Serous Tubal Intraepithelial Carcinoma Associated With Extraovarian Metastases.

OBJECTIVE: The evolving knowledge of ovarian carcinogenesis sets the stage for our understanding of high-grade serous pelvic carcinoma (HGSC). Findings in prophylactic surgery introduced serous tubal intraepithelial carcinoma (STIC) as potential precursor of HGSC. The present study explores whether STIC instead should already be considered as an early stage of HGSC with a need for comprehensive staging and therapy. PATIENTS AND METHODS: We identified all consecutive patients with HGSC who received first-line therapy in our referral center for gynecologic oncology from January 2011 to April 2016. All chemo-naive patients with upfront debulking surgery in whom an association of STIC and tumor lesions could be analyzed were included. Patients with previous removal of the adnexa or overgrown of the fallopian tube by the tumor were excluded. Pathological workup of the fallopian tubes according to the SEE-FIM protocol was conducted. RESULTS: We analyzed a series of 231 consecutive patients with HGSC of whom 121 (52.4%) had ovarian cancer, 74 (32.0%) had cancer of the fallopian tubes and 36 patients (15.6%) had primary peritoneal cancer. Serous tubal intraepithelial carcinoma could be identified in 158 (68.4%) of 231 patients; of 22 patients, 28.1% is ovarian cancer, 30.8% cancer of the fallopian tubes, and 9.5% peritoneal cancer. Four patients without any further intra-abdominal disease were identified of whom 2 patients had stage FIGO IA and 2 patients had lymph node metastases only. CONCLUSIONS: Our data suggest that STIC should be regarded as a malignant lesion with metastatic potential. Therefore, we recommend a comprehensive surgical staging including lymphadenectomy.

Tuning Surface Defect States in Sputtered Titanium Oxide Electron Transport Layers for Enhanced Stability of Organic Photovoltaics.

Nonfullerene acceptors (NFAs) have dramatically improved the power conversion efficiency (PCE) of organic photovoltaics (OPV) in recent years; however, their device stability currently remains a bottleneck for further technological progress. Photocatalytic decomposition of nonfullerene acceptor molecules at metal oxide electron transport layer (ETL) interfaces has in several recent reports been demonstrated as one of the main degradation mechanisms for these high-performing OPV devices. While some routes for mitigating such degradation effects have been proposed, e.g., through a second layer integrated on the ETL surface, no clear strategy that complies with device scale-up and application requirements has been presented to date. In this work, it is demonstrated that the development of sputtered titanium oxide layers as ETLs in nonfullerene acceptor based OPV can lead to significantly enhanced device lifetimes. This is achieved by tuning the concentration of defect states at the oxide surface, via the reactive sputtering process, to mitigate the photocatalytic decomposition of NFA molecules at the metal oxide interlayers. Reduced defect state formation at the oxide surface is confirmed through X-ray photoelectron spectroscopy (XPS) studies, while the reduced photocatalytic decomposition of nonfullerene acceptor molecules is confirmed via optical spectroscopy investigations. The PBDB-T:ITIC organic solar cells show power conversion efficiencies of around 10% and significantly enhanced photostability. This is achieved through a reactive sputtering process that is fully scalable and industry compatible.

Degradation Adaptability Assessment of Semisolid Powder Molded Mg-Zn-Mn Alloys for Orthopedic Applications.

