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ObjectivePancreatic ductal adenocarcinoma (PDAC) is characterised by an abundant desmoplastic stroma composed of cancer-associated fibroblasts (CAF) and interspersed immune cells. A non-canonical CD8+T-cell subpopulation producing IL-17A (Tc17) promotes autoimmunity and has been identified in tumours. Here, we evaluated the Tc17 role in PDAC.DesignInfiltration of Tc17 cells in PDAC tissue was correlated with patient overall survival and tumour stage. Wild-type (WT) or Il17ra-/-quiescent pancreatic stellate cells (qPSC) were exposed to conditional media obtained from Tc17 cells (Tc17-CM); moreover, co-culture of Tc17-CM-induced inflammatory (i)CAF (Tc17-iCAF) with tumour cells was performed. IL-17A/F-, IL-17RA-, RAG1-deficient and Foxn1nu/numice were used to study the Tc17 role in subcutaneous and orthotopic PDAC mouse models.ResultsIncreased abundance of Tc17 cells highly correlated with reduced survival and advanced tumour stage in PDAC. Tc17-CM induced iCAF differentiation as assessed by the expression of iCAF-associated genes via synergism of IL-17A and TNF. Accordingly, IL-17RA controlled the responsiveness of qPSC to Tc17-CM. Pancreatic tumour cells co-cultured with Tc17-iCAF displayed enhanced proliferation and increased expression of genes implicated in proliferation, metabolism and protection from apoptosis. Tc17-iCAF accelerated growth of mouse and human tumours in Rag1-/-and Foxn1nu/numice, respectively. Finally, Il17ra-expressed by fibroblasts was required for Tc17-driven tumour growth in vivo.ConclusionsWe identified Tc17 as a novel protumourigenic CD8+T-cell subtype in PDAC, which accelerated tumour growth via IL-17RA-dependent stroma modification. We described a crosstalk between three cell types, Tc17, fibroblasts and tumour cells, promoting PDAC progression, which resulted in poor prognosis for patients.

The IBM Z computing platform is optimized for processing vast amounts of data and transactions with low latency in a highly virtualized and secured environment with sustained processor utilization of over 90%. The platform and its microprocessor chip are designed to deliver consistent system performance, throughput, and response times under all conditions. The innovative cache architecture of the IBM Telum Processor provides low latency, large capacity, and reliable L2 caches. Based on a novel horizontal cache persistence algorithm, these L2 caches also serves as system wide L3 and L4 caches delivering optimal enterprise application performance. When built into an IBM z16 system, these architectural features deliver 11% per-core performance improvement over the prior z15 hardware, running real-world enterprise applications.

The processes leading from disturbed B-cell development to adult B-cell progenitor acute lymphoblastic leukemia (BCP-ALL) remain poorly understood. Here, we describe Irf4−/−mice as prone to developing BCP-ALL with age. Irf4−/−preB-I cells exhibited impaired differentiation but enhanced proliferation in response to IL-7, along with reduced retention in the IL-7 providing bone marrow niche due to decreased CXCL12 responsiveness. Thus selected, preB-I cells acquired Jak3mutations, probably following irregular AID activity, resulting in malignant transformation. We demonstrate heightened IL-7 sensitivity due to Jak3mutants, devise a model to explain it, and describe structural and functional similarities to Jak2mutations often occurring in human Ph-like ALL. Finally, targeting JAK signaling with Ruxolitinib in vivo prolonged survival of mice bearing established Irf4−/−leukemia. Intriguingly, organ infiltration including leukemic meningeosis was selectively reduced without affecting blood blast counts. In this work, we present spontaneous leukemogenesis following IRF4 deficiency with potential implications for high-risk BCP-ALL in adult humans.

The cells and mechanisms involved in blood clot resorption are only partially known. We show that regulatory T cells (Tregs) accumulate in venous blood clots and regulate thrombolysis by controlling the recruitment, differentiation and matrix metalloproteinase (MMP) activity of monocytes. We describe a clot Treg population that forms the matricellular acid– and cysteine-rich protein SPARC (secreted protein acidic and rich in cysteine) and show that SPARC enhances monocyte MMP activity and that SPARC+ Tregs are crucial for blood clot resorption. By comparing different treatment times, we define a therapeutic window of Treg expansion that accelerates clot resorption.

