No abstract

The changes to soil properties due to biochar addition may affect both the direction and magnitude of priming effects. However, the mechanisms involved in biochar induced priming effects still remain largely unknown due to the limitation of methods to separate more than two carbon (C) sources (e.g. soil, biochar, substrate). We combined 14 C labeling with 13 C natural abundance to separate the total CO 2 from i) native soil organic C (SOC, C 3 signature), ii) added glucose (14 C labeled) and iii) biochar (C 4 signature).The primed soil CO 2 emissions following a large addition of glucose (1000 mg glucose kg-1 soil) to one Chinese and one German Luvisol soil were much larger (140% and 53% respectively) in a soil recently amended with maize derived biochar (pyrolyzed at 400°C), compared to non amended soil. Glucose addition at a lower rate (100 mg C kg-1 soil) produced no significant differences in priming effects of native soil organic matter between the biochar amended and non-amended soils. Glucose also caused priming of biochar decomposition, with an additional C 4 biochar loss of between 270 µg CO 2-C g-1 and 540 µg CO 2-C g-1 depending on soils and glucose concentrations. Approaches using two stable isotopes (13 C and 12 C) have previously been limited to partitioning two sources (biochar C and soil organic C). Here, for the first time, 14 C labeling was combined with 13 C natural abundance to partition three C sources in a biochar amended soil. By partitioning soil CO 2 emissions derived from SOC, from added biochar and glucose decompositions, this study provides a better understanding of the priming effects following addition of substrates to biochar amended soil, to approximate to the true complexity of biochar enriched soils.

Objective: To evaluate the mechanical property and biocompatibility of the Wnt pathway inhibitor (ICG-001) delivering collagen/poly(l-lactide-co-caprolactone) (P(LLA-CL)) scaffold for urethroplasty, and also the feasibility of inhibiting the extracellular matrix (ECM) expression in vitro and in vivo. Methods: ICG-001 (1 mg (2 mM)) was loaded into a (P(LLA-CL)) scaffold with the co-axial electrospinning technique. The characteristics of the mechanical property and drug release fashion of scaffolds were tested with a mechanical testing machine (Instron) and high-performance liquid chromatography (HPLC). Rabbit bladder epithelial cells and the dermal fibroblasts were isolated by enzymatic digestion method. (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay) and scanning electron microscopy (SEM) were used to evaluate the viability and proliferation of the cells on the scaffolds. Fibrolasts treated with TGF-β1 and ICG-001 released medium from scaffolds were used to evaluate the anti-fibrosis effect through immunofluorescence, real time PCR and western blot. Urethrography and histology were used to evaluate the efficacy of urethral implantation. Results: The scaffold delivering ICG-001 was fabricated, the fiber diameter and mechanical strength of scaffolds with inhibitor were comparable with the non-drug scaffold. The SEM and MTT assay showed no toxic effect of ICG-001 to the proliferation of epithelial cells on the collagen/P(LLA-CL) scaffold with ICG-001. After treatment with culture medium released from the drug-delivering scaffold, the expression of Collagen type 1, 3 and fibronectin of fibroblasts could be inhibited significantly at the mRNA and protein levels. In the results of urethrography, urethral strictures and fistulas were found in the rabbits treated with non-ICG-001 delivering scaffolds, but all the rabbits treated with ICG-001-delivering scaffolds showed wide caliber in urethras. Histology results showed less collagen but more smooth muscle and thicker epithelium in urethras repaired with ICG-001 delivering scaffolds. Conclusion: After loading with the Wnt signal pathway inhibitor ICG-001, the Collagen/P(LLA-CL) scaffold could facilitate a decrease in the ECM deposition of fibroblasts. The ICG-001 delivering Collagen/P(LLA-CL) nanofibrous scaffold seeded with epithelial cells has the potential to be a promising substitute material for urethroplasty. Longer follow-up study in larger animals is needed in the future.

Urethral strictures remain a reconstructive challenge, due to less than satisfactory outcomes and high incidence of stricture recurrence. An “ideal” urethral reconstruction should establish similar architecture and function as the original urethral wall. We fabricated a novel tissue-engineered bionic urethras using cell sheet technology and report their viability in a canine model. Small amounts of oral and adipose tissues were harvested, and adipose-derived stem cells, oral mucosal epithelial cells, and oral mucosal fibroblasts were isolated and used to prepare cell sheets. The cell sheets were hierarchically tubularized to form 3-layer tissue-engineered urethras and labeled by ultrasmall super-paramagnetic iron oxide (USPIO). The constructed tissue-engineered urethras were transplanted subcutaneously for 3 weeks to promote the revascularization and biomechanical strength of the implant. Then, 2 cm length of the tubularized penile urethra was replaced by tissue-engineered bionic urethra. At 3 months of urethral replacement, USPIO-labeled tissue-engineered bionic urethra can be effectively detected by MRI at the transplant site. Histologically, the retrieved bionic urethras still displayed 3 layers, including an epithelial layer, a fibrous layer, and a myoblast layer. Three weeks after subcutaneous transplantation, immunofluorescence analysis showed the density of blood vessels in bionic urethra was significantly increased following the initial establishment of the constructs and was further up-regulated at 3 months after urethral replacement and was close to normal level in urethral tissue. Our study is the first to experimentally demonstrate 3-layer tissue-engineered urethras can be established using cell sheet technology and can promote the regeneration of structural and functional urethras similar to normal urethra.

Recent studies have shown that M. elongata (M. elongata) isolated from Populus field sites has a dual endophyte–saprotroph lifestyle and is able to promote the growth of Populus. However, little is known about the host fidelity of M. elongata and whether M. elongata strains differ from one another in their ability to promote plant growth. Here, we compared the impacts of three Populus-associated M. elongata isolates (PMI 77, PMI 93, and PMI 624) on the growth of seven different crop species by measuring plant height, plant dry biomass, and leaf area. M. elongata isolates PMI 624 and PMI 93 increased the plant height, leaf area, and plant dry weight of Citrullus lanatus, Zea mays, Solanum lycopersicum, and Cucurbita to a much greater degree than PMI 77 (33.9% to 14.1%). No significant impacts were observed for any isolate on the growth of Abelmoschus esculentus or Glycine max. On the contrary, Glycine max significantly decreased in height by 30.6% after the inoculation of M. elongata PMI 77. In conclusion, this study demonstrates that M. elongata generally promoted metrics of the plant performance among a diverse set of importantly non-leguminous crop species. Future research on understanding the molecular mechanisms that underlie strain and host variability is warranted.