Semisolid powder molding was used to prepare the medical Mg-6Zn alloy; in order to further improve its degradation adaptability, 0.5 and 1 wt % Mn were added. Then, the effect of the forming temperature (540, 560, 580, and 600 °C) on the in vitro degradation behavior of the prepared Mg-6Zn-xMn (x = 0.5, 1 wt %) was analyzed, and the optimized alloy was obtained. Finally, the biocompatibility and in vivo degradation performance of the optimized and Mn-free alloys were evaluated. Importantly, single-photon emission tomographic imaging (SPECT/CT) was first applied to monitor the in vivo degradation process. The results show that the corrosion mechanism of the Mn-free alloy is microgalvanic corrosion control with corrosive pitting. After adding Mn, the in vitro degradation rate decreases by half (0.17 ± 0.01 mm/year) as the forming temperature increases to 600 °C, and Mg-6Zn-1Mn prepared at 600 °C is the optimized alloy. Mn addition improves the corrosion product film protection and discontinuous secondary phases, and thus, the corrosion mechanism is changed to corrosive pitting control. Additionally, semisolid powder molding is an easy method to prepare alloys with low average pore interconnectivity (<10%), which is helpful for slowing down the degradation rate. The Mn-containing alloy has better biocompatibility, with a cytotoxicity of grade 0-1, due to its lower degradation rate. The in vivo corrosion rate of the Mn-free alloy is 0.19 mm/year after 28 days of implantation, which was precisely detected by SPECT/CT in real-time. The long-term in vivo degradation adaptability of Mn-free and Mn-containing alloys was not correctly presented, which may be due to the unreasonable bone defect model causing implant displacement. However, both of these alloys cause no obvious inflammation and show good healing. In summary, semisolid powder molding is a potentially promising technique to prepare medical Mg alloys, and nuclear imaging is an effective in vivo degradation evaluation method.

Dynamic in vivo monitoring of fracture healing process in response to magnesium implant with multimodal imaging: pilot longitudinal study in a rat external fixation model.

Rodent models are commonly used in pre-clinical research of magnesium (Mg)-based and other types of biomaterials for fracture treatment. Most studies selected unstable fixation methods, and there is a lack of multimodal longitudinal in vivo monitoring of bone healing. The purpose of this study is to develop a rat femoral fracture model stabilized by external fixation with intra-medullary Mg implant, and to investigate the dynamic bone union process with several imaging techniques offering complementing insights into the process. Pure Mg pins were prepared, followed by an in vitro degradation test. Male Sprague-Dawley rats in the experimental group underwent femoral osteotomy stabilized by external fixators with intra-medullary implantation of Mg pins, and the control group underwent external fixation without intra-medullary implants. Post-operative radiograph, micro-CT and B-mode ultrasonography were acquired directly after surgery, and re-examined at week 4, 8 and 12. Bone tissue volume, in vivo implant degradation, histological staining and MRI images were analyzed using ex vivo samples. Both groups achieved fracture union at week 12, and the dynamic healing process was illustrated by in vivo radiograph, micro-CT and ultrasonography. Bilateral whole femur ex vivo analysis further demonstrated increased ratio of bone tissue volume in the surgical femur with Mg implants, and in vivo degradation of Mg pins was slower than in vitro results. Titanium screws rather than intra-medullary Mg pins were the source of artifact in MRI. This pilot study showed the rat fracture model with external fixation and intra-medullary Mg implantation to be an effective method for dynamic in vivo monitoring of the bone healing process. Future application of the animal model may facilitate pre-clinical translational research of biodegradable orthopaedic implant materials for fracture treatment.

Homologous Recombination Deficiency as an Ovarian Cancer Biomarker in a Real-World Cohort: Validation of Decentralized Genomic Profiling.

The diagnostic evaluation of homologous recombination deficiency (HRD) is central to define targeted therapy strategies for patients with ovarian carcinoma. We evaluated HRD in 514 ovarian carcinoma samples by next-generation sequencing of DNA libraries, including BRCA1/BRCA2 and 26,523 single-nucleotide polymorphisms using the standardized Myriad HRD assay, with the predefined cut point of ≥42 for a positive genomic instability score (GIS). All samples were measured in the central Myriad laboratory and in an academic molecular pathology laboratory. A positive GIS was detected in 196 (38.1%) of tumors, whereas 318 (61.9%) were GIS negative. Combining GIS and BRCA mutations, a total of 200 (38.9%) of the 514 tumors were HRD positive. A positive GIS was significantly associated with high-grade serous histology (P < 0.000001), grade 3 tumors (P = 0.001), and patient age <60 years (P = 0.0003). The concordance between both laboratories for the GIS status was 96.9% (P < 0.000001), with a sensitivity of 94.6% and a specificity of 98.4%. Concordance for HRD status was 97.1% (499 of 514 tumors). The percentage of HRD-positive tumors in our real-life cohort was similar to the proportion observed in the recently published PAOLA-1 trial, with high concordance between central and local laboratories. Our results support introduction of the standardized HRD assay in academic molecular pathology laboratories, thus broadening access to personalized oncology strategies for patients with ovarian cancer worldwide.