Pre-pregnancy obesity and excessive gestational weight gain (GWG) are risk factors for future maternal and childhood obesity. Maternal obesity is potentially communicated to the fetus in part by the metabolome, altering the child’s metabolic program in early development. Fasting maternal blood samples from 37 singleton pregnancies at 25–28 weeks of gestation were obtained from mothers with pre-pregnancy body mass indexes (BMIs) between 18 and 40 kg/m2. Various health measures including GWG, diet, and physical activity were also assessed. At term (37–42 weeks), a venous umbilical cord sample was obtained. Serum metabolomic profiles were measured using nuclear magnetic resonance spectroscopy as well as a gut and metabolic hormone panel. Maternal and cord serum metabolites were tested for associations with pre-pregnancy BMI, GWG, health outcomes, and gut and metabolic hormones. While cord blood metabolites showed no significant correlation to maternal obesity status or other measured health outcomes, maternal serum metabolites showed distinct profiles for lean, overweight, and obese women. Additionally, four serum metabolites, namely, glutamate, lysine, pyruvate, and valine, allowed prediction of excessive GWG when pre-pregnancy BMI was controlled. Metabolic biomarkers predictive of GWG are reported and, if validated, could aid in the guidance of prenatal weight management plans as the majority of pregnancy weight gain occurs in the third trimester.

Patients who have had a stroke with unknown time of onset have been previously excluded from thrombolysis. We aimed to establish whether intravenous alteplase is safe and effective in such patients when salvageable tissue has been identified with imaging biomarkers.

During the current COVID-19 pandemic, the triage, assessment, and management of patients presenting to the emergency department with critical conditions has become ­challenging. The clinical features of COVID-19 are heterogeneous and subtle in many cases. They may easily be overlooked in the case of other acute diseases. Gastrointestinal symptoms are common in patients with COVID-19 as SARS-CoV-2 is able to enter gastrointestinal epithelial cells. However, these complaints can also be caused by a COVID-19-independent concomitant abdominal pathology. Therefore, patients with acute abdominal pain and fever need to be assessed very thoroughly. Based on a clinical case, we present our approach of managing emergency patients with acute abdomen and concomitant suspicion of ­COVID-19.

Cyclopentadienide derivatives possessing intrinsic helical chirality have only rarely been published in the past. The ligand CpCH, which is well-accessible from inexpensive dibenzosuberenone, is paradigm for such a ligand. Here, the synthesis as well as the spectroscopic and structural characterization of a series of platinum group metal complexes containing this ligand are presented. While alkaline metal salts of CpCH failed in transferring the (CpC)−1unit to this type of metal sites, (CpC)Tl turned out to be an excellent precursor for the synthesis of ruthenium ([(η5-CpC)Ru(η6-arene)]PF6, [(η5-CpC)Ru(NCCH3)3)]PF6), rhodium ((η5-CpC)Rh(η4-COD)), and iridium ((η5-CpC)Ir(η4-COD), [(η5-Cp*)Ir(η5-CpC)]PF6) complexes with CpCligands. EXSY NMR studies were carried out to obtain a deeper insight into the ligand dynamics of CpCcomplexes.

Die Herpes-simplex-Virus-Enzephalitis ist eine schwere neurologische Erkrankung. Nicht immer präsentiert sie sich mit den typischen Kardinalsymptomen Vigilanzminderung in Kombination mit Verwirrtheit und Fieber. Welche Untersuchungen sichern die Diagnose? Welche Therapie sollte noch vor den endgültigen Befunden eingeleitet werden?

The Triggering Receptor Expressed on Myeloid Cells (TREM)-1 is a member of the Immunoglobulin superfamily and an activating receptor mainly expressed on myeloid cells. TREM-1 ligation leads to immediate activation of polymorphonuclear neutrophils (PMN) resulting in degranulation and release of reactive oxygen species as well as various effector molecules. Beyond its role in acute and chronic inflammatory processes, TREM-1 is also involved in cancer emergence and progression probably by alteration of the tumor associated neutrophils (TAN) and macrophages (TAM). Advanced information about the TREM-1 signaling cascade may reveal novel therapeutic approaches, as various kinases can specifically be targeted with kinase inhibitors. Therefore, we investigated the protein tyrosine kinome (PTK) and serine threonine kinome (STK) of human PMN after TREM-1 activation. To gain further insights beyond the signaling cascade, we performed RNAseq to validate the TREM-1 mediated activation and to reveal the TREM-1 mediated transcriptome.

Der 46-jährige Familienvater stellt sich in der Notaufnahme vor. Er leidet seit drei Tagen unter massiven Kopfschmerzen, die auf Ibuprofen oder Paracetamol nur wenig und nur kurz ansprechen. Seit dem Vortag hat er auch Fieber.

