Activation of formate hydrogen-lyase via expression of uptake [NiFe]-hydrogenase in Escherichia coli BL21(DE3)

Background: Several recent studies have reported successful hydrogen (H-2) production achieved via recombinant expression of uptake [NiFe]-hydrogenases from Hydrogenovibrio marinus, Rhodobacter sphaeroides, and Escherichia coli (hydrogenase-1) in E. coli BL21(DE3), a strain that lacks H-2-evolving activity. However, there are some unclear points that do not support the conclusion that the recombinant hydrogenases are responsible for the in vivo H-2 production. Results: Unlike wild-type BL21(DE3), the recombinant BL21(DE3) strains possessed formate hydrogen-lyase (FHL) activities. Through experiments using fdhF (formate dehydrogenase-H) or hycE (hydrogenase-3) mutants, it was shown that H-2 production was almost exclusively dependent on FHL. Upon expression of hydrogenase, extracellular formate concentration was changed even in the mutant strains lacking FHL, indicating that formate metabolism other than FHL was also affected. The two subunits of H. marinus uptake [NiFe]-hydrogenase could activate FHL independently of each other, implying the presence of more than two different mechanisms for FHL activation in BL21(DE3). It was also revealed that the signal peptide in the small subunit was essential for activation of FHL via the small subunit. Conclusions: Herein, we demonstrated that the production of H-2 was indeed induced via native FHL activated by the expression of recombinant hydrogenases. The recombinant strains with [NiFe]-hydrogenase appear to be unsuitable for practical in vivo H-2 production due to their relatively low H-2 yields and productivities. We suggest that an improved H-2-producing cell factory could be designed by constructing a well characterized and overproduced synthetic H-2 pathway and fully activating the native FHL in BL21(DE3).1122Ysciescopu

Engineering de novo disulfide bond in bacterial alpha-type carbonic anhydrase for thermostable carbon sequestration

Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial alpha-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 degrees C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions.1194Ysciescopu

Diversity, floristic composition, and structure of the woody vegetation of the Cerrado in the Cerrado–Amazon transition zone in Mato Grosso, Brazil

The final publication is available at Springer via http://dx.doi.org/10.1007/s40415-015-0186-2We compared the diversity and species composition and the structure of the vegetation of three distinct Cerrado phytophysiognomies (Cerradão, Dense Cerrado, and Typical Cerrado) in the Cerrado–Amazon transition, Mato Grosso (Brazil). Species richness (observed and estimated) in the Cerradão and Dense Cerrado was higher than that recorded in the Typical Cerrado. Species diversity, based on a Rényi profile, was highest in the Dense Cerrado, in comparison with the other phytophysiognomies. We recorded a higher number of exclusive species in the Cerradão and a greater similarity (Morisita and Sørensen indices) between this vegetation type and the Dense Cerrado. While individuals were tallest in the Cerradão and Dense Cerrado and lowest in the Typical Cerrado, there was no difference among phytophysiognomies in mean diameter. A gradient in decreasing species richness and diversity (hypothesis 1) and vegetation vertical structure (hypothesis 3) was expected for the Cerradão–Dense Cerrado–Typical Cerrado; however, neither hypothesis was supported by the results. The Cerradão and Dense Cerrado were most similar in species composition not confirming hypothesis 2, which predicted that the two savanna vegetation types (Dense Cerrado and Typical Cerrado) were more similar to one another than either is to the woodland (Cerradão). Overall, the similarities among the three study communities depended on the type of parameter analyzed. While the species richness and the vertical and structure of the vegetation of the Cerradão and Dense Cerrado are closely similar, the Cerradão and Typical Cerrado are more similar in their species diversity. With regard to the floristic composition, Dense Cerrado occupies an intermediate position between Cerradão and Typical Cerrado.UNEMAT Graduate Program in Ecology and ConservationBrazilian Higher Education Training Program (CAPES)“Tropical Biomes in Transition – TROBIT”CAPES/Science without Borders ProgramPELD/CNPq (Long-Term Ecological Studies)PROCAD UnB/UNEMA

