Multi-material laser powder bed fusion additive manufacturing of concentrated wound stator teeth

Additive manufacturing using Powder Bed Fusion by Laser Beam (PBF-LB) enables products with high design freedom. In addition, the ability to process more than one material in all three spatial directions makes it possible to produce highly functional components in one single process. This article investigates whether multi-material manufacturing using PBF-LB is suitable for producing coils for electric motors, which are designed with integrated cooling channels to increase the power density. For this purpose, the copper alloy CuCr1Zr for the coils and the stainless steel 1.4404 (316L) for the core are processed simultaneously. The component designs were verified using 2D and 3D finite element analysis and then manufactured in a multi-material PBF-LB process. While good electrical conductivity of the copper alloy was achieved by heat treatment, it was found that thermal distortion caused deviations from the nominal geometry. The measurement of the electrical properties showed that this distortion leads to short-circuit currents within the coils and the teeth. On this basis, ideas for solutions were developed, with the help of which the functionality of the coils can be ensured or the power density can also be increased. In addition to adapting the design of the component, this includes processing additional or other materials, such as soft magnetic composites

CXCR2-Blocking Has Context-Sensitive Effects on Rat Glioblastoma Cell Line Outgrowth (S635) in an Organotypic Rat Brain Slice Culture Depending on Microglia-Depletion (PLX5622) and Dexamethasone Treatment

In glioblastoma (GBM), the interplay of different immune cell subtypes, cytokines, and/or drugs shows high context-dependencies. Interrelations between the routinely applied dexamethasone (Dex) and microglia remain elusive. Here, we exploited rat organotypic brain slice co-cultures (OBSC) to examine the effects on a rat GBM cell line (S635) outgrowth resulting from the presence of Dex and pretreatment with the colony-stimulating factor receptor 1 (CSF1-R) inhibitor PLX5622: in native OBSC (without PLX5622-pretreatment), a diminished S635 spheroid outgrowth was observable, whereas Dex-treatment enhanced outgrowth in this condition compared to PLX5622-pretreated OBSC. Screening the supernatants of our model with a proteome profiler, we found that CXCL2 was differentially secreted in a Dex- and PLX5622-dependent fashion. To analyze causal interrelations, we interrupted the CXCL2/CXCR2-axis: in the native OBSC condition, CXCR2-blocking resulted in increased outgrowth, in combination with Dex, we found potentiated outgrowth. No effect was found in the PLX5622-pretreated. Our method allowed us to study the influence of three different factors—dexamethasone, PLX5622, and CXCL2—in a well-controlled, simplified, and straight-forward mechanistic manner, and at the same time in a more realistic ex vivo scenario compared to in vitro studies. In our model, we showed a GBM outgrowth enhancing synergism between CXCR2-blocking and Dex-treatment in the native condition, which was levelled by PLX5622-pretreatment

Phage-mediated Dispersal of Biofilm and Distribution of Bacterial Virulence Genes Is Induced by Quorum Sensing

The microbiome and the phage meta-genome within the human gut are influenced by antibiotic treatments. Identifying a novel mechanism, here we demonstrate that bacteria use the universal communication molecule AI-2 to induce virulence genes and transfer them via phage release. High concentrations (i.e. 100 mu M) of AI-2 promote dispersal of bacteria from already established biofilms, and is associated with release of phages capable of infecting other bacteria. Enterococcus faecalis V583 Delta ABC harbours 7 prophages in its genome, and a mutant deficient in one of these prophages (i.e. prophage 5) showed a greatly reduced dispersal of biofilm. Infection of a probiotic E. faecalis strain without lytic prophages with prophage 5 resulted in increased biofilm formation and also in biofilm dispersal upon induction with AI-2. Infection of the probiotic E. faecalis strain with phage-containing supernatants released through AI-2 from E. faecalis V583 Delta ABC resulted in a strong increase in pathogenicity of this strain. The polylysogenic probiotic strain was also more virulent in a mouse sepsis model and a rat endocarditis model. Both AI-2 and ciprofloxacin lead to phage release, indicating that conditions in the gastrointestinal tract of hospitalized patients treated with antibiotics might lead to distribution of virulence genes to apathogenic enterococci and possibly also to other commensals or even to beneficial probiotic strains

Multi-material laser powder bed fusion additive manufacturing of concentrated wound stator teeth

Additive manufacturing using Powder Bed Fusion by Laser Beam (PBF-LB) enables products with high design freedom. In addition, the ability to process more than one material in all three spatial directions makes it possible to produce highly functional components in one single process. This article investigates whether multi-material manufacturing using PBF-LB is suitable for producing coils for electric motors, which are designed with integrated cooling channels to increase the power density. For this purpose, the copper alloy CuCr1Zr for the coils and the stainless steel 1.4404 (316L) for the core are processed simultaneously. The component designs were verified using 2D and 3D finite element analysis and then manufactured in a multi-material PBF-LB process. While good electrical conductivity of the copper alloy was achieved by heat treatment, it was found that thermal distortion caused deviations from the nominal geometry. The measurement of the electrical properties showed that this distortion leads to short-circuit currents within the coils and the teeth. On this basis, ideas for solutions were developed, with the help of which the functionality of the coils can be ensured or the power density can also be increased. In addition to adapting the design of the component, this includes processing additional or other materials, such as soft magnetic composites

PET imaging of chemokine receptor CXCR4 in patients with primary and recurrent breast carcinoma