Many tumors evolve sophisticated strategies to evade the immune system, and these represent major obstacles for efficient antitumor immune responses. Here we explored a molecular mechanism of metabolic communication deployed by highly glycolytic tumors for immunoevasion. In contrast to colon adenocarcinomas, melanomas showed comparatively high glycolytic activity, which resulted in high acidification of the tumor microenvironment. This tumor acidosis induced Gprotein–coupled receptor–dependent expression of the transcriptional repressor ICER in tumor-associated macrophages that led to their functional polarization toward a non-inflammatory phenotype and promoted tumor growth. Collectively, our findings identify a molecular mechanism of metabolic communication between non-lymphoid tissue and the immune system that was exploited by high-glycolytic-rate tumors for evasion of the immune system.

Intelligent industrial automation devices based on cyber-physical systems can establish ad-hoc networks of manufacturing systems to produce individual products. To realize those, all network-participants, with their offered services and properties need to be described in an explicit semantic. A cloud-based concept is conceived in this paper which includes a semantic to describe the manufacturing systems, their characterization as well as the specified user requirements. A cloud-based concept called “e-Production” maps the abilities of the manufacturing systems with the customer requirements and identifies the best collaborative units for production regarding to energy, costs and other quality characteristics.

The microbiota, the entirety of microorganisms residing in the gut, is increasingly recognized as an environmental factor in the maintenance of health and the development of disease. The objective of this analysis was to model in vivointeractions between gut microbiota and both serum and liver metabolites. Different genotypic models (C57BL/6 and BTBRT+tf/jmice) were studied in combination with significant dietary manipulations (chow vs ketogenic diets) to perturb the gut microbiota. Diet rather than genotype was the primary driver of microbial changes, with the ketogenic diet diminishing total bacterial levels. Fecal but not cecal microbiota profiles were associated with the serum and liver metabolomes. Modeling metabolome–microbiota interactions showed fecal Clostridium leptumto have the greatest impact on host metabolism, significantly correlating with 10 circulating metabolites, including 5 metabolites that did not correlate with any other microbes. C. leptumcorrelated negatively with serum ketones and positively with glucose and glutamine. Interestingly, microbial groups most strongly correlated with host metabolism were those modulating gut barrier function, the primary site of microbe–host interactions. These results show very robust relationships and provide a basis for future work wherein the compositional and functional associations of the microbiome can be modeled in the context of the metabolome.

Am Morgen bemerkt der 58-jährige Abteilungsleiter einer Firma vor dem Spiegel einen hängenden Mundwinkel. Seiner Frau fällt eine verwaschene Sprache auf, und beim Frühstücken rinnt dem Mann der Kaffee aus dem Mund.

Metabolite profiles of individuals possessing either the cardiovascular risk or protective variants of the low-density lipoprotein cholesterol (LDL-C) associated 1p13.3 locus of the SORT1gene (rs646776) were analyzed. Serum metabolites and lipids were assessed using LC–MS-based metabolomics in a healthy young population (n= 138: 95 males, 43 females). Although no significant differences were observed in the combined cohort, divergent sex effects were identified. Females carrying the protective allele showed increased phosphatidylcholines, very long chain fatty acids (>C20), and unsaturated fatty acids. Unsaturated fatty acids are considered to be protective against cardiovascular disease. In contrast, males carrying the protective allele exhibited decreased long-chain fatty acids (≤C20) and sphingomyelins, which is similarly considered to decrease cardiovascular disease risk. No significant changes in clinically assessed lipids such as LDL-C, high-density lipoprotein (HDL-C), total cholesterol, or triglycerides were observed in females, whereas only LDL-C was significantly changed in males. This indicates that, apart from reducing LDL-C, other mechanisms may contribute to the protective effect of the SORT1locus. Thus, the analysis of metabolic biomarkers might reveal early disease development that may be overlooked by relying on standard clinical parameters.

