an-vitro-hemodynamic-loop-model-to-investigate-hemocytocompatibility

An In Vitro Hemodynamic Loop Model to Investigate the Hemocytocompatibility and Host Cell Activation of Vascular Medical Devices

In this study, the hemocompatibility of tubes with an inner diameter of 5 mm made of polyvinyl chloride (PVC) and coated with different bioactive ...

To compare markers of alcohol consumption.

Determination of glucuronidated 5-hydroxytryptophol (GTOL), a marker of recent alcohol intake, by ELISA technique.

Endothelial function as determined by flow-mediated dilation (FMD) deteriorates during postprandial lipemia. The impairment can be neutralized by adding 50 g casein to the fatty meal. The objective of this study was to see which of the casein constituents is responsible for this neutralization effect.

Addition of 2.5 g L-arginine in a fatty meal prevents the lipemia-induced endothelial dysfunction in healthy volunteers.

Polymorphonuclear neutrophils (PMN) mediate early immunity to infection but can also cause host damage if their effector functions are not controlled. Their lack or dysfunction is associated with severe health problems and thus the analysis of PMN physiology is a central issue. One prerequisite for PMN analysis is the availability of purified cells from primary organs. While human PMN are easily isolated from peripheral blood, this approach is less suitable for mice due to limited availability of blood. Instead, bone marrow (BM) is an easily available reservoir of murine PMN, but methods to obtain pure cells from BM are limited. We have developed a novel protocol allowing the isolation of highly pure untouched PMN from murine BM by negative immunomagnetic isolation using a complex antibody cocktail. The protocol is simple and fast (~1 h), has a high yield (5-10*10â¶ PMN per animal) and provides a purity of cells equivalent to positive selection (>80%). Most importantly, cells obtained by this method are non-activated and remain fully functional in vitro or after adoptive transfer into recipient animals. This method should thus greatly facilitate the study of primary murine PMN in vitro and in vivo.

Rapid immunomagnetic negative enrichment of neutrophil granulocytes from murine bone marrow for functional studies in vitro and in vivo.

Neutrophilen Neutrophilen (PMN) fr&uuml;h gegen die Infektion zu vermitteln, sondern k&ouml;nnen auch Host-Sch&auml;den verursachen, wenn ihre Effektor-Funktionen nicht gesteuert werden. Ihre Mangel oder Funktionsst&ouml;rung ist verkn&uuml;pft mit schweren gesundheitlichen Problemen und so die Analyse von PMN Physiologie ist ein zentrales Thema. Eine Voraussetzung f&uuml;r die Analyse von PMN ist die Verf&uuml;gbarkeit von gereinigter Zellen von prim&auml;ren Organe. W&auml;hrend menschliche PMN sind leicht isoliert aus peripherem Blut, ist dieser Ansatz weniger geeignet f&uuml;r M&auml;use aufgrund der begrenzten Verf&uuml;gbarkeit von Blut. Stattdessen Knochenmark (BM) ist eine leicht zug&auml;ngliche Reservoir von murinen PMN, aber reine Zellen von BM erhalten sind beschr&auml;nkt. Wir haben ein neuartiges Protokoll &uuml;ber die Isolierung von hochreinen unber&uuml;hrte PMN von murinen BM von negativen Immunomagnetic-Isolation, die mit einen komplexen Antik&ouml;rper-Cocktail entwickelt. Das Protokoll ist einfach und schnell (~ 1 h), hat eine hohe Rendite (5-10 * 10&#x2076; PMN pro Tier) und bietet eine Reinheit der Zellen entspricht positive Auswahl (&gt; 80 %). Vor allem Zellen, die von dieser Methode sind, nicht aktivierte und bleiben voll funktionsf&auml;hig in Vitro oder nach adoptiven Transfer in Empf&auml;nger Tiere. Diese Methode sollte das Studium der prim&auml;ren murinen PMN in vitro und in vivo somit erheblich erleichtern.

