Teflon promotes chromosomal recruitment of homolog conjunction proteins during Drosophila male meiosis.

Meiosis in males of higher dipterans is achiasmate. In their spermatocytes, pairing of homologs into bivalent chromosomes does not include synaptonemal complex and crossover formation. While crossovers preserve homolog conjunction until anaphase I during canonical meiosis, an alternative system is used in dipteran males. Mutant screening in Drosophila melanogaster has identified teflon (tef) as being required specifically for alternative homolog conjunction (AHC) of autosomal bivalents. The additional known AHC genes, snm, uno and mnm, are needed for the conjunction of autosomal homologs and of sex chromosomes. Here, we have analyzed the pattern of TEF protein expression. TEF is present in early spermatocytes but cannot be detected on bivalents at the onset of the first meiotic division, in contrast to SNM, UNO and MNM (SUM). TEF binds to polytene chromosomes in larval salivary glands, recruits MNM by direct interaction and thereby, indirectly, also SNM and UNO. However, chromosomal SUM association is not entirely dependent on TEF, and residual autosome conjunction occurs in tef null mutant spermatocytes. The higher tef requirement for autosomal conjunction is likely linked to the quantitative difference in the amount of SUM protein that provides conjunction of autosomes and sex chromosomes, respectively. During normal meiosis, SUM proteins are far more abundant on sex chromosomes compared to autosomes. Beyond promoting SUM recruitment, TEF has a stabilizing effect on SUM proteins. Increased SUM causes excess conjunction and consequential chromosome missegregation during meiosis I after co-overexpression. Similarly, expression of SUM without TEF, and even more potently with TEF, interferes with chromosome segregation during anaphase of mitotic divisions in somatic cells, suggesting that the known AHC proteins are sufficient for establishment of ectopic chromosome conjunction. Overall, our findings suggest that TEF promotes alternative homolog conjunction during male meiosis without being part of the final physical linkage between chromosomes.

The use of polymeric meshes for pelvic organ prolapse: Current concepts, challenges, and future perspectives.

Pelvic organ prolapse (POP) is one of the most common chronic disorders in women, impacting the quality of life of millions of them worldwide. More than 100 surgical procedures have been developed over the decades to treat POP. However, the failure of conservative strategies and the number of patients with recurrence risk have increased the need for further adjuvant treatments. Since their introduction, surgical synthetic meshes have dramatically transformed POP repair showing superior anatomic outcomes in comparison to traditional approaches. Although significant progress has been attained, among the meshes in clinical use, there is no single mesh appropriate for every surgery. Furthermore, due to the risk of complications including acute and chronic infection, mesh shrinkage, and erosion of the tissue, the benefits of the use of meshes have recently been questioned. The aim of this work is to review the evolution of POP surgery, analyzing the current challenges, and detailing the key factors pertinent to the design of new mesh systems. Starting with a description of the pelvic floor anatomy, the article then presents the traditional treatments used in pelvic organ disorders. Next, the development of synthetic meshes is described with an insight into how their function is dependent on both mesh design variables (i.e., material, structure, and functional treatment) and surgical applications. These are then linked to common mesh-related complications, and an indication of current research aiming to address these issues.

MYC competes with MiT/TFE in regulating lysosomal biogenesis and autophagy through an epigenetic rheostat.

Coordinated regulation of the lysosomal and autophagic systems ensures basal catabolism and normal cell physiology, and failure of either system causes disease. Here we describe an epigenetic rheostat orchestrated by c-MYC and histone deacetylases that inhibits lysosomal and autophagic biogenesis by concomitantly repressing the expression of the transcription factors MiT/TFE and FOXH1, and that of lysosomal and autophagy genes. Inhibition of histone deacetylases abates c-MYC binding to the promoters of lysosomal and autophagy genes, granting promoter occupancy to the MiT/TFE members, TFEB and TFE3, and/or the autophagy regulator FOXH1. In pluripotent stem cells and cancer, suppression of lysosomal and autophagic function is directly downstream of c-MYC overexpression and may represent a hallmark of malignant transformation. We propose that, by determining the fate of these catabolic systems, this hierarchical switch regulates the adaptive response of cells to pathological and physiological cues that could be exploited therapeutically.