Processing of Highly Filled Polymer-Metal Feedstocks for Fused Filament Fabrication and the Production of Metallic Implants.

Fused filament fabrication (FFF) is a new procedure for the production of plastic parts, particularly if the parts have a complex geometry and are only needed in a limited quantity, e.g., in specific medical applications. In addition to the production of parts which are purely composed of polymers, fused filament fabrication can be successfully applied for the preparation of green bodies for sintering of metallic implant materials in medical applications. In this case, highly filled polymer-metal feedstocks, which contain a variety of polymeric components, are used. In this study, we focus on various polymer-metal feedstocks, investigate the rheological properties of these materials, and relate them to our results of FFF experiments. Small amplitudes of shear oscillations reveal that the linear range of the polymer-metal feedstocks under investigation is very small, which is caused by elastic and viscous interactions between the metallic particles. These interactions strongly influence or even dominate the flow properties of the feedstock depending on the applied shear stress. The magnitude of the complex viscosity strongly increases with decreasing angular frequency, which indicates the existence of an apparent yield stress. The viscosity increase caused by the high powder loading needed for sintering limits the maximum printing velocity and the minimum layer height. The apparent yield stress hinders the formation of smooth surfaces in the FFF process and slows down the welding of deposited layers. The influence of composition on the processing parameters (suitable temperature range) and part properties (e.g., surface roughness) is discussed on the basis of rheological data.

The Wuerzburg hybridoma library against Drosophila brain.

This review describes the present state of a project to identify and characterize novel nervous system proteins by using monoclonal antibodies (mAbs) against the Drosophila brain. Some 1,000 hybridoma clones were generated by injection of homogenized Drosophila brains or heads into mice and fusion of their spleen cells with myeloma cells. Testing the mAbs secreted by these clones identified a library of about 200 mAbs, which selectively stain specific structures of the Drosophila brain. Using the approach "from antibody to gene", several genes coding for novel proteins of the presynaptic terminal were cloned and characterized. These include the "cysteine string protein" gene (Csp, mAb ab49), the "synapse-associated protein of 47 kDa" gene (Sap47, mAbs nc46 and nb200), and the "Bruchpilot" gene (brp, mAb nc82). By a "candidate" approach, mAb nb33 was shown to recognize the pigment dispersing factor precursor protein. mAbs 3C11 and pok13 were raised against bacterially expressed Drosophila synapsin and calbindin-32, respectively, after the corresponding cDNAs had been isolated from an expression library by using antisera against mammalian proteins. Recently, it was shown that mAb aa2 binds the Drosophila homolog of "epidermal growth factor receptor pathway substrate clone 15" (Eps15). Identification of the targets of mAbs na21, ab52, and nb181 is presently attempted. Here, we review the available information on the function of these proteins and present staining patterns in the Drosophila brain for classes of mAbs that either bind differentially in the eye, in neuropil, in the cell-body layer, or in small subsets of neurons. The prospects of identifying the corresponding antigens by various approaches, including protein purification and mass spectrometry, are discussed.

A novel endotoxin-induced pathway: upregulation of heme oxygenase 1, accumulation of free iron, and free iron-mediated mitochondrial dysfunction.