Obgleich die Neurolues eine seit vielen Jahrhunderten bekannte Erkrankung ist, bereitet sie im klinischen Alltag aufgrund ihrer unterschiedlichen klinischen Erscheinungsbilder oft Schwierigkeiten: Neben einer subakuten Manifestation als Meningitis kann es nach Monaten bis Jahren zu einer Meningovaskulitis oder einer progressiven Paralyse mit subakuten, fluktuierenden, psychiatrischen Auffälligkeiten kommen. Zudem kann sich die Neurolues in einem sehr späten Stadium als klassische Tabes dorsalis noch Jahrzehnte nach Infektion manifestieren. Da die Syphilis und gleichzeitig auch die Neurolues in den letzten fünf Jahren in Deutschland wieder an Häufigkeit zugenommen haben, muss sie bei entsprechender Klinik unbedingt wieder vermehrt in die Liste der Differenzialdiagnosen aufgenommen werden.

During the adipogenic differentiation process of mesenchymal stem cells, lipid droplets (LDs) grow slowly by transferring lipids between each other. Recent findings hint at the possibility that a fusion pore is involved. In this study, we analyze lipid transfer data obtained in long-term label-free microscopy studies in the framework of a Hagen-Poiseuille model. The data obtained show a LD fusion process in which the lipid transfer directionality depends on the size difference between LDs, whereas the respective rates depend on the size difference and additionally on the diameter of the smaller LDs. For the data analysis, the viscosity of the transferred material has to be known. We demonstrate that a viscosity-dependent molecular rotor dye can be used to measure LD viscosities in live cells. On this basis, we calculate the diameter of a putative lipid transfer channel which appears to have a direct dependence on the diameter of the smaller of the two participating LDs.

The objective of this study was to investigate the relationship between the concentrations of 19 amino acids, glucose, and seven carboxylic acids in the blood and milk of dairy cows and their correlations with established markers of ketosis. To that end, blood plasma and milk specimens were collected throughout lactation in two breeds of dairy cows of different milk yield. Plasma concentrations of glucose, pyruvate, lactate, α-aminobutyrate, β-hydroxybutyrate (BHBA), and most amino acids, except for glutamate and aspartate, were on average 9.9-fold higher than their respective milk levels. In contrast, glutamate, aspartate, and the Krebs cycle intermediates succinate, fumarate, malate, and citrate were on average 9.1-fold higher in milk than in plasma. For most metabolites, with the exception of BHBA and threonine, no significant correlations were observed between their levels in plasma and milk. Additionally, milk levels of acetone showed significant direct relationships with the glycine-to-alanine ratio and the BHBA concentration in plasma. The marked decline in plasma concentrations of glucose, pyruvate, lactate, and alanine in cows with plasma BHBA levels above the diagnostic cutoff point for subclinical ketosis suggests that these animals fail to meet their glucose demand and, as a consequence, rely increasingly on ketone bodies as a source of energy. The concomitant increase in plasma glycine may reflect not only the excessive depletion of protein reserves but also a potential deficiency of vitamin B6.

The origin and function of CD20+T cells are poorly understood. Here, we characterized CD20+T cells in mice and humans and investigated how they are affected by anti-CD20 antibody treatment. We report that murine CD20+T cells are unable to endogenously express the B cell lineage marker CD20; the development of CD20+T cells in rodents requires the presence of CD20-expressing B cells. Our results demonstrated that both murine and human T cells acquire CD20 from B cells via trogocytosis while being activated by an antigen-presenting B cell. In patients with multiple sclerosis (MS) and mice with experimental autoimmune encephalomyelitis (EAE), expression of CD20 on T cells is associated with an up-regulation of activation markers, proinflammatory cytokines, and adhesion molecules, suggesting high pathogenic potential. Supporting this hypothesis, CD20+T cells expand during active EAE in rodents; furthermore, adoptive transfer of CD20+T cells into EAE-diseased mice worsened histological and clinical severity. Of direct therapeutic relevance, we demonstrate that the exclusive therapeutic elimination of CD20+T cells effectively ameliorates EAE, independent of B cells. The results support the hypothesis that CD20+T cells arise upon B cell–T cell interaction and that depletion of CD20+T cells might contribute to the success of anti-CD20 antibody therapies in MS and other inflammatory disorders.

