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A new direction toward the future of orthopedic implants is to combine biodegradable Mg alloys with permanent Ti to produce selectively biodegradable hybrid joints for advanced tissue engineering. However, the strong galvanic corrosion between Mg and Ti is a major issue to be considered. This work aims to explore plasma electrolytic oxidation (PEO) as a single-step coating treatment to allow for an acceptable degradation behavior of MgTi hybrid systems. To this end, MgTi hybrid joints were produced through the heat treatment of Mg-0.6Ca and commercially pure Ti specimens at 640 °C for 8 h. A single-step PEO treatment was then employed to create a protective layer on the surface of hybrid couples. Even though the scanning electron microscopy (SEM) images showed only a porosity of 6% and 12% within the PEO layers on single Mg and MgTi couples, 3D investigation of the synchrotron-based microtomography data demonstrated a porosity of 18% and 30% with a considerable number of interconnected pores. According to the electrochemical impedance spectroscopy measurements, the impedance modulus at all frequencies on coated MgTi coupled specimens was lower than that on the coated single Mg-0.6Ca and pure Ti. However, the application of PEO treatment significantly decreased the strong galvanic degradation of Mg-0.6Ca in contact with Ti. The results of hydrogen evolution tests revealed that PEO-treated MgTi couples showed a similar degradation behavior as the single alloy during the first day of immersion.

The study is focussing towards Metal Injection Moulding (MIM) of Mg-alloys for biomedical implant applications. Especially the influence of the sintering processing necessary for the consolidation of the finished part is in focus of this study. In doing so, the chosen high strength EZK400 Mg-alloy powder material was sintered using different sintering support bottom plate materials to evaluate the possibility of iron impurity pick up during sintering. It can be shown that iron pick up took place from the steel bottom plate into the specimen. Despite the fact that a separating boron nitrite (BN) barrier layer was used and the Mg-Fe phase diagram is not predicting any significant solubility to each other. As a result of this study a new bottom plate material not harming the sintering and the biodegradation performance of the as sintered material, namely a carbon plate material, was found. Keywords: Magnesium, Mg alloy, Sintering, MIM, Powder metallurgy, Biodegradable implant, EZK400

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.

The mortality rate of newborns with severe congenital heart disease (CHD) has significantly decreased over the past few decades. However, many of these children experience neurological impairments, particularly following a hypoxic cardiac arrest. The use of extracorporeal membrane oxygenation (ECMO) has been considered an effective treatment for severe hypoxia in CHD cases. Various clinical studies have examined the use of ECMO for resuscitation after hypoxic cardiac arrest, but the results have been contradictory, showing a significant incidence of both mortality and morbidity in some studies while others report good outcome. In order to investigate the mechanisms behind brain injury associated with extracorporeal circulation, we have developed a neonatal porcine model of hypoxia-induced cardiac arrest followed by veno-arterial ECMO therapy., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

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- x Mn ( 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.

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., (Copyright © 2022 Association for Molecular Pathology and American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

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.

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.

Some β-Ti alloys, such as Ti-Nb-Ta-Zr (TNTZ) alloys, exhibit a low Young's modulus and excellent biocompatibility. These alloys are promising new generation biomedical implant materials. Selective laser melting (SLM) can further enable customer-specific manufacturing of β-Ti alloys to satisfy the ever-increasing need for enhanced biomedical products. In this study, we quantitatively determined the relationships between porosity, yield strength, and Young's modulus of SLM-prepared TNTZ lattices. The study constitutes a critical step toward understanding the behavior of the lattice and eventually enables tuning the Young's modulus to match that of human bones. Fatigue properties were also investigated on as-printed lattices in terms of the stress limit. The biocompatibility study included a routine evaluation of the relative cell growth rate and a proteomics analysis using a common mouse fibroblast cell line, L929. The results indicated that the as-printed TNTZ samples exhibited evidence of protein proliferation of the L929 cells, particularly P06733, and that those proteins are responsible for biological processes and molecular functions. They in turn may have promoted cell regeneration, cell motility, and protein binding, which at least partially explains the good biocompatibility of the as-printed TNTZ at the protein level. The study highlights the promising applications of additively manufactured TNTZ as a bone-replacing material from mechanical and biocompatibility perspectives., Competing Interests: Declaration of competing interest The authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2020. Published by Elsevier B.V.)