Schnelle Immunomagnetic negative Bereicherung der neutrophile Granulozyten aus murinen Knochenmark f&uuml;r Funktionsstudien in vitro und in vivo.

DCs contribute to immune homeostasis under physiological conditions and regulate the immune activation during infection. The deubiquitinase A20 inhibits the activation of NF-κB-dependent immune reactions, and prevents the hyperactivation of DCs under steady-state conditions. However, the role of DC-specific A20 under pathological conditions is unknown. Here, we demonstrate that upon injection of low-dose LPS, mice with DC-specific A20 deletion (CD11c-Cre A20(fl/fl) ) died within 6 h, whereas A20(fl/fl) controls survived. LPS-induced mortality in CD11c-Cre A20(fl/fl) mice was characterized by increased serum levels of IL-2, IL-10, IL-12, IFN-γ, and TNF. Upon LPS stimulation, the activation of NF-κB and ERK-NFATc3 pathways were enhanced in A20-deficient DCs, resulting in an increased production of IL-2, IL-12, and TNF both in vitro and in vivo. Targeted inhibition of ERK in A20-deficient DCs abolished the increased production of IL-2. A20-deficient DCs failed to induce LPS tolerance, which was independent of T cells and the intestinal flora, since T-cell depletion and decolonization of CD11c-Cre A20(fl/fl) mice could not prevent death of LPS-challenged CD11c-Cre A20(fl/fl) mice. In conclusion, these findings show that DC-specific A20 preserves immune homeostasis in steady-state conditions and is also required for LPS tolerance.

A20 expression in dendritic cells protects mice from LPS-induced mortality.

Anemia is a common presentation in children but the differential diagnosis of iron deficiency and β-thalassemia remains a diagnostic challenge. Red blood cell indices have been shown to perform weakly in such scenarios. One potential cause is breastfeeding, but the evidence for unusually prolonged exclusive breastfeeding as a cause of iron deficiency anemia in older (>2 years) toddlers is sparse and the association of breastfeeding with iron deficiency in this age group of older toddlers is not unequivocally established. In this case we describe an unusual cause of nutritional iron deficiency anemia in the age group of 2-3 years.

A two-and-a-half-year-old breastfed toddler presenting with anemia: a case report.

Cholesterol is reduced in depressed patients, however, these patients have a higher risk for cardiovascular diseases. Electroconvulsive therapy (ECT) is a highly effective treatment option for specific forms of depression. Like for other non-pharmacological therapies targeting depression such as psychotherapy or sleep deprivation, there is a lack of evidence about the effects on peripheral lipid parameters. Our objective was to study the impact of ECT as a non-pharmacological treatment on the peripheral lipid pattern in depressive patients.

Serum lipid profile changes after successful treatment with electroconvulsive therapy in major depression: A prospective pilot trial.

Bile acids (BAs) are increasingly recognised as metabolic regulators, potentially improving insulin sensitivity following bariatric surgery. However, physiological relevance of such observations remains unknown. Hence, we analysed serum BA composition and associated gut-derived hormone levels following lifestyle-induced weight loss in individuals with metabolic syndrome (MetS). 74 non-smoking men (45-55 yr) with MetS were randomised to a lifestyle-induced weight loss program (supervision via telemonitoring) or to a control arm. Before and after a 6 months intervention period clinical and laboratory parameters, body composition, serum BA profile, FGF-19, and GLP-1 concentrations were determined in fasting blood samples. 30 participants in the control and 33 participants in the treatment arm completed the study and were included in the data analysis. In participants of the treatment arm lifestyle-induced weight loss resulted in markedly improved insulin sensitivity. Serum levels of BA species and total GLP-1 decreased, while FGF-19 remained stable. Serum BA composition changed towards an increased 12α-hydroxylated/non-12α-hydroxylated ratio. None of these parameters changed in participants of the control arm. Our results demonstrate that improved metabolic control by lifestyle modifications lowers serum levels of BAs and GLP-1 and changes serum BA composition towards an increased 12α/non-12α ratio (ICTRP Trial Number: U1111-1158-3672).

