Quantifying microbial utilization of petroleum hydrocarbons in salt-marsh sediments using the ^(13)C content of bacterial rRNA

Natural remediation of oil spills is catalyzed by complex microbial consortia. Here we take a whole-community approach to investigate bacterial incorporation of petroleum hydrocarbons from a simulated oil spill. We utilized the natural difference in carbon-isotopic abundance between a salt marsh ecosystem supported by the ^(13)C-enriched C4 grass, Spartina alterniflora, and the ^(13)C-depleted composition of petroleum to monitor changes in the ^(13)C content of biomass. Magnetic-bead capture methods for the selective recovery of bacterial RNA were used to monitor the ^(13)C content of bacterial biomass during a two-week experiment. The data show that by the end of the experiment, up to 26% of bacterial biomass derived from consumption of the freshly-spilled oil. The results contrast with the inertness of a nearby relict spill, which occurred in 1969 in West Falmouth, MA. Sequences of 16S rRNA genes from our experimental samples also were consistent with previous reports suggesting the importance of {gamma}- and {delta}-Proteobacteria and Firmicutes in the remineralization of hydrocarbons. The magnetic-bead capture approach makes it possible to quantify uptake of petroleum hydrocarbons by microbes in-situ. Although employed here at the Domain level, RNA-capture procedures can be highly specific. The same strategy could be used with genus-level specificity, something which is not currently possible using the ^(13)C content of biomarker lipids

Preventing Teen Relationship Abuse and Sexual Assault through Bystander Training: Intervention Outcomes for School Personnel

The purpose of the current study was to examine the impact of exposure to Bringing in the Bystander—High School Curriculum (BITB-HSC) on school personnel, which included a seven session classroom curriculum for ninth through twelfth graders (student curriculum), a bystander training workshop for school personnel (school personnel workshop), and reading materials (handout). We examined how exposure to these various BITB-HSC intervention components was associated with school personnel’s knowledge and bystander efficacy, intentions, and barriers specific to student relationship abuse (RA) and sexual assault (SA). Participants were 488 school personnel from 12 high schools in upper New England who completed the 4-month follow-up survey that assessed for intervention exposure (284 participants completed both the baseline and follow-up survey). Whereas 53% of participants were exposed to no intervention components, the other half of the sample were exposed to a combination of intervention components. Higher baseline knowledge and reactive bystander intentions were associated with subsequent exposure to both the student curriculum and the handout, and fewer barriers to bystander action predicted exposure to the school personnel workshop. Exposure to the school personnel workshop, student curriculum, and handout was associated with subsequent greater knowledge, exposure to the student curriculum predicted reactive bystander intentions, and exposure to the handout predicted higher reactive bystander intentions and bystander efficacy. Findings suggest that despite challenges with engagement, exposure to the BITB-HSC components may be a useful tool in improving school personnel’s responses to RA and SA among high school students

Enhanced Recovery after Surgery in a Single High-Volume Surgical Oncology Unit: Details Matter

Benefits of ERAS protocol have been well documented; however, it is unclear whether the improvement stems from the protocol or shifts in expectations. Interdisciplinary educational seminars were conducted for all health professionals. However, one test surgeon adopted the protocol. 394 patients undergoing elective abdominal surgery from June 2013 to April 2015 with a median age of 63 years were included. The implementation of ERAS protocol resulted in a decrease in the length of stay (LOS) and mortality, whereas the difference in cost was found to be insignificant. For the test surgeon, ERAS was associated with decreased LOS, cost, and mortality. For the control providers, the LOS, cost, mortality, readmission rates, and complications remained similar both before and after the implementation of ERAS. An ERAS protocol on the single high-volume surgical unit decreased the cost, LOS, and mortality