Yield gaps, nutrient use efficiencies and response to fertilisers by maize across heterogeneous smallholder farms of western Kenya

The need to promote fertiliser use by African smallholder farmers to counteract the current decline in per capita food production is widely recognised. But soil heterogeneity results in variable responses of crops to fertilisers within single farms. We used existing databases on maize production under farmer (F-M) and researcher management (R-M) to analyse the effect of soil heterogeneity on the different components of nutrient use efficiency by maize growing on smallholder farms in western Kenya: nutrient availability, capture and conversion efficiencies and crop biomass partitioning. Subsequently, we used the simple model QUEFTS to calculate nutrient recovery efficiencies from the R-M plots and to calculate attainable yields with and without fertilisers based on measured soil properties across heterogeneous farms. The yield gap of maize between F-M and R-M varied from 0.5 to 3 t grain ha-1 season-1 across field types and localities. Poor fields under R-M yielded better than F-M, even without fertilisers. Such differences, of up to 1.1 t ha-1 greater yields under R-M conditions are attributable to improved agronomic management and germplasm. The relative response of maize to N-P-K fertilisers tended to decrease with increasing soil quality (soil C and extractable P), from a maximum of 4.4-fold to -0.5- fold relative to the control. Soil heterogeneity affected resource use efficiencies mainly through effects on the efficiency of resource capture. Apparent recovery efficiencies varied between 0 and 70% for N, 0 and 15% for P, and 0 to 52% for K. Resource conversion efficiencies were less variable across fields and localities, with average values of 97 kg DM kg-1 N, 558 kg DM kg-1 P and 111 kg DM kg-1 K taken up. Using measured soil chemical properties QUEFTS over-estimated observed yields under F-M, indicating that variable crop performance within and across farms cannot be ascribed solely to soil nutrient availability. For the R-M plots QUEFTS predicted positive crop responses to application of 30 kg P ha-1 and 30 kg P ha-1 + 90 kg N ha-1 for a wide range of soil qualities, indicating that there is room to improve current crop productivity through fertiliser use. To ensure their efficient use in sub-Saharan Africa mineral fertilisers should be: (1) targeted to specific niches of soil fertility within heterogeneous farms; and (2) go hand-in-hand with the implementation of agronomic measures to improve their capture and utilisation

Cognitive and behavioral predictors of light therapy use

Objective: Although light therapy is effective in the treatment of seasonal affective disorder (SAD) and other mood disorders, only 53-79% of individuals with SAD meet remission criteria after light therapy. Perhaps more importantly, only 12-41% of individuals with SAD continue to use the treatment even after a previous winter of successful treatment. Method: Participants completed surveys regarding (1) social, cognitive, and behavioral variables used to evaluate treatment adherence for other health-related issues, expectations and credibility of light therapy, (2) a depression symptoms scale, and (3) self-reported light therapy use. Results: Individuals age 18 or older responded (n = 40), all reporting having been diagnosed with a mood disorder for which light therapy is indicated. Social support and self-efficacy scores were predictive of light therapy use (p's<.05). Conclusion: The findings suggest that testing social support and self-efficacy in a diagnosed patient population may identify factors related to the decision to use light therapy. Treatments that impact social support and self-efficacy may improve treatment response to light therapy in SAD. © 2012 Roecklein et al

A randomised controlled trial and cost-effectiveness evaluation of "booster" interventions to sustain increases in physical activity in middle-aged adults in deprived urban neighbourhoods