Serum bile acids and GLP-1 decrease following telemetric induced weight loss: results of a randomized controlled trial.

Autosomal-dominant familial hypercholesterolemia (FH) is characterized by elevated plasma levels of low-density lipoprotein cholesterol (LDL-C) and a dramatically increased risk to develop cardiovascular disease (CVD). Mutations in three major genes have been associated with FH: the LDL receptor gene (LDLR), the apolipoprotein B gene (APOB), and the proprotein convertase subtilisin/kexin 9 gene (PCSK9). Here we investigated the frequency and the spectrum of FH causing mutations in Germany.

Clinical characterization and mutation spectrum of German patients with familial hypercholesterolemia.

Circulating proteins are widely used as biomarkers in clinical applications for the diagnosis, prediction, and treatment of numerous diseases. Immunoassays are the most common technologies for quantification of protein biomarkers and exist in various formats. Traditional immunoassays offer sensitive and fast analyses but cannot differentiate between proteoforms. Protein microheterogeneity, mainly due to post-translational modification, has been recognized as a fingerprint for different pathologies, and knowledge about proteoforms is an important step toward personalized medicine. Mass spectrometry (MS) has emerged to be a powerful technique for the characterization and quantification of proteoforms. We have established a novel four-plex immunoassay based on Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) MS for the targeted quantification of the inflammatory markers C-reactive protein (CRP), serum amyloid A (SAA), and calprotectin (S100A8/9) and the kidney function marker cystatin C (CysC). Antibodies were covalently bound to superparamagnetic beads, which delivered robust and fast sample processing. Polyhistidine-tagged recombinant target proteins were used as internal standards for quantification. Our method identified a number of proteoforms for SAA ( n = 11), S100A8/9 ( n = 4) and CysC ( n = 4). The assay was characterized by low limits of detection (0.01-0.06 μg/mL) and low coefficients of variation (3.8-9.4%). Method validation demonstrated good between-assay agreement with immuno-turbidimetry ( R = 0.963 for CRP), ELISA ( R = 0.958 for SAA; R = 0.913 for S100A8/9), and nephelometry ( R = 0.963 for CysC). The low sample consumption of 20 μL and the high sample throughput of 384 samples per day make this targeted immuno-MALDI approach suited for assessment of inflammatory and renal status in large cohort studies based on precious biobanks samples.

Multiplex Immuno-MALDI-TOF MS for Targeted Quantification of Protein Biomarkers and Their Proteoforms Related to Inflammation and Renal Dysfunction.

Innate immunity has been linked to initiation of Alzheimer's disease and multiple sclerosis. Moreover, risk of first-episode psychosis (FEP) and schizophrenia (Sz) is increased after various infections in predisposed individuals. Thus, we hypothesized an analogous role of innate immunity with increased C-reactive protein (CRP) in non-affective psychosis. Differential blood count, CRP, neutrophil and monocyte-macrophage activation markers, cortisol and psychotic symptoms (Positive and Negative Syndrome Scale [PANSS]) were assessed in controls (n = 294) and acutely ill unmedicated FEP (n = 129) and Sz (n = 124) patients at baseline and after 6 weeks treatment. Neutrophils, monocytes, and CRP were increased in patients vs controls at baseline (P < .001), and neutrophil and monocyte counts correlated positively with activation markers. Eosinophils were lower at baseline in FEP (P < .001) and Sz (P = .021) vs controls. Differences in neutrophils (P = .023), eosinophils (P < .001), and CRP (P < .001) were also present when controlling for smoking and cortisol, and partially remitted after antipsychotic treatment. FEP patients with high neutrophils (P = .048) or monocytes (P = .021) had higher PANSS-P scores at baseline but similar disease course. CRP correlated with PANSS-P at baseline (ρ = 0.204, P = .012). Improvement of positive symptoms after treatment correlated with declining neutrophils (ρ = 0.186, P = .015) or CRP (ρ = 0.237, P = .002) and rising eosinophils (ρ = -0.161, P = .036). In FEP, normalization of neutrophils (ρ = -0.231, P = .029) and eosinophils (ρ = 0.209, P = .048) correlated with drug dosage. In conclusion, innate immune system activation correlated with PANSS-P, supporting the immune hypothesis of psychosis. Neutrophil and monocyte counts and CRP levels may be useful markers of disease acuity, severity, and treatment response.

