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LDS-MIDs (laser direct structured mechatronic integrated devices) are 3D (three-dimensional) circuit carriers that are used in many applications with a focus on antennas. However, thanks to the rising frequencies of HF (high-frequency) systems in 5G and radar applications up to the mmWave (millimeter wave) region, the requirements regarding the geometrical accuracy and minimal wall thicknesses for proper signal propagation in mmWave circuits became more strict. Additionally, interest in combining those with 3D microstructures like trenches or bumps for optimizing transmission lines and subsequent mounting processes is rising. The change from IM (injection molding) to ICM (injection compression molding) could offer a solution for improving the 3D geometries of LDS-MIDs. To enhance the scientific insight into this process variant, this paper reports on the manufacturing of LDS-MIDs for mmWave applications. Measurements of the warpage, homogeneity of local wall thicknesses, and replication accuracy of different trenches and bumps for mounting purposes are presented. Additionally, the effect of a change in the manufacturing process from IM to ICM regarding the dielectric properties of the used thermoplastics is reported as well as the influence of ICM on the properties of LDS metallization—in particular the metallization roughness and adhesion strength. This paper is then concluded by reporting on the HF performance of CPWs (coplanar waveguides) on LDS-MIDs in comparison to an HF-PCB. [ABSTRACT FROM AUTHOR]

As semi-airborne mineral exploration has a limited budget, it is critical to design experimental procedures that generate data to maximize desired information. We investigated the effects of transmitter–receiver geometries for a variety of anomalies and semi-airborne layouts. Our simulations indicated that flight line spacing of 200 m and a point distance of 100 m provides the optimal trade-off between coverage and survey progress for various targets. Based on the target size and distance between the transmitter and the target, the transmitter length should be at least equal to the length of the target. However, where the distance between the transmitter and the target is more than 1 km, the transmitter length should be at least two or three times the target size. Of similar importance are the location and direction of the transmitter cables, which can have a significant impact on the result of inversion and should be parallel to the target strike. By using more than one transmitter, better results can be obtained. If the strike of the target is known, transmitters should be parallel to each other, and if not, it is better to use perpendicular transmitters. The results of this study showed that the optimal distance between transmitters is 3 km. Our simulations showed that it is even possible to recover targets just below the transmitter in corresponding areas of masked data. [ABSTRACT FROM AUTHOR]

In order to optimize elevated kidney retention of previously reported minigastrin derivatives, we substituted (R)-DOTAGA by DOTA in (R)-DOTAGA-rhCCK-16/-18. CCK-2R-mediated internalization and affinity of the new compounds were determined using AR42J cells. Biodistribution and µSPECT/CT imaging studies at 1 and 24 h p.i. were carried out in AR42J tumor-bearing CB17-SCID mice. Both DOTA-containing minigastrin analogs exhibited 3- to 5-fold better IC50 values than their (R)-DOTAGA-counterparts. natLu-labeled peptides revealed higher CCK-2R affinity than their natGa-labeled analogs. In vivo, tumor uptake at 24 h p.i. of the most affine compound, [19F]F-[177Lu]Lu-DOTA-rhCCK-18, was 1.5- and 13-fold higher compared to its (R)-DOTAGA derivative and the reference compound, [177Lu]Lu-DOTA-PP-F11N, respectively. However, activity levels in the kidneys were elevated as well. At 1 h p.i., tumor and kidney accumulation of [19F]F-[177Lu]Lu-DOTA-rhCCK-18 and [18F]F-[natLu]Lu-DOTA-rhCCK-18 was high. We could demonstrate that the choice of chelators and radiometals has a significant impact on CCK-2R affinity and thus tumor uptake of minigastrin analogs. While elevated kidney retention of [19F]F-[177Lu]Lu-DOTA-rhCCK-18 has to be further addressed with regard to radioligand therapy, its radiohybrid analog, [18F]F-[natLu]Lu-DOTA-rhCCK-18, might be ideal for positron emission tomography (PET) imaging due to its high tumor accumulation at 1 h p.i. and the attractive physical properties of fluorine-18. [ABSTRACT FROM AUTHOR]