Shallow-water benthic habitats of southwest Puerto Rico

This report describes the creation and assessment of benthic habitat maps for shallow-water (<30m) marine environments of the Guánica/Parguera and Finca Belvedere Natural Reserve in southwest Puerto Rico. The objective was to provide spatially-explicit information on the habitat types, biological cover and live coral cover of the region’s coral reef ecosystem. These fine-scale habitat maps, generated by interpretation of 2010 satellite imagery, provide an update to NOAA’s previous digital maps of the U.S. Caribbean (Kendall et al., 2001) for these areas. Updated shallow-water benthic habitat maps for the Guánica/Parguera region are timely in light of ongoing restoration efforts in the Guánica Bay watershed. The bay is served directly by one river, the Rio Loco, which flows intermittently and more frequently during the rainy season. The watershed has gone through a series of manipulations and alterations in past decades, mainly associated with agricultural practices, including irrigation systems, in the upper watershed. The Guánica Lagoon, previously situated to the north of the bay, was historically the largest freshwater lagoon in Puerto Rico and served as a natural filter and sediment sink prior to the discharge of the Rio Loco into the Bay. Following alterations by the Southwest Water Project in the 1950s, the Lagoon’s adjacent wetland system was ditched and drained; no longer filtering and trapping sediment from the Rio Loco. Land use in the Guánica Bay/Rio Loco watershed has also gone through several changes (CWP, 2008). Similar to much of Puerto Rico, the area was largely deforested for sugar cane cultivation in the 1800s, although reforestation of some areas occurred following the cessation of sugar cane production (Warne et al., 2005). The northern area of the watershed is generally mountainous and is characterized by a mix of forested and agricultural lands, particularly coffee plantations. Closer to the coast, the Lajas Valley Agricultural Reserve extends north of Guánica Bay to the southwest corner of the island. The land use practices and watershed changes outlined above have resulted in large amounts of sediment being distributed in the Rio Loco river valley (CWP, 2008). Storm events and seasonal flooding also transport large amounts of sediment to the coastal waters. The threats of upstream watershed practices to coral reefs and the nearshore marine environment have been gaining recognition. Guánica Bay, and the adjacent marine waters, has been identified as a “management priority area” by NOAA’s Coral Reef Conservation Program (CRCP, 2012). In a recent Guánica Bay watershed management plan, several critical issues were outlined in regards to land-based sources of pollution (LBSP; CWP, 2008). These include: upland erosion from coffee agriculture, filling of reservoirs with sediment, in-stream channel erosion, loss of historical Guánica lagoon, legacy contaminants and sewage treatment (CWP, 2008). The plan recommended several management actions that could be taken to reduce impacts of LBSP, which form the basis of Guánica watershed restoration efforts

Baseline assessment of Guánica Bay, Puerto Rico in support of watershed restoration

Guánica Bay is a major estuary on the southwest coast of Puerto Rico. Significant coral reef ecosystems are present outside the bay. These valuable habitats may be impacted by transport of sediments, nutrients and contaminants from the watershed, through the bay and into the offshore waters. The National Oceanic and Atmospheric Administration’s (NOAA) National Centers for Coastal Ocean Science (NCCOS), in consultation with local and regional experts, conducted an interdisciplinary assessment of coral reef ecosystems, contaminants, sedimentation rates and nutrient distribution patterns in and around Guánica Bay. This work was conducted using many of the same protocols as ongoing monitoring work underway elsewhere in the U.S. Caribbean and has enabled comparisons among coral reef ecosystems between this study and other locations in the region. This characterization of Guánica marine ecosystems establishes benchmark conditions that can be used for comparative documentation of future change, including possible negative outcomes due to future land use change, or improvement in environmental conditions arising from management actions. This report is organized into six chapters that represent a suite of interrelated studies. Chapter 1 provides a short introduction to the study area. Chapter 2 is focused on biogeographic assessments and benthic mapping of the study area, including new surveys of fish, marine debris and reef communities on hardbottom habitats in the study area. Chapter 3 quantifies the distribution and magnitude of a suite of contaminants (e.g., heavy metals, PAHs, PCBs, pesticides) in both surface sediments and coral tissues. Chapter 4 presents results of sedimentation measurements in and outside of the bay. Chapter 5 examines the distribution of nutrients in in the bay, offshore from the bay and in the watershed. Chapter 6 is a brief summary discussion that highlights key findings of the entire suite of studies

