), usually in conjunction with another analysis. The results demonstrate the current heterogeneity of animal models in this research. We propose that the research community reach a consensus to allow better comparison of techniques and results. One example is the model used in our laboratory and others; this model is described in detail. Eventually, larger animal models may better translate to the human condition but, given increased financial costs and animal facility capability, should be explored when data obtained from small animal studies is exhausted or inconclusive

Generation of Functional Human Adipose Tissue in Mice from Primed Progenitor Cells

Adipose tissue (AT) is used extensively in reconstructive and regenerative therapies, but transplanted fat often undergoes cell death, leading to inflammation, calcification, and requirement for further revision surgery. Previously, we have found that mesenchymal progenitor cells within human AT can proliferate in three-dimensional culture under proangiogenic conditions. These cells (primed ADipose progenitor cells, PADS) robustly differentiate into adipocytes in vitro (ad-PADS). The goal of this study is to determine whether ad-PADS can form structured AT in vivo, with potential for use in surgical applications. Grafts formed from ad-PADS were compared to grafts formed from AT obtained by liposuction after implantation into nude mice. Graft volume was measured by microcomputed tomography scanning, and the functionality of cells within the graft was assessed by quantifying circulating human adiponectin. The degree of graft vascularization by donor or host vessels and the content of human or mouse adipocytes within the graft were measured using species-specific endothelial and adipocyte-specific quantitative real time polymerase chain reaction probes, and histochemistry with mouse and human-specific lectins. Our results show that ad-PADS grafted subcutaneously into nude mice induce robust vascularization from the host, continue to increase in volume over time, express the human adipocyte marker PLIN1 at levels comparable to human AT, and secrete increasing amounts of human adiponectin into the mouse circulation. In contrast, grafts composed of AT fragments obtained by liposuction become less vascularized, develop regions of calcification and decreased content of PLIN1, and secrete lower amounts of adiponectin per unit volume. Enrichment of liposuction tissue with ad-PADS improves vascularization, indicating that ad-PADS may be proangiogenic. Mechanistically, ad-PADS express an extracellular matrix gene signature that includes elements previously associated with small vessel development (COL4A1). Thus, through the formation of a proangiogenic environment, ad-PADS can form functional AT with capacity for long-term survival, and can potentially be used to improve outcomes in reconstructive and regenerative medicine

VARIABILITY STUDY WITH FRIT 510 TO SUPPORT A SECOND TANK 40 DECANT

Sludge Batch 4 (SB4) is currently being processed in the Defense Waste Processing Facility (DWPF) using Frit 510. The slurry pumps in Tank 40 are experiencing in-leakage of bearing water, which is causing the sludge slurry in Tank 40 to become dilute at a rapid rate. Currently, the DWPF is removing this dilution water by performing caustic boiling during the Sludge Receipt and Adjustment Tank (SRAT) cycle. In order to alleviate prolonged SRAT cycle times, which may eventually impact canister production rates, the Liquid Waste Organization (LWO) performed a 100K gallon supernate decant of Tank 40 in April 2008. SRNL performed a supplemental glass variability study to support the April 2008 100K gallon decant incorporating the impact of coupled operations (addition of the Actinide Removal Process (ARP) stream). Recently LWO requested that SRNL assess the impact of a second decant (up to 100K gallon) to the Frit 510-SB4 system. This second decant occurred in June 2008. LWO provided nominal compositions on May 6, 2008 representing Tank 40 prior to the second decant, following the second decant, and the SB4 Heel prior to blending with Tank 51 to constitute SB5. Paper study assessments were performed for these options based on sludge-only and coupled operations processing (ARP addition), as well as possible Na{sub 2}O additions (via NaOH additions) to both flowsheets. A review of the ComProTM database relative to the compositional region defined by the projections after the second decant coupled with Frit 510 identified only a few glasses with similar glass compositions. These glasses were acceptable from a durability perspective, but did not sufficiently cover the new glass compositional region. Therefore, SRNL recommended that a supplemental variability study be performed to support the June 2008 Tank 40 decant. Glasses were selected for the variability study based on three sludge compositional projections (sludge-only, coupled and coupled + 2 wt% Na{sub 2}O) at waste loadings (WLs) of interest to DWPF (32%, 35% and 38%). These nine glasses were fabricated and characterized using chemical composition analysis, X-ray Diffraction (XRD) and the Product Consistency Test (PCT). All of the glasses that were selected for this study satisfy the Product Composition Control System (PCCS) criteria and are deemed processable and acceptable for the DWPF, except for the SB4VS2-03 (sludge-only at 38% WL) target composition. This glass fails the T{sub L} criterion and would not be considered processable based on Slurry Mix Evaporator (SME) acceptability decisions. The durabilities of all of the study glasses (both quenched and ccc) are well below that of the normalized leachate for boron (NL [B]) of the reference EA glass (16.695 g/L) and are predictable using the current PCCS models. Very little variation exists between the NL [B] of the quenched and ccc versions of the glasses. There is some evidence of a trend toward a less durable glass as WL increases for some of the sludge projections. Frit 510 is a viable option for the processing of SB4 after a second Tank 40 decant with or without the addition of products from the ARP stream as well as the 2 wt% Na{sub 2}O addition. The addition of ARP had no negative impacts on the acceptability and predictability of the variability study glasses