In order to enable 18F- and 177Lu-labelling within the same molecule, we introduced a silicon-based fluoride acceptor (SiFA) into the hexa-D-glutamate chain of DOTA-PP-F11N. In addition, minigastrin analogues with a prolonged as well as γ-linked D-glutamate chain were synthesised and evaluated. CCK-2R affinity (IC50, AR42J cells) and lipophilicity (logD7.4) were determined. Biodistribution studies at 24 h post-injection (p.i.) and µSPECT/CT imaging at 1, 4 and 24 h p.i. were carried out in AR42J tumour-bearing CB17-SCID mice. CCK-2R affinity of (R)-DOTAGA-rhCCK-1 to 18 was enhanced with increasing distance between the SiFA building block and the binding motif. Lipophilicity of [177Lu]Lu-(R)-DOTAGA-rhCCK-1 to 18 was higher compared to that of [177Lu]Lu-DOTA-PP-F11N and [177Lu]Lu-CP04. The respective α- and γ-linked rhCCK derivatives revealing the highest CCK-2R affinity were further evaluated in vivo. In comparison with [177Lu]Lu-DOTA-PP-F11N, [177Lu-]Lu-(R)-DOTAGA-rhCCK-9 and -16 exhibited three- to eight-fold increased activity levels in the tumour at 24 h p.i. However, activity levels in the kidneys were elevated as well. We could show that the introduction of a lipophilic SiFA moiety into the hydrophilic backbone of [177Lu]Lu-DOTA-PP-F11N led to a decelerated blood clearance and thus improved tumour retention. However, elevated kidney retention has to be addressed in future studies. [ABSTRACT FROM AUTHOR]

Current radiolabeled gastrin-releasing peptide receptor (GRPR) ligands usually suffer from high accumulation in GRPR-positive organs (pancreas, stomach), limiting tumor-to-background contrast in the abdomen. In novel N4-bombesin derivatives this was addressed by substitutions at the Gln7-Trp8 site within the MJ9 peptide (H-Pip5-phe6-Gln7-Trp8-Ala9-Val10-Gly11-His12-Sta13-Leu14-NH2) either by homoserine (Hse7), β-(3-benzothienyl) alanine (Bta8) or α-methyl tryptophan (α-Me-Trp8), with the aim of optimizing pharmacokinetics. We prepared and characterized the peptide conjugates 6-carboxy-1,4,8,11-tetraazaundecane (N4)-asp-MJ9, N4-asp-[Bta8]MJ9, N4-[Hse7]MJ9 and N4-[α-Me-Trp8]MJ9, and evaluated these compounds in vitro (GRPR affinity via IC50,inverse; internalization; lipophilicity via logD7.4) and in vivo (biodistribution and μSPECT/CT studies at 1 h post injection (p.i.) in PC-3 tumor-bearing CB17-SCID mice). 99mTc-labeling resulted in radiochemical yields (RCYs) > 95%. All 99mTc-labeled MJ9 analogues showed comparable or higher GRPR affinity than the external reference [99mTc]Tc-Demobesin 4. Receptor-bound fractions were noticeably higher than that of the reference. Despite a slightly enhanced lipophilicity, all novel MJ9 derivatives revealed improved in vivo pharmacokinetics compared to the reference. The Bta8-modified ligand revealed the most favorable tumor-to-abdomen contrast at 1 h p.i. Substitutions at the Gln7-Trp8 site within GRPR ligands hold great potential to modify pharmacokinetics for improved imaging. [ABSTRACT FROM AUTHOR]

Here, we longitudinally assessed the ex vivo frequency and phenotype of SARS-CoV-2 membrane protein (aa145–164) epitope-specific CD4+ T-cells of an anti-CD20-treated patient with prolonged viral positivity in direct comparison to an immunocompetent patient through an MHC class II DRB1*11:01 Tetramer analysis. We detected a high and stable SARS-CoV-2 membrane-specific CD4+ T-cell response in both patients, with higher frequencies of virus-specific CD4+ T-cells in the B-cell-depleted patient. However, we found an altered virus-specific CD4+ T-cell memory phenotype in the B-cell-depleted patient that was skewed towards late differentiated memory T-cells, as well as reduced frequencies of SARS-CoV-2-specific CD4+ T-cells with CD45RA− CXCR5+ PD-1+ circulating T follicular helper cell (cTFH) phenotype. Furthermore, we observed a delayed contraction of CD127− virus-specific effector cells. The expression of the co-inhibitory receptors TIGIT and LAG-3 fluctuated on the virus-specific CD4+ T-cells of the patient, but were associated with the inflammation markers IL-6 and CRP. Our findings indicate that, despite B-cell depletion and a lack of B-cell—T-cell interaction, a robust virus-specific CD4+ T-cell response can be primed that helps to control the viral replication, but which is not sufficient to fully abrogate the infection. [ABSTRACT FROM AUTHOR]

