An Optical Fiber-Based Nanomotion Sensor for Rapid Antibiotic and Antifungal Susceptibility Tests.

The emergence of antibiotic and antifungal resistant microorganisms represents nowadays a major public health issue that might push humanity into a post-antibiotic/antifungal era. One of the approaches to avoid such a catastrophe is to advance rapid antibiotic and antifungal susceptibility tests. In this study, we present a compact, optical fiber-based nanomotion sensor to achieve this goal by monitoring the dynamic nanoscale oscillation of a cantilever related to microorganism viability. High detection sensitivity was achieved that was attributed to the flexible two-photon polymerized cantilever with a spring constant of 0.3 N/m. This nanomotion device showed an excellent performance in the susceptibility tests of Escherichia coli and Candida albicans with a fast response in a time frame of minutes. As a proof-of-concept, with the simplicity of use and the potential of parallelization, our innovative sensor is anticipated to be an interesting candidate for future rapid antibiotic and antifungal susceptibility tests and other biomedical applications.

Evaluating ACGME-accredited addiction psychiatry fellowship online content: A critical analysis of addiction psychiatry fellowship program websites in the US.

Introduction: There is an extreme shortage of addiction psychiatrists and a lack of representation of addiction psychiatry (ADP) fellows from racial/ethnic minoritized backgrounds. ADP fellowship websites are integral in engaging potential applicants. It is therefore critical to understand the quality of engagement that trainees are having with ADP fellowship websites. The aim of this study was to investigate the accessibility and content of ADP fellowship program websites in the U.S. Methods: A list of ADP Fellowship programs was obtained from the Accreditation Council for Graduate Medical Education. A critical textual analysis of 42 unique factors within four categories (accessibility, recruitment, education, and health equity) was performed for each ADP fellowship website. Results: Of 51 ADP fellowships, 47 (92.2%) had websites. Information about social media accounts was largely missing from ADP fellowship websites. For recruitment, program description (95.7%) and program director name (76.6%) were most readily available, while interview day (0.00%) and vacation details (10.6%) were least available. For education, a list of rotations (55.3%) and didactics/lectures (40.4%) were most readily available, while post fellowship placement (6.4%), call schedule (4.3%), and responsibility progression (2.1%) were least available. The most prevalent health equity factors were gender-inclusive language (100%) and an absence of stigmatizing addiction language (100%). The least listed were statements of commitment to health equity (0.0%), antiracism training (2.1%), and harm-reduction strategies (4.3%). Conclusions: There are considerable gaps in the amount and types of information provided by ADP fellowship websites. Many existing websites are poorly interfacing with potential leaders in the field. The development of ADP fellowship websites could serve as a low-cost recruitment tool to engage potential addiction specialists. Our findings underscore the need for ADP fellowships to optimize their websites to engage bourgeoning leaders in addiction and optimize access to more comprehensive information.

Perceptions on navigating ACGME-accredited addiction psychiatry fellowship program websites: A thematic analysis across a race- and gender-diverse pool of potential applicants.

Background: There is an alarming shortage of addiction psychiatrists in the United States. To promote interest in addiction psychiatry (ADP), it is essential to maximize resources available through ADP fellowship websites. The aim of this study was to investigate the perceived adequacy and accessibility of content on ADP fellowship websites and discover what further information is considered important among trainees interested in becoming addiction specialists. Methods: Three virtual focus groups were conducted between January and February 2021 among medical students and residents in diverse geographic regions. Participants were asked about the availability of information on ADP fellowship program websites and other material they would like to see available. Focus groups were recorded, with data transcribed and coded using NVivo 11 and Dedoose. A coding scheme was deductively developed based on the core research questions. Results: The majority of participants (N = 27) identified areas of dissatisfaction with the content currently available on ADP websites. The sample was highly representative of racial and ethnic minoritized trainees (n = 12) and genderqueer/non-binary participants (n = 3). Three major themes were identified and durable across all focus groups: lack of emphasis on diversity/health equity, lack of portrayal of everyday life and activities of fellows, and inadequate representation of curricula. Overwhelmingly, participants identified a dedication to health equity (for example, working with minoritized populations) as a key deciding factor in whether to apply to a particular ADP fellowship. Conclusions: ADP fellowship websites are perceived to have considerable variability in the amount and quality of information. Many do not appear to provide the full spectrum of content desired by diverse potential applicants, such as information regarding current fellows and community-centered initiatives. This is concerning, as it suggests ADP fellowships may be interfacing poorly with burgeoning leaders, especially those from race and gender minoritized backgrounds, neglecting potential opportunities to develop future addiction specialists.