Mitochondria are involved in the development of organ failure in critical care diseases. However, the mechanisms underlying mitochondrial dysfunction are not clear yet. Inducible hemoxygenase (HO-1), a member of the heat shock protein family, is upregulated in critical care diseases and considered to confer cytoprotection against oxidative stress. However, one of the products of HO-1 is Fe2+ which multiplies the damaging potential of reactive oxygen species catalyzing Fenton reaction. The aim of this study was to clarify the relevance of free iron metabolism to the oxidative damage of the liver in endotoxic shock and its impact on mitochondrial function. Endotoxic shock in rats was induced by injection of lipopolysaccharide (LPS) at a dose of 8 mg/kg (i.v.). We observed that the pro-inflammatory cytokine TNF-alpha and the liver necrosis marker aspartate aminotransferase were increased in blood, confirming inflammatory response to LPS and damage to liver tissue, respectively. The levels of free iron in the liver were significantly increased at 4 and 8 h after onset of endotoxic shock, which did not coincide with the decrease of transferrin iron levels in the blood, but rather with expression of the inducible form of heme oxygenase (HO-1). The proteins important for sequestering free iron (ferritin) and the export of iron out of the cells (ferroportin) were downregulated facilitating the accumulation of free iron in cells. The temporarily increased concentration of free iron in the liver correlated with the temporary impairment of both mitochondrial function and tissue ATP levels. Addition of exogenous iron ions to mitochondria isolated from control animals resulted in an impairment of mitochondrial respiration similar to that observed in endotoxic shock in vivo. Our data suggest that free iron released by HO-1 causes mitochondrial dysfunction in pathological situations accompanied by endotoxic shock.

The Effect of Surface Treatments on the Degradation of Biomedical Mg Alloys-A Review Paper.

This report reviews the effects of chemical, physical, and mechanical surface treatments on the degradation behavior of Mg alloys via their influence on the roughness and surface morphology. Many studies have been focused on technically-used AZ alloys and a few investigations regarding the surface treatment of biodegradable and Al-free Mg alloys, especially under physiological conditions. These treatments tailor the surface roughness, homogenize the morphology, and decrease the degradation rate of the alloys. Conversely, there have also been reports which showed that rough surfaces lead to less pitting and good cell adherence. Besides roughness, there are many other parameters which are much more important than roughness when regarding the degradation behavior of an alloy. These studies, which indicate the relationship between surface treatments, roughness and degradation, require further elaboration, particularly for biomedical Mg alloy applications.

On the Determination of Magnesium Degradation Rates under Physiological Conditions.

The current physiological in vitro tests of Mg degradation follow the procedure stated according to the ASTM standard. This standard, although useful in predicting the initial degradation behavior of an alloy, has its limitations in interpreting the same for longer periods of immersion in cell culture media. This is an important consequence as the alloy's degradation is time dependent. Even if two different alloys show similar corrosion rates in a short term experiment, their degradation characteristics might differ with increased immersion times. Furthermore, studies concerning Mg corrosion extrapolate the corrosion rate from a single time point measurement to the order of a year (mm/y), which might not be appropriate because of time dependent degradation behavior. In this work, the above issues are addressed and a new methodology of performing long-term immersion tests in determining the degradation rates of Mg alloys was put forth. For this purpose, cast and extruded Mg-2Ag and powder pressed and sintered Mg-0.3Ca alloy systems were chosen. DMEM Glutamax +10% FBS (Fetal Bovine Serum) +1% Penicillin streptomycin was used as cell culture medium. The advantages of such a method in predicting the degradation rates in vivo deduced from in vitro experiments are discussed.

Controlled drug release from antibiotic-loaded layered double hydroxide coatings on porous titanium implants in a mouse model.

As an alternative to degradable organic coatings the possibility of using layered double hydroxides (LDHs) to generate implant coatings for controlled drug delivery was evaluated in vivo and in vitro. Coatings prepared from LDH suspensions dissolved slowly and appeared compatible with cultured cells. LDH coatings loaded with an antibiotic resulted in antibacterial effects in vitro. The LDH coating prolonged the drug release period and improved the proliferation of adherent cells in comparison to pure drug coatings. However, during incubation in physiological solutions the LDH coatings became brittle and pieces occasionally detached from the surface. For stress protection porous titanium implants were investigated as a substrate for the coatings. The pores prevented premature detachment of the coatings. To evaluate the coated porous implants in vivo a mouse model was established. To monitor bacterial infection of implants noninvasive in vivo imaging was used to monitor luminescently labeled Pseudomonas aeruginosa. In this model porous implants with antibiotic-loaded LDH coatings could antagonize bacterial infections for over 1 week. The findings provide evidence that delayed drug delivery from LDH coatings could be feasible in combination with structured implant surfaces.