Immobilization of Ochrobactrum tritici As5 on PTFE thin films for arsenite biofiltration.

Ochrobactrum tritici SCII24T bacteria is an environmental strain with high capacity to resist to arsenic (As) toxicity, which makes it able to grow in the presence of As(III). The inactivation of the two functional arsenite efflux pumps, ArsB and ACR3_1, resulted in the mutant O. tritici As5 exhibiting a high accumulation of arsenite. This work describes a method for the immobilization of the mutant cells O. tritici As5, on a commercial polymeric net after sputtered modified by the deposition of poly(tetrafluoroethylene) (PTFE) thin films, and demonstrates the capacity of immobilized cells to accumulate arsenic from solutions. Six different set of deposition parameters for PTFE thin films were developed and tested in vitro regarding their ability to immobilize the bacterial cells. The surface that exhibited a mild zeta potential value, hydrophobic characteristics, the lowest surface free energy but with a high polar component and the appropriate ratio of chemical reactive groups allowed cells to proliferate and to grow as a biofilm. These immobilized cells maintained their ability to accumulate the surrounding arsenite, making it a great arsenic biofilter to be used in bioremediation processes.

Mass spectrometric imaging of in vivo protein and lipid adsorption on biodegradable vascular replacement systems.

Cardiovascular diseases present amongst the highest mortality risks in Western civilization and are frequently caused by arteriosclerotic vessel failure. Coronary artery and peripheral vessel reconstruction necessitates the use of small diameter systems that are mechanically stress-resistant and biocompatible. Expanded polytetrafluorethylene (ePTFE) is amongst the materials used most frequently for non-degradable and bio-degradable vessel reconstruction procedures, with thermoplastic polyurethanes (TPU) representing a promising substitute. The present study describes and compares the biological adsorption and diffusion occurring with both materials following implantation in rat models. Gel electrophoresis and thin-layer chromatography, combined with mass spectrometry and mass spectrometry imaging, were utilized to identify the adsorbed lipids and proteins. The results were compared with the analytes present in native aorta tissue. It was revealed that both polymers were severely affected by biological adsorption after 10 min in vivo. Proteins associated with cell growth and migration were identified, especially on the luminal graft surface, while lipids were found to be located on both the luminal and abluminal surfaces. Lipid adsorption and cholesterol diffusion were found to be correlated with the polymer modifications identified on degradable thermoplastic urethane graft samples, with the latter revealing extensive cholesterol adsorption. The present study demonstrates an interaction between biological matter and both graft materials, and provides insights into polymer changes, in particular, those observed with thermoplastic urethanes already after 10 min in vivo exposure. ePTFE demonstrated minor polymer modifications, whereas several different polymer signals were observed for TPU, all were co-localized with biological signals.

Surgical outcomes of primary and revision augmentation rhinoplasty using a processed fascia lata.

BACKGROUND: Dorsal augmentation is the most commonly performed procedure in rhinoplasty for Asian patients. Due to the anatomic features of the Asian nose, the use of nonautologous materials to obtain a proper degree of augmentation is inevitable in most cases. Because the use of nonautologous materials possesses a higher risk of complications, surgeons are concerned about selecting suitable materials for the procedure, especially in revision rhinoplasty. Therefore, this study was designed to evaluate the suitability and usefulness of a homologous material, Tutoplast-processed fascia lata (TPFL), in revision augmentation rhinoplasty. METHODS: Retrospective analysis of 104 rhinoplasty patients (primary, 86; revision, 18) who had undergone dorsal augmentation using TPFL was conducted. The comparison of surgical outcomes between primary and revision surgery was made using objective [dorsal height (DH) and radix height (RH), complication rate] and subjective (patient satisfaction) parameters. RESULTS: The degree of augmentation represented by DH and RH was comparable between primary and revision rhinoplasty using TPFL. In comparing the rate of postoperative complications, only minor incidents were noted, in six cases after primary surgery and in one case after revision surgery. Patient satisfaction was measured in both primary and revision augmentation, with a significant difference observed between the two groups (40.57 ± 9.25 versus 31.48 ± 7.59; p < 0.05). CONCLUSION: TPFL is a feasible implant material that delivers suitable augmentation and patient satisfaction with minimal morbidity in both primary and revision rhinoplasty.

