Structure and flow calculation of cake layer on microfiltration membranes.

Submerged membrane bioreactors (SMBR) are widely used in wastewater treatment. The permeability of a membrane declines rapidly because of the formation of a cake layer on the membrane surface. In this paper, a multiple staining protocol was conducted to probe the four major foulants in the cake layer formed on a filtration membrane. Fluorescent images of the foulants were obtained using a confocal laser scanning microscope (CLSM). The three dimensional structure of the cake layer was reconstructed, and the internal flow was calculated using computational fluid dynamics (CFD). Simulation results agreed well with the experimental data on the permeability of the cake layer during filtration and showed better accuracy than the calculation by Kozeny-Carman method. ß-d-Glucopyranose polysaccharides and proteins are the two main foulants with relatively large volume fractions, while α-d-glucopyranose polysaccharides and nucleic acids have relatively large specific surface areas. The fast growth of ß-d-glucopyranose polysaccharides in the volume fraction is mainly responsible for the increase in cake volume fraction and the decrease in permeability. The specific area, or the aggregation/dispersion of foulants, is less important to its permeability compared to its volume fraction.

The Interactions Between Aligned Poly(L-Lactic Acid) Nanofibers and SH-SY5Y Cells In Vitro.

Aligned nanofibers have been regarded as promising nanomaterials in facilitating nerve regeneration. Investigating the interactions between aligned nanofibers and neuronal cells will be critically important for the design and application of aligned nanofibers in nerve tissue engineering. In this study, we explored the effects of electrospun Poly(L-Lactic Acid) (PLLA) aligned nanofibers on SH-SY5Y cells (a type of human neuroblastoma cell line) and specifically focused on the role of integrin in the PLLA aligned nanofiber-SH-SY5Y cell interaction. We found that PLLA aligned nanofibers could significantly guide the neurite outgrowth of SH-SY5Y cell, and promote the viability, proliferation, glucose and lactic acid metabolism of SH-SY5Y cell. This promotion effect could be alleviated when the functions of integrins on the SH-SY5Y cell membrane were hampered by pentapeptide GRGDS. Moreover, we found that PLLA aligned nanofibers could enhance the expression of phosphorylated-ERK1/2 (p-ERK1/2) in the SH-SY5Y cells and blocking the integrins would decrease p-ERK1/2 expression. These results suggested that PLLA aligned nanofibers might affect many cellular behaviors of SH-SY5Y cells via integrin mediated ERK pathway. Our findings provided more insights for understanding the interaction between aligned nanofibers and neuronal cells.

A Bioinspired Ultratough Composite Produced by Integration of Inorganic Ionic Oligomers within Polymer Networks.

The nacre-inspired laminates are promising materials for their excellent mechanics. However, the interfacial defects between organic-inorganic phases commonly lead to the crack propagation and fracture failure of these materials under stress. A natural biomineral, bone, has much higher bending toughness than the nacre. The small size of inorganic building units in bone improves the organic-inorganic interaction, which optimizes the material toughness. Inspired by these biological structures, here, an ultratough nanocomposite laminate is prepared by the integration of ultrasmall calcium phosphate oligomers (CPO, 1 nm in diameter) within poly(vinyl alcohol) (PVA) and sodium alginate (Alg) networks through a simple three-step strategy. Owing to the small size of inorganic building units, strong multiple molecular interactions within integrated organic-inorganic hierarchical structure are built. The resulting laminates exhibit ultrahigh bending strain (>50% without fracture) and toughness (21.5-31.0 MJ m-3), which surpass natural nacre and almost all of the synthetic laminate materials that have been reported so far. Moreover, the mechanics of this laminate is tunable by changing the water content within the bulk structure. This work provides a way for the development of organic-inorganic nanocomposites with ultrahigh bending toughness by using inorganic ionic oligomers, which can be useful in the fields of tough protective materials and energy absorbing materials.