A membrane-anchored Theileria parva cyclophilin with a non-cleaved amino-terminal signal peptide for entry into the endoplasmic reticulum.

Recent studies suggest that peptidyl-prolyl isomerases of the cyclophilin family, that access the secretory pathway, can be involved in the interaction of parasitic protozoa with mammalian host cells. The amino acid sequence of a cDNA encoding a cyclophilin family member of the intracellular protozoan parasite of cattle Theileria parva contains a conserved C-terminal domain that exhibits 70% amino acid identity to cyclophilin proteins from other organisms, and a unique 60 amino acid novel N-terminal extension. Cell-free expression of the cDNA revealed a 26kDa amino translation product, indicating expression of the N-terminal domain. The protein-coding region contains three short introns, less than 100 base pairs in length and Northern blot analysis demonstrates expression of a single 0.9 kb transcript in the piroplasm and schizont stages. The transcript is present in high abundance in the intra-lymphocytic schizont stage. The recombinant protein binds to immobilized cyclosporin A, a finding consistent with peptidyl-prolyl cis-trans isomerase function in vivo. A predicted N-terminal signal peptide was functional for entry into the eukaryotic secretory transport pathway in a cell-free in vitro transcription/translation system. The C-terminal cyclophilin domain was translocated across the membrane of the endoplasmic reticulum and the uncleaved signal peptide functioned as a membrane anchor.

A Porous TiAl6V4 Implant Material for Medical Application.

Increased durability of permanent TiAl6V4 implants still remains a requirement for the patient's well-being. One way to achieve a better bone-material connection is to enable bone "ingrowth" into the implant. Therefore, a new porous TiAl6V4 material was produced via metal injection moulding (MIM). Specimens with four different porosities were produced using gas-atomised spherical TiAl6V4 with different powder particle diameters, namely, "Small" (<45 µm), "Medium" (45-63 µm), "Mix" (90% 125-180 µm + 10% <45 µm), and "Large" (125-180 µm). Tensile tests, compression tests, and resonant ultrasound spectroscopy (RUS) were used to analyse mechanical properties. These tests revealed an increasing Young's modulus with decreasing porosity; that is, "Large" and "Mix" exhibit mechanical properties closer to bone than to bulk material. By applying X-ray tomography (3D volume) and optical metallographic methods (2D volume and dimensions) the pores were dissected. The pore analysis of the "Mix" and "Large" samples showed pore volumes between 29% and 34%, respectively, with pore diameters ranging up to 175 µm and even above 200 µm for "Large." Material cytotoxicity on bone cell lines (SaOs-2 and MG-63) and primary cells (human bone-derived cells, HBDC) was studied by MTT assays and highlighted an increasing viability with higher porosity.

Microstructure and mechanical behavior of metal injection molded Ti-Nb binary alloys as biomedical material.

The application of titanium (Ti) based biomedical materials which are widely used at present, such as commercially pure titanium (CP-Ti) and Ti-6Al-4V, are limited by the mismatch of Young's modulus between the implant and the bones, the high costs of products, and the difficulty of producing complex shapes of materials by conventional methods. Niobium (Nb) is a non-toxic element with strong ß stabilizing effect in Ti alloys, which makes Ti-Nb based alloys attractive for implant application. Metal injection molding (MIM) is a cost-efficient near-net shape process. Thus, it attracts growing interest for the processing of Ti and Ti alloys as biomaterial. In this investigation, metal injection molding was applied to the fabrication of a series of Ti-Nb binary alloys with niobium content ranging from 10wt% to 22wt%, and CP-Ti for comparison. Specimens were characterized by melt extraction, optical microscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), and transmission electron microscopy (TEM). Titanium carbide formation was observed in all the as-sintered Ti-Nb binary alloys but not in the as-sintered CP-Ti. Selected area electron diffraction (SAED) patterns revealed that the carbides are Ti2C. It was found that with increasing niobium content from 0% to 22%, the porosity increased from about 1.6% to 5.8%, and the carbide area fraction increased from 0% to about 1.8% in the as-sintered samples. The effects of niobium content, porosity and titanium carbides on mechanical properties have been discussed. The as-sintered Ti-Nb specimens exhibited an excellent combination of high tensile strength and low Young's modulus, but relatively low ductility.