ABSTRACTDespite antibiotic therapy, acute and long-term complications are still frequent in pneumococcal meningitis. One important trigger of these complications is oxidative stress, and adjunctive antioxidant treatment with N-acetyl-l-cysteine was suggested to be protective in experimental pneumococcal meningitis. However, studies of effects on neurological long-term sequelae are limited. Here, we investigated the impact of adjunctive N-acetyl-l-cysteine on long-term neurological deficits in a mouse model of meningitis. C57BL/6 mice were intracisternally infected with Streptococcus pneumoniae. Eighteen hours after infection, mice were treated with a combination of ceftriaxone and placebo or ceftriaxone and N-acetyl-l-cysteine, respectively. Two weeks after infection, neurologic deficits were assessed using a clinical score, an open field test (explorative activity), a t-maze test (memory function), and auditory brain stem responses (hearing loss). Furthermore, cochlear histomorphological correlates of hearing loss were assessed. Adjunctive N-acetyl-l-cysteine reduced hearing loss after pneumococcal meningitis, but the effect was minor. There was no significant benefit of adjunctive N-acetyl-l-cysteine treatment in regard to other long-term complications of pneumococcal meningitis. Cochlear morphological correlates of meningitis-associated hearing loss were not reduced by adjunctive N-acetyl-l-cysteine. In conclusion, adjunctive therapy with N-acetyl-l-cysteine at a dosage of 300 mg/kg of body weight intraperitoneally for 4 days reduced hearing loss but not other neurologic deficits after pneumococcal meningitis in mice. These results make a clinical therapeutic benefit of N-acetyl-l-cysteine in the treatment of patients with pneumococcal meningitis questionable.

Solution nuclear magnetic resonance (NMR) spectroscopy is widely used to analyze complex mixtures of organic compounds such as biological fluids and tissue extracts. Targeted profiling approaches with reliable compound quantitifcation are hampered, however, by signal overlap and other interferences. Here, we present a tool named MetaboQuant for automated compound quantification from pre-processed 1D and 2D heteronuclear single quantum coherence (HSQC) NMR spectral data and concomitant validation of results. Performance of MetaboQuant was tested on a urinary spike-in data set and compared with other quantification strategies. The use of individual calibration factors in combination with the validation algorithms of MetaboQuant raises the reliability of the quantification results. MetaboQuant can be downloaded at http://genomics.uni-regensburg.de/site/institute/software/metaboquant/as stand-alone software for Windows or run on other operating systems from within Matlab. Separate software for peak fitting and integration is necessary in order to use MetaboQuant.

In the ITRS roadmap [1] increasingly long mask write and cycle time is explicitly addressed as a difficult challenge in mask fabrication for the 16nm technology node and beyond. Write time reduction demands have to be seen in relation to corresponding performance parameters like Line Width Roughness (LWR), resolution, placement as well as CD Uniformity. The previously presented Multi Shaped Beam (MSB) approach [2, 3] is considered a potential solution for high throughput mask write application. In order to fully adapt the MSB concept to future industry's requirements specific optimizations are planned. The key element for achieving write time reduction is a higher probe current at the target, which can be obtained by increasing the number of beamlets as well as applying a higher current density. In the present paper the approach of a 256 beamlet MSB design will be discussed. For a given image field size along with a beamlet number increase both beamlet pitch and size have to be optimized. Out of previous investigations, one finding was that by changing the demagnification after the beam forming section of the MSB column the overall performance can be optimized. Based on first electron-optical simulations for a new final lens a larger demagnification turned out to be advantageous. Stochastic beam blur simulation results for the MSB reduction optics will be presented. During the exposure of a pattern layout the number of used beams, their shape and their distribution within the image field varies, which can lead to space charge distortion effects. In regard to this MSB simulation results obtained for an image field of approximately 10x10ìm² will be presented. For the 256 beamlet MSB design and resist sensitivities of 20C/cm2, 40C/cm2and 100C/cm2write time and LWR simulations have been performed. For MSB pattern data fracturing an optimized algorithm has been used, which increased the beamlet utilization factor (indicates the mean number of beamlets which are used per multi-shot). Finally an update with regard to the required changes of the data path architecture for the 256 beamlet MSB approach will be given. Data integrity as an important aspect of the production worthiness of such a systems will be discussed specifically.

To tune their optical properties, a large variety of pyrrolopyrrole cyanines (PPCys) were synthesized with substitutedheteroaromatics such as quinoline, benzothiazole, and oxazole derivatives as terminal groups. Thus, a broad range of stable, highly fluorescing near‐infrared (NIR) dyes with high absorptivities between 690 to 845 nm is accessible. The large number of newly synthesized compounds allows a detailed discussion of the correlation between molecular structure and the optical properties of the first electronic transition.