Innate Immune Cells and C-Reactive Protein in Acute First-Episode Psychosis and Schizophrenia: Relationship to Psychopathology and Treatment.

Disturbed iron (Fe) ion homeostasis and mitochondrial dysfunction have been implicated in neurodegeneration. Both processes are related, because central Fe ion consuming biogenetic pathways take place in mitochondria and affect their oxidative energy metabolism. Iron is imported into mitochondria by the two homologous Fe ion importers mitoferrin-1 and mitoferrin-2. To elucidate more specifically the role of mitochondrial Fe ions for brain energy metabolism and for proper neuronal function, we generated mice with a neuron-specific knockout of mitoferrin-1 (Slc25a37 or mfrn-1) and compared them with corresponding control littermates (mfrn-1). Mice lacking neuronal mfrn-1 exhibited no obvious anatomical or behavioral abnormalities as neonates, young or adult animals. However, they exhibited a moderate decrease in brain mitochondrial O-consumption with complex-I substrates of the electron transport chain (p < 0.05), indicating a moderate suppression of electron transport. While these mice did not exhibit altered basal fear levels, inquisitiveness or motor skills in specific neurobiological test batteries, they clearly exhibited decreased spatial learning skills and missing establishment of stable spatial memory in Morris water maze, as compared to floxed controls (p < 0.05). We thus conclude that mitochondrial Fe ion supply is an important player in neuronal energy metabolism and proper brain function and that the carrier mitoferrin-1 cannot be completely replaced by mitoferrin-2 or other as yet unknown Fe ion carriers.

Article	Year
Determination of glucuronidated 5-hydroxytryptophol (GTOL), a marker of recent alcohol intake, by ELISA technique. Clinical biochemistry\| PubMed ID: 17112495	2007
Addition of 2.5 g L-arginine in a fatty meal prevents the lipemia-induced endothelial dysfunction in healthy volunteers. Atherosclerosis\| PubMed ID: 19081092	2009
Rapid immunomagnetic negative enrichment of neutrophil granulocytes from murine bone marrow for functional studies in vitro and in vivo. PloS one\| PubMed ID: 21383835	2011
A20 expression in dendritic cells protects mice from LPS-induced mortality. European journal of immunology\| PubMed ID: 25472594	2015
A two-and-a-half-year-old breastfed toddler presenting with anemia: a case report. BMC research notes\| PubMed ID: 25514884	2014
Serum lipid profile changes after successful treatment with electroconvulsive therapy in major depression: A prospective pilot trial. Journal of affective disorders\| PubMed ID: 26426831	2016
Serum bile acids and GLP-1 decrease following telemetric induced weight loss: results of a randomized controlled trial. Scientific reports\| PubMed ID: 27452603	2016
Clinical characterization and mutation spectrum of German patients with familial hypercholesterolemia. Atherosclerosis\| PubMed ID: 27596133	2016
Multiplex Immuno-MALDI-TOF MS for Targeted Quantification of Protein Biomarkers and Their Proteoforms Related to Inflammation and Renal Dysfunction. Analytical chemistry\| PubMed ID: 29420882	2018
Innate Immune Cells and C-Reactive Protein in Acute First-Episode Psychosis and Schizophrenia: Relationship to Psychopathology and Treatment. Schizophrenia bulletin\| PubMed ID: 31504969	2020
Mitoferrin-1 is required for brain energy metabolism and hippocampus-dependent memory. Neuroscience letters\| PubMed ID: 31563673	2019

Katrin Borucki

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