Transient increase of free iron in rat livers following hemorrhagic-traumatic shock and reperfusion is independent of heme oxygenase 1 upregulation.

Hemorrhagic-traumatic shock (HTS) followed by reperfusion induces heme oxygenase (HO) 1. Free iron (Fe2+) may cause oxidative stress, if not adequately sequestered. We aimed to characterize HO-1-mediated effects on Fe2+ levels in liver and transferrin-bound iron (TFBI) in plasma following HTS, including laparotomy, bleeding, and inadequate and adequate reperfusion. Anesthetized rats showed upregulated HO-1 mRNA at 40 min after HTS, which was followed by increased HO activity at 3 h after shock. Fe2+ levels were transiently increased at 40 min after shock, a time point when HO activity was not affected yet. Levels of plasma TFBI were higher in HTS animals, showing the highest levels at 40 min after shock, and decreased thereafter. In addition, we modulated HO activity 6 h before HTS by administering an inhibitor (zinc-protoporphyrin IX) or an activator (hemin) of HO. At 18 h after HTS in all shock groups, HO activity was increased, the highest being in the hemin-pretreated group. The zinc-protoporphyrin IX-treated HTS animals showed increased HO-1 mRNA and Fe2+ levels in the liver compared with the untreated HTS animals. Transferrin-bound iron levels were affected by pharmacological modulation before shock. All animals undergoing HTS displayed increased TFBI levels after reperfusion; however, in animals pretreated with hemin, TFBI levels increased less. Our data indicate that increase in Fe2+ levels in liver and plasma early after HTS is not mediated by HO-1 upregulation, but possibly reflects an increased mobilization from internal iron stores or increased cell damage. Thus, upregulation of HO activity by hemin does not increase Fe2+ levels following HTS and reperfusion.

Peritoneal inflammation in pigs is associated with early mitochondrial dysfunction in liver and kidney.

The objective of this study was to investigate early effects of peritoneal inflammation on the mitochondrial function in the vital organs, liver and kidney, and their relation to inflammatory and oxidative stress mediators. The study was performed on 14 domestic pigs. Peritoneal inflammation was induced in anesthetized pigs after a midline laparotomy by autologous feces. Fluid resuscitation maintained a MAP above 60 mmHg. Animals were sacrificed 12 h later, and tissue samples were obtained to determine mitochondrial function, mRNA levels of relevant genes [inducible NO synthase (iNOS), inducible HO (HO-1), tumor necrosis factor-alpha (TNF-alpha)], generation of reactive oxygen species (ROS), and HO-1 activity. We found impaired mitochondrial function in both liver and kidney, based on decreased state 3 respiration in the liver and increased states 2 and 4 respiration in the kidney at 12 h. This was accompanied by increased TNF-alpha protein in the blood and up-regulation of TNF-alpha mRNA in the liver. Free iron was elevated in the liver but not in the kidney. In the kidney, mitochondrial ROS production was increased. Nitric oxide levels in blood remained unchanged, corresponding to unchanged levels of iNOS mRNA expression in liver and kidney. Similarly, HO-1 mRNA and heme oxygenase (HO)-activity were unchanged. The inflammatory response in the absence of characteristic septic symptoms was not associated with morphological organ damage at this early time point. Peritoneal inflammation in pigs caused mitochondrial dysfunction in liver and kidney, preceding signs of organ damage. We did not find proof that mitochondrial dysfunction was due to increased levels of either nitric oxide (NO) or products of HO, but it was accompanied by increased levels of oxidative stress markers.