The interaction of a histidine-rich protein hpn with the membrane mimics: implications for pathologic roles of Hpn in Helicobacter pylori.

BACKGROUND: Hpn is a small histidine-rich protein in Helicobacter pylori. This protein has been shown to play roles in nickel storage and detoxification and to exhibit cytotoxicity to gastric epithelial cells. Hpn can be secreted outside of the bacterium and forms amyloid-like structures. OBJECTIVE: To study the interactions between Hpn and membrane mimics, which may further our understanding of the pathologic roles of this bacterium. METHODS: Various biochemical and biophysical methods, such as secondary structure determination be CD, calcein release assay with fluorescence spectrometry, and Laurdan and Prodan generalized polarization determination have been used to characterize the interaction between Hpn and membrane mimics. RESULTS: Membrane mimics induced the formation of α-helix in Hpn. The interaction disrupts the integrity of the membrane mimics and leads to the release of inner calcein probe. The experiments involving the Laurdan and Prodan fluorescence indicated that increasing the total protein/lipid ratio leads to a less ordered and more hydrated lipid membrane structure close to the water/lipid interface of lipid bilayers modeling the mitochondrial inner membrane. CONCLUSION: The present data indicated that Hpn may take part in the pathological roles of Helicobacter pylori through membrane interactions.

Immobilization of actively thromboresistant assemblies on sterile blood-contacting surfaces.

Rapid one-step modification of thrombomodulin with alkylamine derivatives such as azide, biotin, and PEG is achieved using an evolved sortase (eSrtA) mutant. The feasibility of a point-of-care scheme is demonstrated herein to site-specifically immobilize azido-thrombomodulin on sterilized commercial ePTFE vascular grafts, which exhibit superior thromboresistance compared with commercial heparin-coated grafts in a primate model of acute graft thrombosis.

Effects of collagen prosthesis cross-linking on long-term tissue regeneration following the repair of an abdominal wall defect.

Collagen prostheses used to repair abdominal wall defects, depending on their pretreatment (noncross-linked vs. cross-linked), besides repair may also achieve tissue regeneration. We assessed the host tissue incorporation of different bioprostheses using a new tool that combines immunofluorescence confocal microscopy with differential interference contrast images, making it possible to distinguish newly formed collagen. Partial hernial defects in the abdominal wall of rabbits were repaired using cross-linked/noncross-linked bioprostheses. Expanded polytetrafluoroethylene (ePTFE) was used as control. After 14/30/90/180 days of implant, specimens were taken for microscopy, immunohistochemistry, and quantitative-reverse transcription-polymerase chain reaction to determine host tissue ingrowth and collagen I/III protein and 1a1/3a1 gene expression. Shrinkage and stress resistance were also examined. At 14 days, cross-linked prostheses had suffered significantly less shrinkage than ePTFE or noncross-linked prostheses. Significantly higher shrinkage was recorded for ePTFE in the longer term. Microscopy revealed encapsulation of ePTFE by neoformed tissue, while the bioprostheses became gradually infiltrated by host tissue. Noncross-linked prosthesis showed better tissue ingrowth, more intense inflammatory reaction and more rapid degradation than the cross-linked prostheses. At 14 days, cross-linked prostheses induced up-regulated collagen 1a1 and 3a1 gene expression, while noncross-linked only showed increased collagen III protein expression at 90 days postimplant. At 6 months, the tensile strengths of cross-linked prostheses were significantly greater compared with ePTFE. Our findings demonstrate that despite the cross-linked collagen prostheses promoting less tissue ingrowth than the noncross-linked meshes, they became gradually replaced by good quality host tissue and were less rapidly degraded, leading to improved stress resistance in the long term.

Peptide derived from Pvfp-1 as bioadhesive on bio-inert surface.