Genome sequencing identified a novel exonic microdeletion in the RUNX2 gene that causes cleidocranial dysplasia.

BACKGROUND AND AIMS: Cleidocranial dysplasia (CCD) represents a rare autosomal dominant skeletal dysplasia caused by mutations that induce haploinsufficiency in RUNX2, the important transcription factor of osteoblasts related to bone/cartilage development and maintenance. Clavicular hypoplasia, which involves aberrant tooth/craniofacial bone/skeletal formation, is a feature of classic CCD. RUNX2 mutations can be found in approximately 60-70% of patients with CCD, and around ∼10% of these mutations are microdeletions. The present paper describes the radiological and clinical characteristics of a 5-year-old girl who showed representative CCD features, including extra teeth, aplasia of clavicles, sloping shoulders, marked calvarial hypomineralization, and osteoporosis. MATERIALS AND METHODS: We obtained genomic DNA of her family members and performed whole-genome sequencing (WGS) for samples collected from the proband. Quantitative fluorescent PCR (QF-PCR) and specific PCR plus electrophoresis were then performed as validation assays for all participants. In vitro analysis was performed. Luciferase assay for Runx2 transcription activity and evaluation of mRNA levels of Runx2 downstream osteogenic markers were conducted. RESULTS: WGS identified a 11.38-kb microdeletion in RUNX2 comprising 8-9 exons, which was validated by QF-PCR and specific PCR plus electrophoresis. In vitro experiments confirmed the pathogenicity of this variation. CONCLUSION: The present study identified a 11.38-kb microdeletion in RUNX2 that causes CCD. The deletion in the PST domain of RUNX2 reduces its transcription activity and reduces osteogenic marker levels, eventually decreasing the differentiation of osteoblasts. These findings clarify the role of the CCD-related mechanism in the development of CCD and suggest that it is important to consider copy number variation for the suspected familial patients early.

Cemented K-wire fixation for the treatment of shaft fractures of middle phalanges.

BACKGROUND: The objective of this report is to introduce an external-fixation technique using the combination of K-wires and cement. METHODS: From February 2009 to January 2015, 51 patients with shaft fractures of middle phalanges were treated with cemented K-wire fixation. The mean age of patients at surgery was 41 years. The mean time interval from injury to operation was 6±5.78days. Injured digits included index (n=18), long (n=15), ring (n=7), and little (n=11) fingers. Types of fractures were transversal (n=32), short oblique or spiral (n=5), and comminuted (n=14) fractures. Active range of motion of the fingers was measured. Total active motion was scored based on the American Society for Surgery of the Hand. All measurements were compared with those on the opposite fingers. Patients also reported on their satisfaction using the 100-mm visual analogue scale. RESULTS: At the final follow-ups of 2 years, range of motion of metacarpophalangeal joint, proximal phalangeal joint, and distal interphalangeal joint reached 97%±2.88, 93%±6.65, and 96%±3.22 of the opposite fingers, respectively. Based on Total active motion scoring system, we obtained 36 excellent and 15 good results. Based on VAS, patient satisfaction was 96±3.44. CONCLUSIONS: The cemented K-wire fixation is a reliable technique for the treatment of shaft fractures of middle phalanges. The technique is a minimally invasive procedure with minimal complications. LEVEL OF EVIDENCE: Therapeutic study, Level IVa.

microRNA regulatory mechanism by which PLLA aligned nanofibers influence PC12 cell differentiation.