A perspective view on the nanomotion detection of living organisms and its features.

The insurgence of newly arising, rapidly developing health threats, such as drug-resistant bacteria and cancers, is one of the most urgent public-health issues of modern times. This menace calls for the development of sensitive and reliable diagnostic tools to monitor the response of single cells to chemical or pharmaceutical stimuli. Recently, it has been demonstrated that all living organisms oscillate at a nanometric scale and that these oscillations stop as soon as the organisms die. These nanometric scale oscillations can be detected by depositing living cells onto a micro-fabricated cantilever and by monitoring its displacements with an atomic force microscope-based electronics. Such devices, named nanomotion sensors, have been employed to determine the resistance profiles of life-threatening bacteria within minutes, to evaluate, among others, the effect of chemicals on yeast, neurons, and cancer cells. The data obtained so far demonstrate the advantages of nanomotion sensing devices in rapidly characterizing microorganism susceptibility to pharmaceutical agents. Here, we review the key aspects of this technique, presenting its major applications. and detailing its working protocols.

Nucleus replacement could get a new chance with annulus closure.

PURPOSE: Disc herniations are usually treated by decompression of the spinal nerves via a partial nucleotomy. As a consequence of reduced disc height (DH), reduced intradiscal pressure (IDP) and increased range of motion (ROM), accelerated degeneration may occur. Nucleus replacement implants are intended to restore those values, but are associated with the risk of extrusion. METHODS: In six fresh frozen lumbar spinal segments (L2-3/L3-4/L4-5/L5-S1, age median 64.5 years (57-72), Pfirrmann grade 2-3), a prolapse was provoked through a box defect (6 × 10 mm) in the annulus. The herniated nucleus material was removed and replaced by a novel collagen-based nucleus implant. An annulus closure device sealed the defect. ROM, neutral zone (NZ) and IDP were measured in the (1) intact and (2) defect state, (3) postoperatively and (4) after cyclic loading (n = 100,000 cycles) applying pure moments (± 7.5 Nm) in flexion-extension, lateral bending and axial rotation. Additionally, the change in DH was determined. Extrusion of implants or nucleus material was evaluated macroscopically. RESULTS: In all specimens, a prolapse could be provoked which decreased DH. Subsequent nucleotomy changed ROM/NZ and IDP considerably. Initial values could be restored by the implantation. Macroscopically, none of the implants nor nucleus material did migrate after cyclic loading. CONCLUSIONS: In this study, a prolapse followed by a nucleotomy resulted in a biomechanical destabilisation. Implantation of the nucleus replacement combined with an annulus closure restored the intact condition without showing signs of extrusion nor migration after cyclic loading. Hence, nucleus replacements could have a new chance in combination with annulus closure devices.

Correction to: Nucleus replacement could get a new chance with annulus closure.

Unfortunately, Fig. 7 and last paragraph of the result section have been incorrectly published. The complete corrected Fig. 7 and last paragraph of the results part (IDP measurements) have been as follows.

Reactive oxygen species and excitation-contraction coupling in the context of cardiac pathology.