The genomic analysis of Streptococcus pneumoniae predicted six putative glutamine uptake systems, which are expressed under in vitro conditions, as shown here by reverse transcription-PCR. Four of these operons consist of glnHPQ, while two lack glnH, which encodes a soluble glutamine-binding protein. Here, we studied the impact of two of these glutamine ATP-binding cassette transporters on S. pneumoniae D39 virulence and phagocytosis, which consist of GlnQ and a translationally fused protein of GlnH and GlnP. Mice infected intranasally with D39gln0411/0412 showed significantly increased survival times and a significant delay in the development of pneumococcal pneumonia compared to those infected with D39, as observed in real time using bioluminescent pneumococci. In a mouse sepsis model, the mutant D39gln0411/0412 showed only moderate but significant attenuation. In contrast, the D39gln1098/1099 knockout strain was massively attenuated in the pneumonia and septicemia mouse infection model. To cause pneumonia or sepsis with D39gln1098/1099, infection doses 100- to 10,000-fold higher than those used for wild-type strain D39 were required. In an experimental mouse meningitis model, D39gln1098/1099 produced decreased levels of white blood cells in cerebrospinal fluid and showed decreased numbers of bacteria in the bloodstream compared to D39 and D39gln0411/0412. Phagocytosis experiments revealed significantly decreased intracellular survival rates of mutants D39gln1098/1099 and D39gln0411/0412 compared to wild-type D39, suggesting that the deficiency of Gln uptake systems impairs resistance to oxidative stress. Taken together, our results demonstrate that both glutamine uptake systems are required for full virulence of pneumococci but exhibit different impacts on the pathogenesis of pneumococci under in vivo conditions.

Currently, dexamethasone is the only adjuvant of proven benefit in bacterial meningitis. Dexamethasone halves the risk of poor outcome, but only in selected patient groups. New therapies based upon an understanding of the pathophysiology are needed. This article summarizes our knowledge on the pathophysiology of bacterial meningitis with special emphasis on pneumococcal meningitis, the experimentally best characterized subtype.

Observations of the cosmic microwave background indicate that baryons account for 5 per cent of the Universe’s total energy content. In the local Universe, the census of all observed baryons falls short of this estimate by a factor of two. Cosmological simulations indicate that the missing baryons have not condensed into virialized haloes, but reside throughout the filaments of the cosmic web (where matter density is larger than average) as a low-density plasma at temperatures of 105−107kelvin, known as the warm–hot intergalactic medium. There have been previous claims of the detection of warm–hot baryons along the line of sight to distant blazars and of hot gas between interacting clusters. These observations were, however, unable to trace the large-scale filamentary structure, or to estimate the total amount of warm–hot baryons in a representative volume of the Universe. Here we report X-ray observations of filamentary structures of gas at 107kelvin associated with the galaxy cluster Abell 2744. Previous observations of this cluster were unable to resolve and remove coincidental X-ray point sources. After subtracting these, we find hot gas structures that are coherent over scales of 8 megaparsecs. The filaments coincide with over-densities of galaxies and dark matter, with 5–10 per cent of their mass in baryonic gas. This gas has been heated up by the cluster’s gravitational pull and is now feeding its core. Our findings strengthen evidence for a picture of the Universe in which a large fraction of the missing baryons reside in the filaments of the cosmic web.

Until recently, IgE-activated mast cells have been regarded merely as effector cells of adaptive immune responses, involved in allergic reactions and mucosal immunity to parasites. Herein, we report that murine dermal mast cells, activated by local administration of a cream containing the synthetic TLR7 ligand imiquimod, are essential to initiate an early inflammatory reaction. The mast-cell–derived cytokines TNF-α and IL-1β play an important role in this process. Furthermore, TLR7-activated mast cells are also able to promote the emigration of Langerhans cells, which partly depends on the expression of mast-cell–derived IL-1β. We have previously shown that TLR7 ligation enhances transcutaneous immunization evoked by topical application of vaccine antigens to the skin, a procedure that directly targets skin-resident antigen-presenting cells. Consequently, we now demonstrate here that the capacity to mount a peptide-specific cytotoxic T-lymphocyte response following transcutaneous immunization using imiquimod as adjuvant is severely impaired in mast-cell–deficient mice. Thus, these findings demonstrate the potent versability of alternatively activated mast cells at the interface of innate and adaptive immunity.

The phenotype of NFATc2−/− c3−/− (double knockout [DKO]) mice implies a disturbed regulation of T cell responses, evidenced by massive lymphadenopathy, splenomegaly, and autoaggressive phenomena. The population of CD4+ CD25+ T cells from DKO mice lacks regulatory capacity, except a small subpopulation that highly expresses glucocorticoid-induced tumor necrosis factor receptor family–related gene (GITR) and CD25. However, neither wild-type nor DKO CD4+ CD25+ regulatory T cells (T reg cells) are able to suppress proliferation of DKO CD4+ CD25− T helper cells. Therefore, combined NFATc2/c3 deficiency is compatible with the development of CD4+ CD25+ T reg cells but renders conventional CD4+ T cells unresponsive to suppression, underlining the importance of NFAT proteins for sustaining T cell homeostasis.