The truth about metagenomics: quantifying and counteracting bias in 16S rRNA studies

Background Characterizing microbial communities via next-generation sequencing is subject to a number of pitfalls involving sample processing. The observed community composition can be a severe distortion of the quantities of bacteria actually present in the microbiome, hampering analysis and threatening the validity of conclusions from metagenomic studies. We introduce an experimental protocol using mock communities for quantifying and characterizing bias introduced in the sample processing pipeline. We used 80 bacterial mock communities comprised of prescribed proportions of cells from seven vaginally-relevant bacterial strains to assess the bias introduced in the sample processing pipeline. We created two additional sets of 80 mock communities by mixing prescribed quantities of DNA and PCR product to quantify the relative contribution to bias of (1) DNA extraction, (2) PCR amplification, and (3) sequencing and taxonomic classification for particular choices of protocols for each step. We developed models to predict the “true” composition of environmental samples based on the observed proportions, and applied them to a set of clinical vaginal samples from a single subject during four visits. Results We observed that using different DNA extraction kits can produce dramatically different results but bias is introduced regardless of the choice of kit. We observed error rates from bias of over 85% in some samples, while technical variation was very low at less than 5% for most bacteria. The effects of DNA extraction and PCR amplification for our protocols were much larger than those due to sequencing and classification. The processing steps affected different bacteria in different ways, resulting in amplified and suppressed observed proportions of a community. When predictive models were applied to clinical samples from a subject, the predicted microbiome profiles were better reflections of the physiology and diagnosis of the subject at the visits than the observed community compositions. Conclusions Bias in 16S studies due to DNA extraction and PCR amplification will continue to require attention despite further advances in sequencing technology. Analysis of mock communities can help assess bias and facilitate the interpretation of results from environmental samples

Mobilizing Community based Organizations to Address Obesity: Our Communities, Our Solutions

Purpose: The purpose of the focus groups is to: 1) explore existing thought patterns and lived experiences about eating habits and daily activity of African American (AA) children at risk for childhood obesity, 2) acquire an in-depth explanation of participants’ attitudes towards factors related to childhood obesity encompassing their social norms, dietary habits, and environmental influences, and 3) explore rationales for differences between children and parental perspectives of ideal health. Method: Using a Community Based Participatory Research (CPBR) framework, the HAUL research team, comprised of public health professionals, local policy makers, physicians, and community leaders, will facilitate 20 focus groups (10 among children and 10 among their parents) with community members accessing HAUL services. Focus group results will be used to develop a larger population based survey to further investigate obesity related risk factors. Use of Geographic Information System (GIS) mapping will identify resources (parks, fitness centers, healthy food markets, health clinics, etc.) in the residential communities of the target population. Results: At the time of submission, 6 focus groups have been completed (28 participants, 14 children, 14 adults). The primary themes identifying differences in responses of parents and children address proximity of access to healthy food choices, food type consumed by children regularly, and children’s preference for fast food versus home cooked meals. These themes will guide the development of the “UT-HAUL Healthy Houston Initiative”, a pilot program seeking to increase healthy decision making and access to wellness services among AAs. Conclusions: “UT-HAUL Healthy Houston Initiative” is the initial course of action in addressing societal factors, renowned for breeding social injustice commonplace in AA populations. Our goal is to expand the Urban League movement\u27s efficacy by identifying data that will combat social injustices while strengthening the core of the communities we serve