Surface property is one important characteristic of materials, especially for ones that are bio-inert but designed for bio-medical application. In this study, we designed a series of peptides and compared their capacities as bioadhesive to improve the surface bioactivity of bio-inert material. The peptides were designed according to the sequence of Perna viridis foot protein 1 (Pvfp-1), one of the Mfp-1s (mussel foot protein 1) which play key roles in wet adhesion of mussel byssus. And the Teflon (PTFE) was chosen as a model of bio-inert material. With adsorption, adhesion and coating analysis, it was found that peptide C2 (M) (derived from the non-repeating region of Pvfp-1, contains modified DOPA) has superior coating and adhesion abilities especially on the bio-inert surface of PTFE. After coating with peptide C2 (M), the cell adhesion and spreading of osteoblast MC3T3-E1 cells on PTFE were significantly improved compared with those on non-coated surface, and the peptide-coating did not show any cell toxicity. Therefore, peptide C2 (M) is effective for improving the bioactivity of bio-inert PTFE, and could be potentially used as a bioadhesive on other bio-inert materials for biomedical application. Moreover, this study also provided new insights in designing other peptide-based bioadhesive materials.

Application of plasma surface modification techniques to improve hemocompatibility of vascular grafts: A review.

Surface modification using plasma processing can significantly change the chemical and physical characteristics of biomaterial surfaces. When used in combination with additional modification techniques such as direct chemical or biochemical methods, it can produce novel biomaterial surfaces, which are anticoagulant, bioactive, and biomimetic in nature. This article reviews recent advances in improving hemocompatibility of biomaterials by plasma surface modification (PSM). The focus of this review is on PSM of the most commonly used polymers for vascular prostheses such as expanded polytetrafluoroethylene (PTFE), polyethylene terephthalate (Dacron(®) ), and next generation of biomaterials, including polyhedral oligomeric silsesquioxane nanocomposite.

Directed cell attachment by tropoelastin on masked plasma immersion ion implantation treated PTFE.

The ability to generate cell patterns on polymer surfaces is critical for the detailed study of cellular biology, the fabrication of cell-based biosensors, cell separation techniques and for tissue engineering. In this study contact tape masking and steel shadow masks were used to exclude plasma immersion ion implantation (PIII) treatment from defined areas of polytetrafluoroethylene (PTFE) surfaces. This process enabled patterned covalent binding of the cell adhesive protein, tropoelastin, without employing chemical linking molecules. Tropoelastin coating rendered the untreated regions cell adhesive and the PIII-treated area non-adhesive, allowing very fine patterning of cell adhesion to PTFE surfaces. A blocking step, such as with BSA or PEG, was not required to prevent cell binding to the underlying PIII-treated regions as tropoelastin coating alone performed this blocking function. Although tropoelastin coated the entire PTFE surface, the cell binding C-terminus of tropoelastin was markedly less solvent exposed on the PIII-treated, hydrophilic regions. The differential exposure of the C-terminus correlated with the patterned distribution of tropoelastin-mediated cell adhesion. This new methodology specifically enables directed cell behavior on a polymer surface using a simple one-step treatment process, by modulating the adhesive activity of a single extracellular matrix protein.

Nitrogen-rich plasma-polymerized coatings on PET and PTFE surfaces improve endothelial cell attachment and resistance to shear flow.

Low seeding efficiency and poor cell retention under flow-induced shear stress limit the effectiveness of in vitro endothelialization strategies for small-diameter vascular grafts. Primary-amine-rich plasma-polymerized coatings (PPE:N) deposited using low- and atmospheric-pressure plasma discharges on PET and PTFE are evaluated for their ability to improve endothelial cells' kinetics and strength of attachment. PPE:N coatings increase cell adhesion and adhesion rate, spreading, focal adhesion, and resistance to flow-induced shear compared with bare and gelatin-coated PET and PTFE. In particular, about 90% of the cells remain on coated surfaces after 1 h exposure to shear. These coatings, therefore, appear as a promising versatile approach to improve cell seeding strategies for vascular grafts.

Binding of the cell adhesive protein tropoelastin to PTFE through plasma immersion ion implantation treatment.

The interaction of proteins and cells with polymers is critical to their use in scientific and medical applications. In this study, plasma immersion ion implantation (PIII) was used to modify the surface of polytetrafluorethylene (PTFE), enabling the covalent binding of a cell adhesive protein, tropoelastin, without employing chemical linking molecules. Tropoelastin coating of untreated or PIII treated PFTE simultaneously promoted and blocked cell interactions respectively, i.e. PIII treatment of the PTFE surface completely inverses the cell interactive properties of bound tropoelastin. This activity persisted over long term storage of the PIII treated surfaces. The integrin binding C-terminus of tropoelastin was markedly less solvent exposed when bound to PIII treated PTFE than untreated PTFE, accounting for the modulation of cell adhesive activity. This presents a new methodology to specifically modulate cell behavior on a polymer surface using a simple one step treatment process, by adjusting the adhesive activity of a single extracellular matrix protein.