Reactive oxygen species (ROS) are highly reactive oxygen-derived chemical compounds that are by-products of aerobic cellular metabolism as well as crucial second messengers in numerous signaling pathways. In excitation-contraction-coupling (ECC), which links electrical signaling and coordinated cardiac contraction, ROS have a severe impact on several key ion handling proteins such as ion channels and transporters, but also on regulating proteins such as protein kinases (e.g. CaMKII, PKA or PKC), thereby pivotally influencing the delicate balance of this finely tuned system. While essential as second messengers, ROS may be deleterious when excessively produced due to a disturbed balance in Na(+) and Ca(2+) handling, resulting in Na(+) and Ca(2+) overload, SR Ca(2+) loss and contractile dysfunction. This may, in the end, result in systolic and diastolic dysfunction and arrhythmias. This review aims to provide an overview of the single targets of ROS in ECC and to outline the role of ROS in major cardiac pathologies, such as heart failure and arrhythmogenesis. This article is part of a Special Issue entitled "Redox Signalling in the Cardiovascular System"

Microbial Sampling Using Interdental Brushes and Paper Points around Teeth and Implants: A Pilot Study for Comparison.

Inflammatory periodontal and peri-implant diseases follow dysbiotic shifts in a susceptible host. A well-established tool for microbial sample collection is the use of paper points. The purpose of this pilot study was to evaluate the use of interdental brushes compared to paper points. Biofilm samples were collected with paper points and later interdental brushes from ten patients. Five patients were represented with a community periodontal index of treatment needs (CPITN) of 0-2 around the teeth and an implant with PPD ≤ 5 mm and no radiological bone loss. The remaining five patients had a CPITN ≥ 3 and one implant with peri-implantitis. Microbial samples were analyzed with quantitative polymerase chain reaction (qPCR) and next-generation sequencing (NGS). The results showed higher amounts of DNA in samples taken by interdental brushes but also higher Ct values. Both methods detected Filifactor alocis, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola in the majority of samples, while Aggregatibacter actinomycetemcomitans was rarely found. A microbial dysbiosis index showed comparable or higher values in sites with no periodontitis/peri-implantitis with interdental brushes. The results of this pilot study indicate that interdental brushes might be a valid technique for microbial sampling and particularly advantageous in the early detection of dysbiotic shifts around teeth and implants. Larger studies with more participants are needed to validate the proposed microbial sampling method with interdental brushes.

Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach.

Introduction: The environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, further elucidation of the antigenic repertoire of B. pseudomallei during human infection for diagnostic and vaccine purposes is required. The adaptation of B. pseudomallei to very different habitats is reflected by a huge genome and a selective transcriptional response to a variety of conditions. We, therefore, hypothesized that exposure of B. pseudomallei to culture conditions mimicking habitats encountered in the human host might unravel novel antigens that are recognized by melioidosis patients. Methods and results: In this study, B. pseudomallei was exposed to various stress and growth conditions, including anaerobiosis, acid stress, oxidative stress, iron starvation and osmotic stress. Immunogenic proteins were identified by probing two-dimensional Western blots of B. pseudomallei intracellular and extracellular protein extracts with sera from melioidosis patients and controls and subsequent MALDI-TOF MS. Among B. pseudomallei specific immunogenic signals, 90 % (55/61) of extracellular immunogenic proteins were identified by acid, osmotic or oxidative stress. A total of 84 % (44/52) of intracellular antigens originated from the stationary growth phase, acidic, oxidative and anaerobic conditions. The majority of the extracellular and intracellular protein antigens were identified in only one of the various stress conditions. Sixty-three immunoreactive proteins and an additional 38 candidates from a literature screening were heterologously expressed and subjected to dot blot analysis using melioidosis sera and controls. Our experiments confirmed melioidosis-specific signals in 58 of our immunoproteome candidates. These include 15 antigens with average signal ratios (melioidosis:controls) greater than 10 and another 26 with average ratios greater than 5, including new promising serodiagnostic candidates with a very high signal-to-noise ratio. Conclusion: Our study shows that a comprehensive B. pseudomallei immunoproteomics approach, using conditions which are likely to be encountered during infection, can identify novel antibody targets previously unrecognized in human melioidosis.

Puf operon sequences and inferred structures of light-harvesting complexes of three closely related Chromatiaceae exhibiting different absorption characteristics.