Background: The recently emerged SARS-CoV-2 B.1.1.529 lineage and its sublineages (Omicron variant) pose a new challenge to healthcare systems worldwide due to its ability to efficiently spread in immunized populations and its resistance to currently available monoclonal antibody therapies. RT-PCR-based variant tests can be used to screen large sample-sets rapidly and accurately for relevant variants of concern (VOC). The aim of this study was to establish and validate a multiplex assay on the cobas 6800/8800 systems to allow discrimination between the two currently circulating VOCs, Omicron and Delta, in clinical samples. Methods: Primers and probes were evaluated for multiplex compatibility. Analytic performance was assessed using cell culture supernatant of an Omicron variant isolate and a clinical Delta variant sample, normalized to WHO-Standard. Clinical performance of the multiplex assay was benchmarked against NGS results. Results: In silico testing of all oligos showed no interactions with a high risk of primer-dimer formation or amplification of human DNA/RNA. Over 99.9% of all currently available Omicron variant sequences are a perfect match for at least one of the three Omicron targets included in the multiplex. Analytic sensitivity was determined as 19.0 IU/mL (CI95%: 12.9–132.2 IU/mL) for the A67V + del-HV69-70 target, 193.9 IU/mL (CI95%: 144.7–334.7 IU/mL) for the E484A target, 35.5 IU/mL (CI95%: 23.3–158.0 IU/mL) for the N679K + P681H target and 105.0 IU/mL (CI95%: 80.7–129.3 IU/mL) for the P681R target. All sequence variances were correctly detected in the clinical sample set (225/225 Targets). Conclusion: RT-PCR-based variant screening compared to whole genome sequencing is both rapid and reliable in detecting relevant sequence variations in SARS-CoV-2 positive samples to exclude or verify relevant VOCs. This allows short-term decision-making, e.g., for patient treatment or public health measures. [ABSTRACT FROM AUTHOR]

Featured Application: The authors integrated platinum carbon (PtC) nano-tips in a tunneling accelerometer. The use of a focused ion beam (FIB) allows flexible and in situ preparation of the tunneling electrodes and other three-dimensional structures in the nanoscale. To realize quantum tunneling applications with movable electrodes, sharp tips with radii down to several tens of nanometers are necessary. The use of a focused ion beam (FIB) and focused electron beam (FEB) with a gas injection system (GIS) allows the integration of geometries in the nanoscale directly into micro and nano systems. However, the implementation of the tunneling effect clearly depends on the material. In this work, a metal-organic precursor is used. The investigation of the prepared tunneling electrodes enables an insight into FIB/FEB parameters for the realization of quantum tunneling applications. For this purpose, a high-resolution transmission electron microscopy (HRTEM) analysis is performed. The results show a dependence of the material nanostructure regarding platinum (Pt) grain size and distribution in an amorphous carbon matrix from the used beam and the FIB currents. The integration of the tips into a polysilicon (PolySi) beam and measuring the current signal by approaching the tips show significant differences in the results. Moreover, the approach of FEB tips shows a non-contact behavior even when the tips are squeezed together. The contact behavior depends on the grain size, proportion of platinum, and the amount of amorphous carbon in the microstructure, especially at the edge area of the tips. This study shows significant differences in the nanostructure between FIB and FEB tips, particularly for the FIB tips: The higher the ion current, the greater the platinum content, the finer the grain size, and the higher the probability of a tunneling current by approaching the tips. [ABSTRACT FROM AUTHOR]

Packaging represents an important part in the microintegration of sensors based on microelectromechanical system (MEMS). Besides miniaturization and integration density, functionality and reliability in combination with flexibility in packaging design at moderate costs and consequently high-mix, low-volume production are the main requirements for future solutions in packaging. This study investigates possibilities employing printed circuit board (PCB-)based assemblies to provide high flexibility for circuit designs together with film-assisted transfer molding (FAM) to package sensors. The feasibility of FAM in combination with PCB and MEMS as a packaging technology for highly sensitive inertia sensors is being demonstrated. The results prove the technology to be a viable method for damage-free packaging of stress- and pressure-sensitive MEMS. [ABSTRACT FROM AUTHOR]

