Production of indigo by recombinant bacteria

Indigo is an economically important dye, especially for the textile industry and the dyeing of denim fabrics for jeans and garments. Around 80,000 tonnes of indigo are chemically produced each year with the use of non-renewable petrochemicals and the use and generation of toxic compounds. As many microorganisms and their enzymes are able to synthesise indigo after the expression of specific oxygenases and hydroxylases, microbial fermentation could offer a more sustainable and environmentally friendly manufacturing platform. Although multiple small-scale studies have been performed, several existing research gaps still hinder the effective translation of these biochemical approaches. No article has evaluated the feasibility and relevance of the current understanding and development of indigo biocatalysis for real-life industrial applications. There is no record of either established or practically tested large-scale bioprocess for the biosynthesis of indigo. To address this, upstream and downstream processing considerations were carried out for indigo biosynthesis. 5 classes of potential biocatalysts were identified, and 2 possible bioprocess flowsheets were designed that facilitate generating either a pre-reduced dye solution or a dry powder product. Furthermore, considering the publicly available data on the development of relevant technology and common bioprocess facilities, possible platform and process values were estimated, including titre, DSP yield, potential plant capacities, fermenter size and batch schedule. This allowed us to project the realistic annual output of a potential indigo biosynthesis platform as 540 tonnes. This was interpreted as an industrially relevant quantity, sufficient to provide an annual dye supply to a single industrial-size denim dyeing plant. The conducted sensitivity analysis showed that this anticipated output is most sensitive to changes in the reaction titer, which can bring a 27.8% increase or a 94.4% drop. Thus, although such a biological platform would require careful consideration, fine-tuning and optimization before real-life implementation, the recombinant indigo biosynthesis was found as already attractive for business exploitation for both, luxury segment customers and mass-producers of denim garments

Pasos Hacia La Salud: a randomized controlled trial of an internet-delivered physical activity intervention for Latinas.

BackgroundInternet access has grown markedly in Latinos during the past decade. However, there have been no Internet-based physical activity interventions designed for Latinos, despite large disparities in lifestyle-related conditions, such as obesity and diabetes, particularly in Latina women. The current study tested the efficacy of a 6-month culturally adapted, individually tailored, Spanish-language Internet-based physical activity intervention.MethodsInactive Latinas (N = 205) were randomly assigned to the Tailored Physical Activity Internet Intervention or the Wellness Contact Control Internet Group. Participants in both groups received emails on a tapered schedule over 6 months to alert them to new content on the website. The primary outcome was minutes/week of moderate to vigorous physical activity (MVPA) at 6 months as measured by the 7-Day Physical Activity Recall; activity was also measured by accelerometers. Data were collected between 2011 and 2014 and analyzed in 2015 at the University of California, San Diego.ResultsIncreases in minutes/week of MVPA were significantly greater in the Intervention Group compared to the Control Group (mean difference = 50.00, SE = 9.5, p < 0.01). Increases in objectively measured MVPA were also significantly larger in the Intervention Group (mean differences = 31.0, SE = 10.7, p < .01). The Intervention Group was also significantly more likely to meet national physical activity guidelines at 6 months (OR = 3.12, 95% CI 1.46-6.66, p < .05).ConclusionFindings from the current study suggest that this Internet-delivered individually tailored intervention successfully increased MVPA in Latinas compared to a Wellness Contact Control Internet Group.Trial registrationNCT01834287

Physical activity maintenance among Spanish-speaking Latinas in a randomized controlled trial of an Internet-based intervention.