Hearing loss is the most frequent long‐term complication of pneumococcal meningitis, affecting up to 40% of survivors. Unfortunately, adjuvant therapy with dexamethasone has failed to satisfactorily reduce its incidence. Therefore, we evaluated the use of antioxidants for the adjunctive therapy of meningitis‐associated deafness. Eighteen hours after intracisternal injection of 7.5 × 105colony‐forming units of Streptococcus pneumoniae, rats were treated systemically either with ceftriaxone and the antioxidants and peroxynitrite scavengers Mn(III)tetrakis(4‐benzoic acid)‐porphyrin (MnTBAP) or N‐acetyl‐L‐cysteine (NAC) or placebo (1ml phosphate‐buffered saline) for 4 days. Hearing was assessed by auditory brainstem response audiometry. Adjunctive antioxidant therapy significantly reduced the long‐term hearing loss (14 days after infection) for square wave impulses (mean hearing loss ± SD: ceftriaxone and placebo, 45±26dB; ceftriaxone and MnTBAP, 9±23dB; ceftriaxone and NAC, 19±30dB) as well as 1kHz (ceftriaxone and placebo, 28±19dB; ceftriaxone and MnTBAP, 10±16dB; ceftriaxone and NAC, 10±17dB), and 10kHz tone bursts (ceftriaxone and placebo, 62±27dB; ceftriaxone and MnTBAP, 16±13dB; ceftriaxone and NAC, 25±26dB). Furthermore, both antioxidants attenuated the morphological correlates of meningogenic hearing loss, namely, long‐term blood‐labyrinth barrier disruption, spiral ganglion neuronal loss, and fibrous obliteration of the perilymphatic spaces. Adjuvant antioxidant therapy is highly otoprotective in meningitis and therefore is a promising future treatment option.

In patients with acute bacterial meningitis, hearing loss can be transient but is often permanent. The mechanisms underlying meningitis‐associated hearing loss are not fully understood. Therefore, we investigated the morphological correlates of hearing loss in a rat model of pneumococcal meningitis. Transcutaneous intracisternal injection of Streptococcus pneumoniaeresulted in a dose‐dependent hearing loss (determined by auditory brainstem response audiometry), which was partially reversible during the acute stage. Nevertheless, a severe permanent hearing loss persisted until 2 weeks after infection. Suppurative labyrinthitis was accompanied by blood‐labyrinth barrier disruption (determined by cochlear Evans blue extravasation), which correlated closely with hearing loss during the acute stage but not after recovery. Two weeks after infection, spiral ganglion neuronal density was markedly decreased and correlated with the severity of permanent hearing loss. Neuronal loss can be explained by the new finding of meningitis‐associated spiral ganglion neuronal necrosis rather than apop‐tosis (determined by morphology, TUNEL staining, and immunohistochemistry).

In this study, the authors investigated the superficial multifidus muscle in patients with idiopathic scoliosis. During spinal fusion, biopsies were taken bilaterally at the apex of the curve, and at the upper and lower end vertebrae.

Objective: Oncogene alterations are thought to be prognostic indices in patients with breast cancer. The present study was carried out to investigate the amplification of the HER-2/neu and INT-2 oncogenes in ovarian cancer. Methods: In a retrospective study of 196 patients with epithelial ovarian cancer, the amplification of the oncogenes HER-2/neu and INT-2 in the DNA of paraffin-embedded tumor cells was determined by quantitative PCR. The purpose of this study was to analyze whether the two oncogenes correlated with such predictive factors as FIGO stage, histological grade, ascites, postoperative residual tumor mass, hormone receptor content, and preoperative CA 125 serum levels. The effect of HER-2/neu and INT-2 amplification on patient survival was also studied. Results: The only correlation found in this study was between INT-2 and preoperative CA 125 levels (P= 0.03). No correlations were demonstrable between HER-2/neu (log-rank test;P= 0.67) and INT-2 (log-rank test;P= 0.75) amplifications and overall survival. Conclusion: Unlike the established prognostic factors, neither HER-2/neu nor INT-2 appears to be predictive for survival in patients with ovarian cancer.