Nano-TiO 2 enriched polymeric powder coatings support human mesenchymal cell attachment and growth.

The objective of this study was to utilize ultrafine powder coating technology to prepare (PPC) that can support human mesenchymal cell attachment and growth. Resins were modified with titanium dioxide and polytetrafluoroethylene (PTFE), and enriched with either SiO(2) or TiO(2) nanoparticles (nSiO(2) or nTiO(2)) to create continuous PPC. Scanning electron microscopy (SEM) revealed complex surface topographies with nano features, and energy dispersive X-ray (EDX) analysis with Ti mapping confirmed a homogenous dispersion of the material. SEM and inverted fluorescence microscopy showed that human embryonic palatal mesenchymal (HEPM) cells attached and spread out on the PPC surfaces, particularly those enriched with nTiO( 2). Cell counts were higher, and the MTT assay measured more metabolic activity from the nTiO(2) enriched PPCs. Furthermore, these cellular responses were enhanced on PPC surfaces that were enriched with a higher concentration of nTiO(2) (2% vs. 0.5%), and appeared comparable to that seen on commercially pure titanium (cpTi). Therefore the nTiO( 2) enrichment of PPC was shown to favor human mesenchymal cell attachment and growth. Indeed, this modification of the materials created continuous surface coatings that sustained a favorable cellular response.

Encapsulation of ePTFE in prevascularized collagen leads to peri-implant vascularization with reduced inflammation.

During the typical healing response to an implanted biomaterial, vascular-rich granulation tissue forms around the implant and later resolves into a relatively avascular, fibrous capsule. We have previously shown that a microvascular construct (MVC) consisting of isolated microvessel fragments suspended in a collagen I gel forms a persistent microcirculation in lieu of avascular scar when implanted. The current study evaluated the potential for microvascular constructs to maintain a vascularized tissue environment around an implanted biomaterial. An analysis of the peri-implant tissue around bare expanded polytetrafluoroethylene (ePTFE), ePTFE embedded within a microvascular construct, or ePTFE embedded within collagen alone revealed that the presence of the MVC, but not collagen alone, promoted vascular densities comparable to that of the granulation tissue formed around bare ePTFE. The vessels within the microvascular construct surrounding the ePTFE were perfusion competent, as determined by India ink perfusion casting, and extended into the interstices of the polymer. In contrast to bare ePTFE, the presence of the MVC or collagen alone significantly reduced the number of activated macrophages in association with ePTFE. Similar results were observed for ePTFE modified to increase cellularity and prevent the formation of an avascular scar. The microvascular construct may prove effective in forming vascularized tissue environments and limiting the number of activated macrophages around implanted polymers thereby leading to effective implant incorporation.

Biomaterial topography alters healing in vivo and monocyte/macrophage activation in vitro.

The effect of biomaterial topography on healing in vivo and monocyte/macrophage stimulation in vitro was assessed. A series of expanded polytetrafluoroethylene (ePTFE) materials were characterized by increasing average intranodal distance of 1.2 µm (1.2-ePTFE), 3.0 µm (3.0-ePTFE), and 4.4 µm (4.4-ePTFE), but presented consistent surface chemistry with nonporous PTFE (np-PTFE). Subcutaneous implantation of 4.4-ePTFE into mice resulted in a statistically thinner capsule that appeared less organized and less dense than the np-PTFE response. In vitro, isolated monocytes/macrophages cultured on np-PTFE produced low levels of interleukin 1-beta (IL-1ß), 1.2-ePTFE and 3.0-ePTFE stimulated intermediate levels, and 4.4-ePTFE stimulated a 15-fold increase over np-PTFE. Analysis of cDNA microarrays demonstrated that additional proinflammatory cytokines and chemokines, including IL-1ß, interleukin 6, tumor necrosis factor alpha, monocyte chemotactic protein 1, and macrophage inflammatory protein 1-beta, were expressed at higher levels by monocytes/macrophages cultured on 4.4-ePTFE at 4 and 24 h, respectively. Expression ratios for several genes were quantified by RT-PCR and were consistent with those from the cDNA array results. These results demonstrate the effect of biomaterial topography on early proinflammatory cytokine production and gene transcription by monocytes/macrophages in vitro and decreased fibrous capsule thickness in vivo.