Background: The recent emergence of distinct and highly successful SARS-CoV-2 lineages has substantial implications for individual patients and public health measures. While next-generation-sequencing is routinely performed for surveillance purposes, RT-qPCR can be used to rapidly rule-in or rule-out relevant variants, e.g., in outbreak scenarios. The objective of this study was to create an adaptable and comprehensive toolset for multiplexed Spike-gene SNP detection, which was applied to screen for SARS-CoV-2 B.1.617 lineage variants. Methods: We created a broad set of single nucleotide polymorphism (SNP)-assays including del-Y144/145, E484K, E484Q, P681H, P681R, L452R, and V1176F based on a highly specific multi-LNA (locked nucleic acid)-probe design to maximize mismatch discrimination. As proof-of-concept, a multiplex-test was compiled and validated (SCOV2-617VOC-UCT) including SNP-detection for L452R, P681R, E484K, and E484Q to provide rapid screening capabilities for the novel B.1.617 lineages. Results: For the multiplex-test (SCOV2-617VOC-UCT), the analytic lower limit of detection was determined as 182 IU/mL for L452R, 144 IU/mL for P681R, and 79 IU/mL for E484Q. A total of 233 clinical samples were tested with the assay, including various on-target and off-target sequences. All SNPs (179/179 positive) were correctly identified as determined by SARS-CoV-2 whole genome sequencing. Conclusion: The recurrence of SNP locations and flexibility of methodology presented in this study allows for rapid adaptation to current and future variants. Furthermore, the ability to multiplex various SNP-assays into screening panels improves speed and efficiency for variant testing. We show 100% concordance with whole genome sequencing for a B.1.617.2 screening assay on the cobas6800 high-throughput system. [ABSTRACT FROM AUTHOR]

Essential quality features of pressure sensors are, among other accuracy-related factors, measurement range, operating temperature, and long-term stability. In this work, these features are optimized for a capacitive pressure sensor with a measurement range of 10 bars. The sensor consists of a metal membrane, which is connected to a PCB and a digital capacitive readout. To optimize the performance, different methods for the joining process are studied. Transient liquid phase bonding (TLP bonding), reactive joining, silver sintering, and electric resistance welding are compared by measurements of the characteristic curves and long-term measurements at maximum pressure. A scanning electron microscope (SEM) with energy-dispersive X-ray spectroscopy (EDX) analysis was used to examine the quality of the joints. The evaluation of the characteristic curves shows the smallest measurement errors for TLP bonding and sintering. For welding and sintering, no statistically significant long-term drift was measured. In terms of equipment costs, reactive joining and sintering are most favorable. With low material costs and short process times, electric resistance welding offers ideal conditions for mass production. [ABSTRACT FROM AUTHOR]

So far, only a few reports about reinfections with SARS-CoV-2 have been published, and they often lack detailed immunological and virological data. We report about a SARS-CoV-2 reinfection with a genetically distinct SARS-CoV-2 variant in an immunocompetent female healthcare worker that has led to a mild disease course. No obvious viral escape mutations were observed in the second virus variant. The infectious virus was shed from the patient during the second infection episode despite the presence of neutralizing antibodies in her blood. Our data indicate that a moderate immune response after the first infection, but not a viral escape, did allow for reinfection and live virus shedding. [ABSTRACT FROM AUTHOR]

CRISPR activation (CRISPRa) has revealed great potential as a tool to modulate the expression of targeted cellular genes. Here, we successfully applied the CRISPRa system to trigger the Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation in latently infected cells by selectively activating ORF50 gene directly from the virus genome. We found that a nuclease-deficient Cas9 (dCas9) fused to a destabilization domain (DD) and 12 copies of the VP16 activation domain (VP192) triggered a more efficient KSHV lytic cycle and virus production when guided to two different sites on the ORF50 promoter, instead of only a single site. To our surprise, the virus reactivation induced by binding of the stable DD-dCas9-VP192 on the ORF50 promoter was even more efficient than reactivation induced by ectopic expression of ORF50. This suggests that recruitment of additional transcriptional activators to the ORF50 promoter, in addition to ORF50 itself, are needed for the efficient virus production. Further, we show that CRISPRa can be applied to selectively express the early lytic gene, ORF57, without disturbing the viral latency. Therefore, CRISPRa-based systems can be utilized to facilitate virus–host interaction studies by controlling the expression of not only cellular but also of specific KSHV genes. [ABSTRACT FROM AUTHOR]

With the observation of spiral organism in the stomach about one hundred years ago a long history of a bacterium started ending up in a worldwide research programs, studies and consensuses. With this bacterium rediscovered by Marshall and Warren and later named Helicobacter pylori, a milestone of research was laid nearly twenty years ago. Helicobacter pylori is now recognized as the main cause of most cases of gastritis and ulcer disease in the stomach and the duodenum. In the course of the Helicobacter pylori research, Helicobacter pylori was found to trigger neoplastic alterations on the ground of the inflammation in the stomach. At first, a large number of publications served to describe the connection between Helicobacter pylori and the low malignant B-cell lymphoma of the mucosa-associated lymphoid tissue (MALT) of the stomach. Furthermore, on the basis of numerous seroepidemiological studies, researchers succeeded in documenting the participation of H. pylori, at least as a co-factor, in the development of gastric carcinoma. From large epidemiological studies made during the last twenty years as well as from microbiological research we have learned much more about this in the beginning nameless and mysterious bacterium.