A new mild hyperthermia device to treat vascular involvement in cancer surgery

Abstract Surgical margin status in cancer surgery represents an important oncologic parameter affecting overall prognosis. The risk of disease recurrence is minimized and survival often prolonged if margin-negative resection can be accomplished during cancer surgery. Unfortunately, negative margins are not always surgically achievable due to tumor invasion into adjacent tissues or involvement of critical vasculature. Herein, we present a novel intra-operative device created to facilitate a uniform and mild heating profile to cause hyperthermic destruction of vessel-encasing tumors while safeguarding the encased vessel. We use pancreatic ductal adenocarcinoma as an in vitro and an in vivo cancer model for these studies as it is a representative model of a tumor that commonly involves major mesenteric vessels. In vitro data suggests that mild hyperthermia (41–46 °C for ten minutes) is an optimal thermal dose to induce high levels of cancer cell death, alter cancer cell’s proteomic profiles and eliminate cancer stem cells while preserving non-malignant cells. In vivo and in silico data supports the well-known phenomena of a vascular heat sink effect that causes high temperature differentials through tissues undergoing hyperthermia, however temperatures can be predicted and used as a tool for the surgeon to adjust thermal doses delivered for various tumor margins

PET Imaging of Extracellular pH in Tumors with \u3csup\u3e64\u3c/sup\u3eCu- and \u3csup\u3e18\u3c/sup\u3eF-Labeled pHLIP Peptides: A Structure–Activity Optimization Study

pH (low) insertion peptides (pHLIP peptides) target acidic extracellular environments in vivo due to pH-dependent cellular membrane insertion. Two variants (Var3 and Var7) and wild-type (WT) pHLIP peptides have shown promise for in vivo imaging of breast cancer. Two positron emitting radionuclides (64Cu and 18F) were used to label the NOTA- and NO2A-derivatized Var3, Var7, and WT peptides for in vivo biodistribution studies in 4T1 orthotopic tumor-bearing BALB/c mice. All of the constructs were radiolabeled with 64Cu or [18F]-AlF in good yield. The in vivo biodistribution of the 12 constructs in 4T1 orthotopic allografted female BALB/c mice indicated that NO2A-cysVar3, radiolabeled with either 18F (4T1 uptake; 8.9 ± 1.7%ID/g at 4 h p.i.) or 64Cu (4T1 uptake; 8.2 ± 0.9%ID/g at 4 h p.i. and 19.2 ± 1.8% ID/g at 24 h p.i.), shows the most promise for clinical translation. Additional studies to investigate other tumor models (melanoma, prostate, and brain tumor models) indicated the universality of tumor targeting of these tracers. From this study, future clinical translation will focus on 18F- or 64Cu-labeled NO2A-cysVar3

Toward a chemical reanalysis in a coupled chemistry-climate model: an evaluation of MOPITT CO assimilation and its impact on tropospheric composition

We examine in detail a 1 year global reanalysis of carbon monoxide (CO) that is based on joint assimilation of conventional meteorological observations and Measurement of Pollution in The Troposphere (MOPITT) multispectral CO retrievals in the Community Earth System Model (CESM). Our focus is to assess the impact to the chemical system when CO distribution is constrained in a coupled full chemistry-climate model like CESM. To do this, we first evaluate the joint reanalysis (MOPITT Reanalysis) against four sets of independent observations and compare its performance against a reanalysis with no MOPITT assimilation (Control Run). We then investigate the CO burden and chemical response with the aid of tagged sectoral CO tracers. We estimate the total tropospheric CO burden in 2002 (from ensemble mean and spread) to be 371 ± 12% Tg for MOPITT Reanalysis and 291 ± 9% Tg for Control Run. Our multispecies analysis of this difference suggests that (a) direct emissions of CO and hydrocarbons are too low in the inventory used in this study and (b) chemical oxidation, transport, and deposition processes are not accurately and consistently represented in the model. Increases in CO led to net reduction of OH and subsequent longer lifetime of CH4 (Control Run: 8.7 years versus MOPITT Reanalysis: 9.3 years). Yet at the same time, this increase led to 5-10% enhancement of Northern Hemisphere O3 and overall photochemical activity via HOx recycling. Such nonlinear effects further complicate the attribution to uncertainties in direct emissions alone. This has implications to chemistry-climate modeling and inversion studies of longer-lived species