The Escherichia coli pdxC(serC) gene codes for a transaminase (EC 2.6.1.52). The gene is involved in both pyridoxine (vitamin B6) and serine biosynthesis and was over‐expressed as a MalE/PdxC(SerC) fusion protein. The fusion protein was purified by affinity chromatography on an amylose resin and hydrolyzed in the presence of protease factor Xa. Both the fusion protein and the PdxC(SerC) protein were characterized (KMvalue, turnover number, optimum pH). Both enzymes used 4‐O‐phosphoryl‐l‐threonine rather than 4‐hydroxy‐l‐threonine as a substrate indicating that the phosphorylated rather than the non‐phosphorylated amino acid is involved in pyridoxine biosynthesis. Pyridoxal phosphate was shown to be the cofactor for both enzymes and therefore seems to be involved in its own biosynthesis.

Posttranslational modification with SUMO is known to regulate the activity of transcription factors, but how SUMOylation of individual proteins might influence immunity is largely unexplored. The NFAT transcription factors play an essential role in antigen receptor-mediated gene regulation. SUMOylation of NFATc1 represses IL-2 in vitro, but its role in T cell–mediated immune responses in vivo is unclear. To this end, we generated a novel transgenic mouse in which SUMO modification of NFATc1 is prevented. Avoidance of NFATc1 SUMOylation ameliorated experimental autoimmune encephalomyelitis as well as graft-versus-host disease. Elevated IL-2 production in T cells promoted T reg expansion and suppressed autoreactive or alloreactive immune responses. Mechanistically, increased IL-2 secretion counteracted IL-17 and IFN-γ expression through STAT5 and Blimp-1 induction. Then, Blimp-1 repressed IL-2 itself, as well as the induced, proliferation-associated survival factor Bcl2A1. Collectively, these data demonstrate that prevention of NFATc1 SUMOylation fine-tunes T cell responses toward lasting tolerance. Thus, targeting NFATc1 SUMOylation presents a novel and promising strategy to treat T cell–mediated inflammatory diseases.

Data from untargeted metabolomics studies employing nuclear magnetic resonance (NMR) spectroscopy oftentimes contain negative values. These negative values hamper data processing and analysis algorithms and prevent the use of such data in multiomics integration settings. New methods to deal with such negative values are thus an urgent need in the metabolomics community. This study presents affine transformation of negative values (ATNV), a novel algorithm for replacement of negative values in NMR data sets. ATNV was implemented in the R package mrbin, which features interactive menus for user-friendly application and is available for free for various operating systems within the free R statistical programming language. The novel algorithms were tested on a set of human urinary NMR spectra and were able to successfully identify relevant metabolites.

Bronchoalveolar lavage (BAL) culture is a standard, though time-consuming, approach for identifying microorganisms in patients with severe lower respiratory tract infections. The sensitivity of BAL culture is relatively low, and prior antimicrobial therapy decreases the sensitivity further, leading to overuse of empirical antibiotics.

Whole-genome sequencing (WGS) is now routinely performed in clinical microbiology laboratories to assess isolate relatedness. With appropriately developed analytics, the same data can be used for prediction of antimicrobial susceptibility. We assessed WGS data for identification using open-source tools and antibiotic susceptibility testing (AST) prediction using ARESdb compared to matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identification and broth microdilution phenotypic susceptibility testing on clinical isolates from a multicenter clinical trial of the FDA-cleared Unyvero lower respiratory tract infection (LRTI) application (Curetis).

Tumor-necrosis-factor-alpha-induced protein 3 (TNFAIP3), or A20, is a ubiquitin-modifying protein and negative regulator of canonical nuclear factor κB (NF-κB) signaling. Several single-nucleotide polymorphisms in TNFAIP3are associated with autoimmune diseases, suggesting a role in tissue inflammation. While the role of A20 in peripheral immune cells has been well investigated, less is known about its role in the central nervous system (CNS). Here, we show that microglial A20 is crucial for maintaining brain homeostasis. Without microglial A20, CD8+T cells spontaneously infiltrate the CNS and acquire a viral response signature. The combination of infiltrating CD8+T cells and activated A20-deficient microglia leads to an increase in VGLUT1+terminals and frequency of spontaneous excitatory currents. Ultimately, A20-deficient microglia upregulate genes associated with the antiviral response and neurodegenerative diseases. Together, our data suggest that microglial A20 acts as a sensor for viral infection and a master regulator of CNS homeostasis.