Friction transfer of polytetrafluoroethylene (PTFE) to produce nanoscale features and influence cellular response in vitro.

A large number of cell types are known to respond to chemical and topographical patterning of substrates. Friction transfer of polytetrafluoroethylene (PTFE) onto substrates has been shown to produce continuous, straight, parallel nanofibres. Ammonia plasma treatment can be used to defluorinate the PTFE, decreasing the dynamic contact angle. Fibroblast and epithelial cells were elongated and oriented with their long axis parallel to the fibres, both individually and in clusters. The fibres restricted cell migration. Cell alignment was slightly reduced on the plasma-treated fibres. These results indicated that although surface topography can affect cellular response, surface chemistry also mediates the extent of this response.

Performance and microbial ecology of air-cathode microbial fuel cells with layered electrode assemblies.

Microbial fuel cells (MFCs) can be built with layered electrode assemblies, where the anode, proton exchange membrane (PEM), and cathode are pressed into a single unit. We studied the performance and microbial community structure of MFCs with layered assemblies, addressing the effect of materials and oxygen crossover on the community structure. Four MFCs with layered assemblies were constructed using Nafion or Ultrex PEMs and a plain carbon cloth electrode or a cathode with an oxygen-resistant polytetrafluoroethylene diffusion layer. The MFC with Nafion PEM and cathode diffusion layer achieved the highest power density, 381 mW/m(2) (20 W/m(3)). The rates of oxygen diffusion from cathode to anode were three times higher in the MFCs with plain cathodes compared to those with diffusion-layer cathodes. Microsensor studies revealed little accumulation of oxygen within the anode cloth. However, the abundance of bacteria known to use oxygen as an electron acceptor, but not known to have exoelectrogenic activity, was greater in MFCs with plain cathodes. The MFCs with diffusion-layer cathodes had high abundance of exoelectrogenic bacteria within the genus Geobacter. This work suggests that cathode materials can significantly influence oxygen crossover and the relative abundance of exoelectrogenic bacteria on the anode, while PEM materials have little influence on anode community structure. Our results show that oxygen crossover can significantly decrease the performance of air-cathode MFCs with layered assemblies, and therefore limiting crossover may be of particular importance for these types of MFCs.

Osteoblast behavior on polytetrafluoroethylene modified by long pulse, high frequency oxygen plasma immersion ion implantation.

Polytetrafluoroethylene (PTFE) is a commonly used medical polymer due to its biological stability and other attractive properties such as high hardness and wear resistance. However, the low surface energy and lack of functional groups to interact with the cellular environment have severely limited its applications in bone or cartilage replacements. Plasma immersion ion implantation (PIII) is a proven effective surface modification technique. However, when conducted on polymeric substrates, conventional PIII experiments typically employ a low pulsing frequency and short pulse duration in order to avoid sample overheating, charging, and plasma sheath extension. In this paper, a long pulse, high frequency O(2) PIII process is described to modify PTFE substrates by implementing a shielded grid in the PIII equipment without these aforementioned adverse effects. X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and contact angle measurements are carried out to reveal the surface effects of PTFE after long pulse, high frequency O(2) PIII and the results are compared to those obtained from conventional short pulse, low frequency O(2) PIII, O(2) plasma immersion, and the untreated control samples. Our results show that less oxygen-containing, rougher, and more hydrophobic surfaces are produced on PTFE after long pulse, high frequency O(2) PIII compared to the other 2 treatments. Cell viability assay, ALP activity test, and real-time PCR analysis are also performed to investigate the osteoblast behavior. It is clear that all 3 surface modification techniques promote osteoblast adhesion and proliferation on the PTFE substrates. Improvements on the ALP, OPN, and ON expression of the seeded osteoblasts are also obvious. However, among these treatments, only long pulse, high frequency O(2) PIII can promote the OCN expression of osteoblasts when the incubation time is 12 days. Our data unequivocally disclose that the long pulse, high frequency O(2) PIII technique is better than the other two types of traditional plasma treatment in the development of PTFE for bone or cartilage repair.