Background: Systematic reviews have identified a range of brief interventions which increase physical activity in previously sedentary people. There is an absence of evidence about whether follow up beyond three months can maintain long term physical activity. This study assesses whether it is worth providing motivational interviews, three months after giving initial advice, to those who have become more active. Methods/Design: Study candidates (n = 1500) will initially be given an interactive DVD and receive two telephone follow ups at monthly intervals checking on receipt and use of the DVD. Only those that have increased their physical activity after three months (n = 600) will be randomised into the study. These participants will receive either a "mini booster" (n = 200), "full booster" (n = 200) or no booster (n = 200). The "mini booster" consists of two telephone calls one month apart to discuss physical activity and maintenance strategies. The "full booster" consists of a face-to-face meeting with the facilitator at the same intervals. The purpose of these booster sessions is to help the individual maintain their increase in physical activity. Differences in physical activity, quality of life and costs associated with the booster interventions, will be measured three and nine months from randomisation. The research will be conducted in 20 of the most deprived neighbourhoods in Sheffield, which have large, ethnically diverse populations, high levels of economic deprivation, low levels of physical activity, poorer health and shorter life expectancy. Participants will be recruited through general practices and community groups, as well as by postal invitation, to ensure the participation of minority ethnic groups and those with lower levels of literacy. Sheffield City Council and Primary Care Trust fund a range of facilities and activities to promote physical activity and variations in access to these between neighbourhoods will make it possible to examine whether the effectiveness of the intervention is modified by access to community facilities. A one-year integrated feasibility study will confirm that recruitment targets are achievable based on a 10% sample.Discussion: The choice of study population, study interventions, brief intervention preceding the study, and outcome measure are discussed

Microfluidic impedance biosensors for monitoring a single and multiple cancer cells in anticancer drug treatments

In this work, we present a novel microfluidic impedance biosensor chip for trapping both a single and multiple cancer cells and monitoring their response to the anti-cancer drug treatment. By designing different sizes of working microelectrodes together with the V-shaped cell capture structures, a single or multiple cells are trapped on the microelectrodes surfaces. In addition, by utilizing the passive pumping method, cells can be trapped and positioned inside the microchannels without the need of using the outer micro pump or syringe. The impedance change induced by the response of cells to the anticancer drug Cisplatin treatment was successfully recorded. The proposed biosensor chip has a great potential for applications in cancer cell research, drug screening, and quantification of cancer cells from various tumor stages. The results of this study open potential research collaborations about development of cost-effective devices and lab-on-chips for early disease detection, studies of cancerous cells and their response to anti-cancer drugs to optimize cancer treatments, characterisation of mechanical properties of cells, new drug delivery mechanisms, and micro and nano manufacturing

Combination treatment with doxorubicin and gamitrinib synergistically augments anticancer activity through enhanced activation of Bim

Background: A common approach to cancer therapy in clinical practice is the combination of several drugs to boost the anticancer activity of available drugs while suppressing their unwanted side effects. In this regard, we examined the efficacy of combination treatment with the widely-used genotoxic drug doxorubicin and the mitochondriotoxic Hsp90 inhibitor gamitrinib to exploit disparate stress signaling pathways for cancer therapy.Methods: The cytotoxicity of the drugs as single agents or in combination against several cancer cell types was analyzed by MTT assay and the synergism of the drug combination was evaluated by calculating the combination index. To understand the molecular mechanism of the drug synergism, stress signaling pathways were analyzed after drug combination. Two xenograft models with breast and prostate cancer cells were used to evaluate anticancer activity of the drug combination in vivo. Cardiotoxicity was assessed by tissue histology and serum creatine phosphokinase concentration.Results: Gamitrinib sensitized various human cancer cells to doxorubicin treatment, and combination treatment with the two drugs synergistically increased apoptosis. The cytotoxicity of the drug combination involved activation and mitochondrial accumulation of the proapoptotic Bcl-2 family member Bim. Activation of Bim was associated with increased expression of the proapoptotic transcription factor C/EBP-homologous protein and enhanced activation of the stress kinase c-Jun N-terminal kinase. Combined drug treatment with doxorubicin and gamitrinib dramatically reduced in vivo tumor growth in prostate and breast xenograft models without increasing cardiotoxicity.Conclusions: The drug combination showed synergistic anticancer activities toward various cancer cells without aggravating the cardiotoxic side effects of doxorubicin, suggesting that the full therapeutic potential of doxorubicin can be unleashed through combination with gamitrinib.open