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.

The study is focussing towards Metal Injection Moulding (MIM) of Mg-alloys for biomedical implant applications. Especially the influence of the sintering processing necessary for the consolidation of the finished part is in focus of this study. In doing so, the chosen high strength EZK400 Mg-alloy powder material was sintered using different sintering support bottom plate materials to evaluate the possibility of iron impurity pick up during sintering. It can be shown that iron pick up took place from the steel bottom plate into the specimen. Despite the fact that a separating boron nitrite (BN) barrier layer was used and the Mg-Fe phase diagram is not predicting any significant solubility to each other. As a result of this study a new bottom plate material not harming the sintering and the biodegradation performance of the as sintered material, namely a carbon plate material, was found.

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.

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.

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., (© 2014 Wiley Periodicals, Inc.)

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.

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 β 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., (© 2013 Elsevier Ltd. All rights reserved.)

Despite its non-matching mechanical properties titanium remains the preferred metal implant material in orthopaedics. As a consequence in some cases stress shielding effect occurs, leading to implant loosening, osteopenia, and finally revision surgery. Porous metal scaffolds to allow easier specialised cells ingrowth with mechanical properties closer to the ones of bone can overcome this problem. This should improve healing processes, implant integration, and dynamic strength of implants retaining. Three Ti-6Al-4V materials were metal injection moulded and tailored porosities were effectively achieved. After microstructural and mechanical characterisation, two different primary cells of mesenchymal origin (human umbilical cord perivascular cells and human bone derived cells which revealed to be two pertinent models) as well as one cell line originated from primary osteogenic sarcoma, Saos-2, were bestowed to investigate cell-material interaction on genomic and proteome levels. Biological examinations disclosed that no material has negative impact on early adhesion, proliferation or cell viability. An efficient cell ingrowth into material with an average porosity of 25-50 μm was proved.

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.

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.

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.

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.

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., (Copyright 2004 Elsevier B.V.)

This paper describes the development of a cryo-device for the treatment of tumors. The probes are intended to form an iceball inside of an organ, e.g. the liver, to destroy degenerated cells. After successful preliminary tests and the development and construction of several probes, the emphasis is now being placed on the realization of a complete device prototype which will enable clinical studies to be carried out. Important for the functionality of the device is an integrated temperature sensor inside the probes. The device may also be used for cryo-analgesic purposes in pain treatment.

The 104 kDa microneme-rhoptry protein (p104) is the only known apical complex organelle-specific protein of Theileria parva. p104 exhibits striking structural similarities to circumsporozoite protein and sporozoite surface protein 2 of Plasmodium yoelii. Their primary sequences contain two hydrophobic segments, located at the amino-and the carboxy-terminus. The p104 amino-terminal hydrophobic region was suggested to be a signal peptide for entry into the endoplasmic reticulum and the extreme carboxy-terminal region to function as a membrane anchor. We have studied the biogenesis of p104 in a cell-free expression system and found that p104 is co-translationally transported into membranes derived from endoplasmic reticulum. The amino-terminal signal peptide is not cleaved off and anchors the protein in the membrane with the carboxy-terminal portion translocated into the lumen. We suggest that in vivo p104 is co-translationally integrated into the membrane of the endoplasmic reticulum, from where it is further transported to the microneme-rhoptry complex. Thus, p104 appears to be a suitable marker to study the development of micronemes and rhoptries in T. parva.

The schizont stage of the protozoan parasite Theileria parva induces features characteristic of tumor cells in infected bovine T-cell lines. Most strikingly T. parva-infected cell lines acquire unlimited growth potential in vitro. Their proliferative state is entirely dependent on the presence of a viable parasite within the host cell cytoplasm. It has been postulated that parasite proteins either secreted into the host cell or expressed on the parasite surface membrane are involved in the parasite-host cell interaction. We used an in vitro transcription-translation-membrane translocation system to identify T. parva-derived cDNA clones encoding secretory or membrane proteins. Within 600 clones we found one encoding a 17-kDa protein which is processed by microsomal membranes to a 14-kDa protein (11E), presumably by signal peptidase. The processed form is expressed in the T-cell line TpM803 harboring viable parasites. By immunolocalization we show that the 11E protein mostly resides within the parasite, often in close vicinity to membranous structures, but in addition it appears at the surface membrane. Amino acid sequence comparison suggests that 11E belongs to the glutaredoxin family, but is unique so far in containing a signal sequence for endoplasmic reticulum membrane translocation.