Spanish-speaking Latinas have some of the lowest rates of meeting physical activity guidelines in the U.S. and are at high risk for many related chronic diseases. The purpose of the current study was to examine the maintenance of a culturally and individually-tailored Internet-based physical activity intervention for Spanish-speaking Latinas. Inactive Latinas (N  =  205) were randomly assigned to a 6-month Tailored Physical Activity Internet Intervention or a Wellness Contact Control Internet Group, with a 6-month follow-up. Maintenance was measured by assessing group differences in minutes per week of self-reported and accelerometer measured moderate to vigorous physical activity (MVPA) at 12 months after baseline and changes in MVPA between the end of the active intervention (month 6) and the end of the study (month 12). Potential moderators of the intervention were also examined. Data were collected between 2011 and 2014, and were analyzed in 2015 at the University of California, San Diego. The Intervention Group engaged in significantly more minutes of MVPA per week than the Control Group at the end of the maintenance period for both self-reported (mean diff. = 30.68, SE = 11.27, p = .007) and accelerometer measured (mean diff. = 11.47, SE = 3.19, p = .01) MVPA. There were no significant between- or within-group changes in MVPA from month 6 to 12. Greater intervention effects were seen for those with lower BMI (BMI × intervention = -6.67, SE = 2.88, p = .02) and lower perceived places to walk to in their neighborhood (access × intervention = -43.25, SE = 19.07, p = .02), with a trend for less family support (social support × intervention = -3.49, SE = 2.05, p = .08). Acculturation, health literacy, and physical activity related psychosocial variables were not significant moderators of the intervention effect during the maintenance period. Findings from the current study support the efficacy of an Internet-delivered individually tailored intervention for maintenance of MVPA gains over time

Mapping and Evaluating Marine Protected Areas and Ecosystem Services: A Transdisciplinary Delphi Forecasting Process Framework

Marine Protected Areas (MPAs) are an important tool for management and conservation and play an increasingly recognised role in societal and human well-being. However, the assessment of MPAs often lacks a simultaneous consideration of ecological and socio-economic outcomes, and this can lead to misconceptions on the effectiveness of MPAs. In this perspective, we present a transdisciplinary approach based on the Delphi method for mapping and evaluating Marine Protected Areas for their ability to protect biodiversity while providing Ecosystem Services (ES) and related human well-being benefits - i.e., the ecosystem outputs from which people benefit. We highlight the need to include the human dimensions of marine protection in such assessments, given that the effectiveness of MPAs over time is conditional on the social, cultural and institutional contexts in which MPAs evolve. Our approach supports Ecosystem-Based Management and highlights the importance of MPAs in achieving restoration, conservation, and sustainable development objectives in relation to EU Directives such as the Marine Strategy Framework Directive (MSFD), the Maritime Spatial Planning Directive (MSPD), and the Common Fisheries Policy (CFP)

Geological imprint of methane seepage on the seabed and biota of the convergent Hikurangi Margin, New Zealand: box core and grab carbonate results

Short box cores (to 30 cm bsf) and seafloor carbonate grab samples were acquired at mapped hydrocarbon seep sites (600–1200 m water depths) during the 2007 RV SONNE SO191 cruise on the Hikurangi Margin offshore eastern North Island, New Zealand, to evaluate the influence of methane seepage on sedimentologic, biotic, mineralogic and stable isotopic attributes of seabed sediments. Sedimentary horizons in the box cores consist of siliciclastic silts and sands, shell beds and nodular, microcrystalline aragonite bands up to 15 cm thick. The megafauna is dominated by infaunal to semi-infaunal chemosymbiotic bivalves (Calyptogena, Lucinoma, and Acharax), as well as associated worms and carnivorous and grazing gastropods. Burrows in silts, some occupied by worms or juvenile Acharax, mainly have simple morphologies more typical of high-energy, nearshore settings than deep-sea environments, while a few are large and sparsely branched with wall scratch marks inferred to be of decapod crustacean origin. The box core silts and nodular carbonate samples vary in TOC content from 0.2 to 0.9 wt.%, carbonate content from 4 to 78%, and δ13C and δ18O values from − 50.3 to − 0.6‰ PDB and + 0.77 to + 3.2‰ PDB, respectively. Low carbonate content silt samples have the most enriched δ13C values, implying a seawater source for their pore water bicarbonate. Negative δ13C and positive δ18O values typify the nodular, microcrystalline aragonite bands, indicating formation during microbially mediated, sulphate-dependent anaerobic oxidation of methane (AOM) in a cold, near-seafloor environment, as is also supported by lipid biomarker data. A clear isotopic mixing trend of decreasing δ13C and increasing δ18O and carbonate content in the fine (< 100 µm) carbonate fraction of the host silts also has been reported from other methane seep provinces, and suggests a heterogeneous influx of methane-rich see