Whole cells of the purple sulfur bacterium strain 970 exhibit an unusual absorption peak at 963 nm. Its closest relatives, Thiorhodovibrio (Trv.) winogradskyi DSM6702(T) and strain 06511 display a bacteriochlorophyll (BChl) a absorption peak at 867 nm that is characteristic for most light-harvesting complexes 1 (LHC1) of proteobacteria. The puf operons encoding the LHC1 and reaction center proteins were amplified, cloned, and sequenced, and for the Trv. winogradskyi, strains show the common pufBALMC gene arrangement, whereas strain 970 contains a second pufBA copy downstream of pufC. Only pufB(1)A(1) is transcribed, and the corresponding mRNA fragment had an increased stability. Alignments of the deduced protein sequences showed that the LHC1 polypeptides are closely related to those of Thermochromatium (Tch.) tepidum. A deletion between αHis(0) and αTrp(+11), thought to be responsible for the redshifted Q(y) absorption in Tch. tepidum, was also detected in strain 970 and Trv. winogradskyi, whereas αLys(+12) is replaced by histidine only in strain 970. Based on our structural modeling, the side chain of this αHis is predicted to be in close proximity to the BChl a, suggesting that it exerts a modulating effect on the spectral properties of the highly unusual LHC1 complex of strain 970.

Compilation of different data sets of the Late Neolithic wetland site of Pestenacker and of the adjacent valley depositions.

This document contains data sets of the valley depositions of the Loosbach valley and data of the Late Neolithic wetland site of Pestenacker. It consists of raw data and graphical figures of direct push-based electrical conductivity and colour logs and driving core recoveries as well as hand drilling recoveries presented by Köhler et al. [1]. We reviewed unpublished archaeological profiles to determine the incision levels of former stream phases at Pestenacker site. Here, we provide the new, reusable and accessible data set. The data sets and figures of the valley depositions can be used for further analyses, including statistical ones, to improve the methods of the direct-push sensing and to compare it with the sedimentological features recovered from driving core and hand drillings. In addition, the data set is useful for further issues in Pestenacker as well as in the whole central Europe. Especially in the circum-Alpine region, as a comparison with other pile dwellings or stilt houses built from the Neolithic to the Bronce Age.

CaMKII-dependent SR Ca leak contributes to doxorubicin-induced impaired Ca handling in isolated cardiac myocytes.

Doxorubicin (DOX) is one of the most effective chemotherapeutic agents, but cardiotoxicity limits DOX therapy. Although the mechanisms are not entirely understood, reactive oxygen species (ROS) appear to be involved in DOX cardiotoxicity. Ca/calmodulin dependent protein kinase II (CaMKII) can be activated by ROS through oxidation and is known to contribute to myocardial dysfunction through Ca leakage from the sarcoplasmic reticulum (SR). We hypothesized that CaMKII contributes to DOX-induced defects in intracellular Ca ([Ca](i)) handling. Cardiac myocytes were isolated from wild-type (WT) adult rat hearts and from mouse hearts lacking the predominant myocardial CaMKII isoform (CaMKIIδ(-/-), KO) vs. WT. Isolated cardiomyocytes were investigated 30 min after DOX (10 µmol/L) superfusion, using epifluorescence and confocal microscopy. Intracellular ROS-generation ([ROS](i)) and [Ca](i) handling properties were assessed. In a subset of experiments, KN-93 or AIP (each 1 µmol/L) were used to inhibit CaMKII. Melatonin (Mel, 100 µmol/L) served as ROS-scavenger. Western blots were performed to determine the amount of CaMKII phosphorylation and oxidation. DOX increased [ROS](i) and led to significant diastolic [Ca](i) overload in rat myocytes. This was associated with reduced [Ca](i) transients, a 5.8-fold increased diastolic SR Ca leak and diminished SR Ca content. ROS-scavenging partially rescued Ca handling. Western blots revealed increased CaMKII phosphorylation, but not CaMKII oxidation after DOX. Pharmacological CaMKII inhibition attenuated diastolic [Ca](i) overload after DOX superfusion and led to partially restored [Ca](i) transients and SR Ca content, presumably due to reduced Ca spark frequency. In line with this concept, isoform-specific CaMKIIδ-KO attenuated diastolic [Ca](i) overload and Ca spark frequency. DOX exposure induces CaMKII-dependent SR Ca leakage, which partially contributes to impaired cellular [Ca](i) homeostasis. Pharmacological and genetic CaMKII inhibition attenuated but did not completely abolish the effects of DOX on [Ca](i). In light of the clinical relevance of DOX, further investigations seem appropriate to determine if CaMKII inhibition could reduce DOX-induced cardiotoxicity.