Purpose: To evaluate the differentiation of benign from malignant breast tumors with T2*-weighted perfusion magnetic resonance (MR) imaging (blood volume imaging) versus that with dynamic T1-weighted contrast agent-enhanced MR imaging., Materials and Methods: Ten healthy adult volunteers and 18 adult patients with benign or malignant lesions underwent both conventional T1-weighted dynamic contrast-enhanced breast MR imaging and repetitive first-pass, single-section, dynamic T2*-weighted perfusion MR imaging. Images were obtained before, during, and after injection of 20 mL of gadopentetate dimeglumine; peak gadopentetate dimeglumine concentrations were calculated from the maximal signal intensity loss on T2*-weighted images., Results: No perfusion effect was detectable in healthy breast parenchyma. A strong susceptibility-mediated signal intensity loss occurred in malignant breast tumors. No or only minor perfusion effects were seen in fibroadenomas, in spite of their rapid enhancement at T1-weighted dynamic imaging. Perfusion imaging was possible after conventional dynamic contrast-enhanced breast MR imaging., Conclusion: T2*-weighted perfusion imaging exploits the susceptibility-mediated signal intensity loss of a first-pass bolus of gadopentetate dimeglumine within the capillary bed. First-pass perfusion imaging of breast lesions is feasible. It is promising in the differentiation of benign from malignant, rapidly enhancing lesions.

For cytogenetic investigations short-term cultures of 185 breast carcinomas (135 invasive ductal, 21 invasive lobular, 12 invasive ductal with intraductal components, seven heterogeneous, six intraductal, four invasive ductal and lobular) were prepared. Cytogenetic examinations revealed clonal abnormalities in 39 cases with a predominance of simple numerical chromosome changes, i.e., trisomies of chromosomes 7, 8, and 18. One hundred forty-six tumors did not show clonal abnormalities, but single-cell aberrations other than monosomies occurred in 79 of these tumors. Compared to cells of epithelial hyperplasia of the breast, amniotic fluid cells, and cells from pleomorphic adenomas cultivated using the same medium, the frequency of single-cell trisomies was significantly higher. Trisomy 8 was not only found as a clonal aberration in 10 cases but was also the most frequent non-clonal aberration. Trisomy 7 and 18 were also frequent clonal as well as non-clonal cytogenetic deviations.

A Theileria parva specific full-length cDNA clone, T7, which encodes a protein with more than 60% homology to heat shock protein 90 (hsp90) of other organisms, has been identified. T7 appears to be a single copy gene. The gene is expressed as a protein of 87 kDa in both the sporozoite and schizont stages of T. parva. The protein was not found in the piroplasm stage, although the corresponding transcript was detected, suggesting post-transcriptional regulation of the gene. In the schizont stage the T7 protein is upregulated upon heat shock and localized in the cytoplasm.

The structural proteins of rubella virus consist of a nucleocapsid protein (C) and two membrane-embedded spike glycoproteins (E1 and E2). Since many reports have suggested that rubella virus buds intracellularly, we have examined the intracellular transport of the structural proteins in the absence of virion formation, particularly whether the membrane glycoproteins are retained inside the cell or are transported to the cell surface. We have expressed the structural proteins from cloned cDNA either alone or in different combinations, have examined the intracellular location of the proteins by immunofluorescence and using biochemical methods, and have looked for plasma membrane-localized E1 or E2 using a cell surface biotinylation assay. The C protein was found in the Golgi complex when expressed with E2 and E1; without the membrane glycoproteins, C appeared to remain in the endoplasmic reticulum (ER). When expressed alone, E1 was retained in a pre-Golgi compartment, and was not detected at the cell surface in any cell line. When E2 was expressed alone a small fraction could be detected at the cell surface, but the majority was retained intracellularly, apparently in the ER and the Golgi. Both proteins were transported to the surface when they were expressed together, albeit with low efficiencies in all cell lines. These data suggest that, although neither glycoprotein carries a dominant intracellular retention signal, E2 and E1 are largely retained in the Golgi even when present as a transport-competent heterodimer.