Abstract Background CXCR4 is a chemokine receptor frequently overexpressed in invasive breast cancer that has been shown to play a major role in signaling pathways involved in metastasis. The aim of this retrospective analysis was to assess the diagnostic performance of CXCR4-directed PET imaging in patients with breast cancer using the recently introduced CXCR4-targeted PET probe 68Ga-Pentixafor. Results Thirteen patients with first diagnosis of breast cancer, four patients with recurrent disease after primary breast cancer, and one patient with axillary lymph node metastasis of unknown primary underwent CXCR4-targeted PET imaging using 68Ga-Pentixafor. Maximum standardized uptake values (SUVmax) and tumor-to-background (T/B) ratios of tumor lesions were measured and compared with pathological prognostic factors and molecular subtypes. 18F-FDG PET/CT images were available in 8/18 cases and were compared semi-quantitatively. Comparison with CXCR4 expression determined by immunohistochemistry was performed in 7/18 patients. Nine of 13 primary breast cancers were visually detectable on 68Ga-Pentixafor PET images (mean SUVmax of 3.0). The visually undetectable lesions included both cases of invasive lobular carcinoma (ILC) and two cases of invasive carcinoma of no special type (NST) without any hormone receptor and HER2 expression (triple negative). Metastases of recurrent breast cancer and unknown primary cancer were visually detectable in all five cases, exhibiting a mean SUVmax of 3.5. 18F-FDG PET demonstrated higher SUVmax in all patients compared to 68Ga-Pentixafor PET. A correlation between SUVmax obtained from 68Ga-Pentixafor PET and prognostic factors including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) status, proliferation index, tumor grade, or molecular subtypes was not observed. Conclusions CXCR4-directed PET imaging in patients with primary and recurrent breast cancer is feasible; however, tumor detectability is significantly lower compared to 18F-FDG PET. Moreover, we did not find any correlation between aforementioned prognostic factors of breast cancer and CXCR4-targeted tracer accumulation. Based on these results in a small patient cohort, CXCR4-targeted PET imaging does not seem to be suitable as a general diagnostic tool for imaging of breast cancer. Future CXCR4 imaging studies should investigate whether this modality might be useful in more specific applications, e.g., in therapeutic approaches especially under the view of current developments in targeted immune cell and immune checkpoint inhibitory therapy

Isolation of Highly Active Monoclonal Antibodies against Multiresistant Gram-Positive Bacteria

<div><p>Multiresistant nosocomial pathogens often cause life-threatening infections that are sometimes untreatable with currently available antibiotics. Staphylococci and enterococci are the predominant Gram-positive species associated with hospital-acquired infections. These infections often lead to extended hospital stay and excess mortality. In this study, a panel of fully human monoclonal antibodies was isolated from a healthy individual by selection of B-cells producing antibodies with high opsonic killing against <i>E. faecalis</i> 12030. Variable domains (VH and VL) of these immunoglobulin genes were amplified by PCR and cloned into an eukaryotic expression vector containing the constant domains of a human IgG1 molecule and the human lambda constant domain. These constructs were transfected into CHO cells and culture supernatants were collected and tested by opsonophagocytic assay against <i>E. faecalis</i> and <i>S. aureus</i> strains (including MRSA). At concentrations of 600 pg/ml, opsonic killing was between 40% and 70% against all strains tested. Monoclonal antibodies were also evaluated in a mouse sepsis model (using <i>S. aureus</i> LAC and <i>E. faecium</i>), a mouse peritonitis model (using <i>S. aureus</i> Newman and LAC) and a rat endocarditis model (using <i>E. faecalis</i> 12030) and were shown to provide protection in all models at a concentration of 4 μg/kg per animal. Here we present a method to produce fully human IgG1 monoclonal antibodies that are opsonic <i>in vitro</i> and protective <i>in vivo</i> against several multiresistant Gram-positive bacteria. The monoclonal antibodies presented in this study are significantly more effective compared to another monoclonal antibody currently in clinical trials.</p></div

Opsonic killing of sera from healthy human volunteers.

<p>Opsonophagocytic killing against <i>E. faecalis</i> 12030 was assessed in sera at a dilution of 1:100. Statistical analysis was done by ANOVA with Dunnett post test. * p<0.05, ** p<0.01, *** p<0.001. Error bars represent standard error of the mean.</p

Sequences of heavy and light chain of the 4 antibodies isolated after the 4th round of selection.

<p>Sequences have been analyzed with IgBlast.</p><p>Sequences of heavy and light chain of the 4 antibodies isolated after the 4th round of selection.</p

Determination of the target of VH4E and VH8.

<p>(A) Opsonophagocytic Inhibition Assay after absorbing mAbs with <i>E. faecalis</i> 12030 treated with Proteinase K or NaIO₄ (B) Binding of VH4E and VH8 to LTA is shown in an ELISA coating wells with <i>S. aureus</i> LTA (C) Osonophagocytic Inhibition Assay absorbing monoclonal antibodies with LTA from <i>S. aureus</i>. <b>(A)</b> Killing of VH4E and VH8 against <i>E. faecalis</i> 12030 is about 65% (first white or grey column) and killing is completely abolished when bacteria are treated with proteinase K before absorption. Opsonic activity of VH4E and VH8 is not inhibited when mAbs are absorbed with <i>E. faecalis</i> treated with NaIO₄ indicating that a polysaccharide is the target of the mAbs. <b>(B)</b> Binding of VH8 and the mouse anti-lipoteichoic monoclonal antibody to LTA was tested in an ELISA. As coating antigen LTA from <i>S. aureus</i> was used and serum dilutions are indicated in the X axis. Each point represents the average of two measurements. <b>(C)</b> An Opsonophagytosis assay after absorbing monoclonal antibodies with purified LTA confirmed the results of the ELISA. Opsonophagocytic killing was compared to controls from which leukocytes were obtained. Statistical analysis was done by ANOVA with Dunnett post test. * p<0.05, ** p<0.01, *** p<0.001. Error bars represent standard error of the mean.</p