Differential requirements for the chemokine receptor CCR7 in T cell activation during Listeria monocytogenes infection.

Effective priming of T cell responses depends on cognate interactions between naive T cells and professional antigen-presenting cells (APCs). This contact is the result of highly coordinated migration processes, in which the chemokine receptor CCR7 and its ligands, CCL19 and CCL21, play a central role. We used the murine Listeria monocytogenes infection model to characterize the role of the CCR7/CCR7 ligand system in the generation of T cell responses during bacterial infection. We demonstrate that efficient priming of naive major histocompatibility complex (MHC) class Ia-restricted CD8+ T cells requires CCR7. In contrast, MHC class Ib-restricted CD8+ T cells and MHC class II-restricted CD4+ T cells seem to be less dependent on CCR7; memory T cell responses are independent of CCR7. Infection experiments with bone marrow chimeras or mice reconstituted with purified T cell populations indicate that CCR7 has to be expressed on CD8+ T cells and professional APCs to promote efficient MHC class Ia-restricted T cell priming. Thus, different T cell subtypes and maturation stages have discrete requirements for CCR7.

Regulatory CD4+CD25+ T cells restrict memory CD8+ T cell responses.

CD4+ T cell help is important for the generation of CD8+ T cell responses. We used depleting anti-CD4 mAb to analyze the role of CD4+ T cells for memory CD8+ T cell responses after secondary infection of mice with the intracellular bacterium Listeria monocytogenes, or after boost immunization by specific peptide or DNA vaccination. Surprisingly, anti-CD4 mAb treatment during secondary CD8+ T cell responses markedly enlarged the population size of antigen-specific CD8+ T cells. After boost immunization with peptide or DNA, this effect was particularly profound, and antigen-specific CD8+ T cell populations were enlarged at least 10-fold. In terms of cytokine production and cytotoxicity, the enlarged CD8+ T cell population consisted of functional effector T cells. In depletion and transfer experiments, the suppressive function could be ascribed to CD4+CD25+ T cells. Our results demonstrate that CD4+ T cells control the CD8+ T cell response in two directions. Initially, they promote the generation of a CD8+ T cell responses and later they restrain the strength of the CD8+ T cell memory response. Down-modulation of CD8+ T cell responses during infection could prevent harmful consequences after eradication of the pathogen.

Identifying proteins bound to native mitotic ESC chromosomes reveals chromatin repressors are important for compaction.

Epigenetic information is transmitted from mother to daughter cells through mitosis. Here, to identify factors that might play a role in conveying epigenetic memory through cell division, we report on the isolation of unfixed, native chromosomes from metaphase-arrested cells using flow cytometry and perform LC-MS/MS to identify chromosome-bound proteins. A quantitative proteomic comparison between metaphase-arrested cell lysates and chromosome-sorted samples reveals a cohort of proteins that were significantly enriched on mitotic ESC chromosomes. These include pluripotency-associated transcription factors, repressive chromatin-modifiers such as PRC2 and DNA methyl-transferases, and proteins governing chromosome architecture. Deletion of PRC2, Dnmt1/3a/3b or Mecp2 in ESCs leads to an increase in the size of individual mitotic chromosomes, consistent with de-condensation. Similar results were obtained by the experimental cleavage of cohesin. Thus, we identify chromosome-bound factors in pluripotent stem cells during mitosis and reveal that PRC2, DNA methylation and Mecp2 are required to maintain chromosome compaction.