OBJECTIVE: Aligned nanofibers (AFs) are regarded as promising biomaterials in nerve tissue engineering. However, a full understanding of the biocompatibility of AFs at the molecular level is still challenging. Therefore, the present study focused on identifying the microRNA (miRNA)-mediated regulatory mechanism by which poly-L-lactic acid (PLLA) AFs influence PC12 cell differentiation. APPROACH: Firstly, the effects of PLLA random nanofibers (RFs)/AFs and PLLA films (control) on the biological responses of PC12 cells that are associated with neuronal differentiation were examined. Then, SOLiD sequencing and cDNA microarray were employed to profile the expressions of miRNAs and mRNAs. The target genes of the misregulated miRNAs were predicted and compared with the mRNA profile data. Functions of the matched target genes (the intersection between the predicted target genes and the experimentally-determined, misregulated genes) were analyzed. MAIN RESULTS: The results revealed that neurites spread in various directions in control and RF groups. In the AF group, most neurites extended in parallel with each other. The glucose consumption and lactic acid production in the RF and AF groups were higher than those in the control group. Compared with the control group, 42 and 94 miRNAs were significantly dysregulated in the RF and AF groups, respectively. By comparing the predicted target genes with the mRNA profile data, five and 87 matched target genes were found in the RF and AF groups, respectively. Three of the matched target genes in the AF group were found to be associated with neuronal differentiation, whereas none had this association in the RF group. The PLLA AFs induced the dysregulation of miRNAs that regulate many biological functions, including axonal guidance, lipid metabolism and long-term potentiation. In particular, two miRNA-matched target gene-biological function modules associated with neuronal differentiation were identified as follows: (1) miR-23b, miR-18a, miR-107 and miR-103 regulate the Rras2 and Nf1 gene and thereby, affect cytoskeleton regulation and MAPK pathway; (2) miR-92a, miR-339-5p, miR-25, miR-125a-5p, miR-351 and miR-19b co-regulate the Pafah1b1 gene, affecting PC12 cell migration and differentiation. SIGNIFICANCE: This work demonstrates a bioinformatic approach to accomplish miRNA-mRNA profile integrative analysis and provides more insights for understanding the regulatory mechanism of miRNA in AFs affecting neuronal differentiation. These findings will be greatly beneficial for the application and design of AFs in nerve tissue engineering.

Influence of Poly(L-Lactic Acid) Aligned Nanofibers on PC12 Differentiation.

The aim of this study was to unveil the mechanism by which aligned nanofibers influence neuronal differentiation. PC12 cells were seeded on three different poly(L-lactic acid) (PLLA) substrates (PLLA films (control), electrospun PLLA random nanofibers (RF) and electrospun PLLA aligned nanofibers (AF)). Subsequently, cellular experiments, cDNA microarrays and molecular biological approaches were employed to investigate the impacts of the different PLLA substrates on PC12 cell differentiation. Scanning electron microscope observation revealed that neurite outgrowth in the AF group was parallel to the direction of nanofiber alignment and that the filopodias at the neurite tips spread along the aligned nanofiber axis. Meanwhile, both neurite length and the expression of GAP43 (a neuronal differentiation marker gene) were higher in the AF group than those in the control and RF groups. These results suggested that the PLLA aligned nanofibers enhanced PC12 cell differentiation. cDNA microarray experiment revealed that 876 and 1937 genes had significantly changed expression in the RF and AF groups, respectively. Based on gene ontology analysis, 493 and 1193 differentially expressed genes involved in neuronal differentiation were found in the RF and AF groups, respectively. Pathway analysis showed that the PLLA aligned nanofibers mainly mediated their effects via integrin-mediated pathways. qRT-PCR and western blotting assays further confirmed that gene and protein expression levels in the integrin-mediated FAK-MEK-ERK pathway (e.g., Tln1, Mapk6, phosphorylated-ERK1/2) were enhanced by the PLLA aligned nanofibers. Both PC12 cell differentiation and the expressions of genes and proteins in the integrin-mediated FAK-MEK-ERK pathway were inhibited when integrins were blocked by the pentapeptide GRGDS. In addition, the Pafah1b-1 gene was found to be involved in PLLA aligned nanofibers' promotion of PC12 cell differentiation. Taken together, the results suggested that PLLA aligned nanofibers might cooperate with nerve growth factor (NGF) to induce PC12 cell differentiation by activating the integrin-mediated FAK-MEK-ERK pathway and the Pafah1b1 gene.