INITIAL SLUDGE BATCH 4 TANK 40 DECANT VARIABILITY STUDY WITH FRIT 510

Sludge Batch 4 (SB4) is currently being processed in the Defense Waste Processing Facility (DWPF) using Frit 510. The slurry pumps in Tank 40 are experiencing in-leakage of bearing water, which is causing the sludge slurry feed in Tank 40 to become dilute at a rapid rate. Currently, the DWPF is removing this dilution water by performing caustic boiling during the Sludge Receipt and Adjustment Tank (SRAT) cycle. In order to alleviate prolonged SRAT cycle times that may eventually impact canister production rates, decant scenarios of 100, 150, and 200 kilogallons of supernate were proposed for Tank 40 during the DWPF March outage. Based on the results of the preliminary assessment issued by the Savannah River National Laboratory (SRNL), the Liquid Waste Organization (LWO) issued a Technical Task Request (TTR) for SRNL to (1) perform a more detailed evaluation using updated SB4 compositional information and (2) assess the viability of Frit 510 and determine any potential impacts on the SB4 system. As defined in the TTR, LWO requested that SRNL validate the sludge--only SB4 flowsheet and the coupled operations flowsheet using the 100K gallon decant volume as well as the addition of 3 wt% sodium on a calcined oxide basis. Approximately 12 historical glasses were identified during a search of the ComProTM database that are located within at least one of the five glass regions defined by the proposed SB4 flowsheet options. While these glasses meet the requirements of a variability study there was some concern that the compositional coverage did not adequately bound all cases. Therefore, SRNL recommended that a supplemental experimental variability study be performed to support the various SB4 flowsheet options that may be implemented for future SB4 operations in DWPF. Eighteen glasses were selected based on nominal sludge projections representing the current as well as the proposed flowsheets over a WL interval of interest to DWPF (32-42%). The intent of the experimental portion of the variability study is to demonstrate that the glasses of the Frit 510-modified SB4 compositional region (Cases No.1-5) are both acceptable relative to the Environmental Assessment (EA) reference glass and predictable by the current DWPF process control models for durability. Frit 510 is a viable option for the processing of SB4 after a Tank 40 decant and the addition of products from the Actinide Removal Process (ARP). The addition of ARP did not have any negative impacts on the acceptability and predictability of the variability study glasses. The results of the variability study indicate that all of the study glasses (both quenched and centerline canister cooled (ccc)) have normalized releases for boron that are well below the reference EA glass (16.695 g/L). The durabilities of all of the study glasses are predictable using the current Product Composition Control System (PCCS) durability models with the exception of SB4VAR24ccc (Case No.2 at 41%). PCCS is not applicable to non-homogeneous glasses (i.e. glasses containing crystals such as acmite and nepheline), thus SB4VAR24ccc should not be predictable as it contains nepheline. The presence of nepheline has been confirmed in both SB4VAR13ccc and SB4VAR24ccc by X-ray diffraction (XRD). These two glasses are the first results which indicate that the current nepheline discriminator value of 0.62 is not conservative. The nepheline discriminator was implemented into PCCS for SB4 based on the fact that all of the historical glasses evaluated with nepheline values of 0.62 or greater did not contain nepheline via XRD analysis. Although these two glasses do cause some concern over the use of the 0.62 nepheline value for future DWPF glass systems, the impact to the current SB4 system is of little concern. More specifically, the formation of nepheline was observed in glasses targeting 41 or 42% WL. Current processing of the Frit 510-SB4 system in DWPF has nominally targeted 34% WL. For the SB4 variability study glasses targeting these lower WLs, nepheline formation was not observed and the minimal difference in PCT response between quenched and ccc versions supported its absence

Microbial transformations of selenite by methane-oxidizing bacteria

Abstract Methane oxidizing bacteria are well known for their role in the global methane cycle and their potential for microbial transformation of wide range of hydrocarbon and chlorinated hydrocarbon pollution. Recently, it has also emerged that methane-oxidizing bacteria interact with inorganic pollutants in the environment. Here we report what we believe to be the first study of the interaction of pure strains of methane-oxidizing bacteria with selenite. Results indicate that the commonly used laboratory model strains of methane oxidizing bacteria, Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b are both able to reduce the toxic selenite (SeO32-) but not selenate (SeO42-) to red spherical nanoparticulate elemental selenium (Se0), which was characterised via EDX and EXAFS. The cultures also produced volatile selenium-containing species, which suggests that both strains may have an additional activity that can either transform Se0 or selenite into volatile methylated forms of selenium. Transmission electron microscopy (TEM) measurements and experiments with the cell fractions: cytoplasm, cell wall and cell membrane show that the nanoparticles are formed mainly on the cell wall. Collectively these results are promising for the use of methane-oxidizing bacteria for bioremediation or suggest possible uses in the production of selenium nanoparticles for biotechnology

Evidence-based Kernels: Fundamental Units of Behavioral Influence

This paper describes evidence-based kernels, fundamental units of behavioral influence that appear to underlie effective prevention and treatment for children, adults, and families. A kernel is a behavior–influence procedure shown through experimental analysis to affect a specific behavior and that is indivisible in the sense that removing any of its components would render it inert. Existing evidence shows that a variety of kernels can influence behavior in context, and some evidence suggests that frequent use or sufficient use of some kernels may produce longer lasting behavioral shifts. The analysis of kernels could contribute to an empirically based theory of behavioral influence, augment existing prevention or treatment efforts, facilitate the dissemination of effective prevention and treatment practices, clarify the active ingredients in existing interventions, and contribute to efficiently developing interventions that are more effective. Kernels involve one or more of the following mechanisms of behavior influence: reinforcement, altering antecedents, changing verbal relational responding, or changing physiological states directly. The paper describes 52 of these kernels, and details practical, theoretical, and research implications, including calling for a national database of kernels that influence human behavior