Ocular involvement in Adamantiades-Behçet's disease in Berlin, Germany.

BACKGROUND: Epidemiological studies demonstrate a prevalence of Adamantiades-Behçet's disease (ABD) in the range of 0.12-420 per 100,000 inhabitants, with the highest rates in Istanbul, Turkey, and the lowest rates in the USA. Ophthalmological data on the prevalence of ocular involvement are limited for ABD in Germany, because most epidemiological studies are based on rheumatological or dermatological data. Berlin is the city with the highest number of non-native German inhabitants, and its multi-ethnic character renders it uniquely appropriate for epidemiological studies on ABD. METHODS: We retrospectively analyzed all ABD patients seen in our department since 1982. All patients fulfilled the criteria of the International Study Group for Behçet's Disease. We found 140 patients (63 female, 77 male), with a mean follow-up of 6.4 years (0.5-22 years). RESULTS: The mean age was 23 at the first manifestation and 32 when the full-blown disease was noted. The mean age at onset of ocular involvement was 30. Most of the patients were of Turkish (n = 73) or German (n = 34) origin. Fifty-six percent developed ocular involvement, which was the first manifestation in 8.6% and the second manifestation in 19.3% of cases. CONCLUSIONS: More than half the patients developed ocular involvement. The calculated prevalence of ocular involvement in ABD is 1.77/100,000 inhabitants for the municipality of Berlin.

Evaluation of disinfectant efficacy against multidrug-resistant bacteria: A comprehensive analysis of different methods.

BACKGROUND: Multidrug-resistant gram-negative bacteria (MDR-GNB) constitute a threat to health care worldwide. Disinfectants are used to prevent and control the spread of MDR-GNB in a hospital setting but their efficacy might be impaired by bacterial mechanisms that may act on both antimicrobials and disinfectants. Determination of minimum inhibitory concentrations is mainly used to determine bacterial susceptibility against disinfectants, but practical tests on surfaces might be more suitable to predict in-use conditions. Our objective was to compare and evaluate 4 different methods widely used to assess surface disinfectant efficacy. METHODS: The efficacy of benzalkonium chloride (BAC), peracetic acid (PAA), and ethanol (ETH) against multidrug-resistant Acinetobacter, Pseudomonas, and Klebsiella strains was assessed by minimum inhibitory concentration determinations, quantitative suspension tests, qualitative suspension tests, and carrier tests. Test results were compared to ascertain the most appropriate method. RESULTS: ETH, PAA, and BAC were highly effective against MDR-GNB, but we observed marked differences in efficacious concentrations (up to 100-fold) as a function of the test method applied. Minimum inhibitory concentration determination was not reliable for evaluating susceptibility or resistance to BAC. CONCLUSIONS: Surface tests should be used to determine bacterial susceptibility against disinfectants. Moreover, suitable guidelines are needed that allow for the standardization and comparison of bactericidal values obtained by different investigators.

What about informed consent in first-trimester ultrasound screening for Down syndrome?