An isoform of the giant protein titin is a master regulator of human T lymphocyte trafficking

Response to multiple microenvironmental cues and resilience to mechanical stress are essential features of trafficking leukocytes. Here, we describe unexpected role of titin (TTN), the largest protein encoded by the human genome, in the regulation of mechanisms of lymphocyte trafficking. Human T and B lymphocytes express five TTN isoforms, exhibiting cell-specific expression, distinct localization to plasma membrane microdomains, and different distribution to cytosolic versus nuclear compartments. In T lymphocytes, the LTTN1 isoform governs the morphogenesis of plasma membrane microvilli independently of ERM protein phosphorylation status, thus allowing selectin-mediated capturing and rolling adhesions. Likewise, LTTN1 controls chemokine-triggered integrin activation. Accordingly, LTTN1 mediates rho and rap small GTPases activation, but not actin polymerization. In contrast, chemotaxis is facilitated by LTTN1 degradation. Finally, LTTN1 controls resilience to passive cell deformation and ensures T lymphocyte survival in the blood stream. LTTN1 is, thus, a critical and versatile housekeeping regulator of T lymphocyte trafficking

Small intestinal mucosa expression of putative chaperone fls485

<p>Abstract</p> <p>Background</p> <p>Maturation of enterocytes along the small intestinal crypt-villus axis is associated with significant changes in gene expression profiles. <it>fls485 </it>coding a putative chaperone protein has been recently suggested as a gene involved in this process. The aim of the present study was to analyze <it>fls48</it>5 expression in human small intestinal mucosa.</p> <p>Methods</p> <p><it>fls485 </it>expression in purified normal or intestinal mucosa affected with celiac disease was investigated with a molecular approach including qRT-PCR, Western blotting, and expression strategies. Molecular data were corroborated with several <it>in situ </it>techniques and usage of newly synthesized mouse monoclonal antibodies.</p> <p>Results</p> <p>fls485 mRNA expression was preferentially found in enterocytes and chromaffine cells of human intestinal mucosa as well as in several cell lines including Rko, Lovo, and CaCo2 cells. Western blot analysis with our new anti-fls485 antibodies revealed at least two fls485 proteins. In a functional CaCo2 model, an increase in fls485 expression was paralleled by cellular maturation stage. Immunohistochemistry demonstrated fls485 as a cytosolic protein with a slightly increasing expression gradient along the crypt-villus axis which was impaired in celiac disease Marsh IIIa-c.</p> <p>Conclusions</p> <p>Expression and synthesis of fls485 are found in surface lining epithelia of normal human intestinal mucosa and deriving epithelial cell lines. An interdependence of enterocyte differentiation along the crypt-villus axis and fls485 chaperone activity might be possible.</p

Genome Sequencing of SHH Medulloblastoma Predicts Genotype-Related Response to Smoothened Inhibition

SummarySmoothened (SMO) inhibitors recently entered clinical trials for sonic-hedgehog-driven medulloblastoma (SHH-MB). Clinical response is highly variable. To understand the mechanism(s) of primary resistance and identify pathways cooperating with aberrant SHH signaling, we sequenced and profiled a large cohort of SHH-MBs (n = 133). SHH pathway mutations involved PTCH1 (across all age groups), SUFU (infants, including germline), and SMO (adults). Children >3 years old harbored an excess of downstream MYCN and GLI2 amplifications and frequent TP53 mutations, often in the germline, all of which were rare in infants and adults. Functional assays in different SHH-MB xenograft models demonstrated that SHH-MBs harboring a PTCH1 mutation were responsive to SMO inhibition, whereas tumors harboring an SUFU mutation or MYCN amplification were primarily resistant