OBJECTIVES: We evaluate the level of information and informed consent given for the screening of Down syndrome (DS) before the first-trimester ultrasound scan. We evaluate the nature of the medical information given and the patient's understanding. This makes it possible to analyze the existence of the necessary elements (level of information and consent) for the patients to exercise their autonomy as to their decision making during the first-trimester ultrasound scan. METHODS: A total of 350 patients were recruited prospectively in 2 hospitals. Patients were divided into 3 groups according to the results of maternal serum screening for DS. Group 1: Patients at high risk of having a child with DS with the second-trimester maternal serum screening (MSS) (risk >1/250). Group 2: Patients having a low risk (risk <1/250). Group 3: Patients who did not undergo maternal serum screening test. A questionnaire was completed before the medical consultation, to assess the quality of the medical consultation before the ultrasound, the level of prior knowledge and the maternal consent. RESULTS: We excluded 45 of the 350 questionnaires from the analysis because they were incomplete (13.1%). We therefore analyzed a total of 305 questionnaires. The three groups analysed were: Group 1: 89 patients (29.2%) identified as at high risk after MSS. Group 2: 137 patients (44.9%) identified as at low risk accordingly to MSS. Group 3: 79 women (25.9%) who did not undergo maternal serum screening test for diverse reasons. The frequency of genuine informed consent was significantly (p < 0.001) higher in group 3 (56.9%) than in group 1 (52.8%) or group 2 (32.1%). Finally, the number of completely uninformed patients was significantly higher in group 2 (18%) than in the other two groups (7.8% for group 1 and 6.3% for group 3, p < 0.001). With multiple regression analysis, we found that maternal consent depended on 4 independent components: (1) Educational level ( or =10). The negative responses to the various questions calls into question the legitimacy of consent for first-trimester ultrasound scans, particularly as concerns the measurement of nucal translucency.

An Additive Manufacturing Technique for the Facile and Rapid Fabrication of Hydrogel-based Micromachines with Magnetically Responsive Components.

Polyethylene glycol (PEG)-based hydrogels are biocompatible hydrogels that have been approved for use in humans by the FDA. Typical PEG-based hydrogels have simple monolithic architectures and often function as scaffolding materials for tissue engineering applications. More sophisticated structures typically take a long time to fabricate and do not contain moving components. This protocol describes a photolithography method that allows for facile and rapid microfabrication of PEG structures and devices. This strategy involves an in-house developed fabrication stage that allows for the rapid fabrication of 3D structures by building upwards in a layer-by-layer fashion. Independent moving components can also be aligned and assembled onto support structures to form integrated devices. These independent components are doped with superparamagnetic iron oxide nanoparticles that are sensitive to magnetic actuation. In this manner, the fabricated devices can be actuated using external magnets to yield movement of the components within. Hence, this technique allows for the fabrication of sophisticated MEMS-like devices (micromachines) that are composed entirely out of a biocompatible hydrogel, able to function without an onboard power source, and respond to a contact-less method of actuation. This manuscript describes the fabrication of both the fabrication set-up as well as the step-by-step method for the microfabrication of these hydrogels-based MEMS-like devices.

Low Diagnostic Yield of Routine Cerebrospinal Fluid Analysis in Juvenile Stroke.

Background: The diagnostic value of cerebrospinal fluid (CSF) analysis in juvenile stroke, i.e., stroke in young adult patients, is not well studied. We sought to determine the therapeutic impact of routine CSF-analysis in young adults with acute ischemic stroke or transient ischemic attack (TIA). Methods: We abstracted data from patients with acute cerebral ischemia aged 18-45 years who were consecutively admitted to our stroke center between 01/2008 and 12/2015. We routinely performed CSF-analysis in patients with hitherto unknown stroke etiology after complete diagnostic work up. We assessed the frequency and underlying causes of abnormal CSF-findings and their impact on secondary stroke prevention therapy. Results: Among 379 patients (median [IQR:IQR3-IQR1] age 39 [10:43-33] years, 48% female) with acute ischemic stroke (n = 306) or TIA (n = 73), CSF analysis was performed in 201 patients (53%). Of these, 25 patients (12.4 %) had CSF pleocytosis (leucocyte cell count ≥ 5 Mpt/L), that was rated as non-specific (e.g., traumatic lumbar puncture, reactive pleocytosis) in 22 patients. Only 3 patients (1.5% of all patients who underwent CSF-analysis) with CSF-pleocytosis had specific CSF-findings that were related to stroke etiology and affected secondary stroke prevention therapy. Imaging findings had already suggested cerebral vasculitis in two of these patients. Conclusions: The diagnostic yield of routine CSF-analysis in juvenile stroke was remarkably low in our study. Our data suggest that CSF-analysis should only be performed if further findings raise the suspicion of cerebral vasculitis.