Histopathological Study of Healing After Allogenic Mesenchymal Stem Cell Delivery in Myocardial Infarction in Dogs

In this histological study, we assessed the role of mesenchymal stem cells (MSCs) in the healing process that takes place during the subacute phase of myocardial infarction in dogs. Seven days after occlusion of the left anterior descending coronary artery, adult mongrel dogs received 100 × 10⁶ 4′-6-diamidino-2-phenylindole (DAPI)-labeled allogenic bone marrow–derived MSCs by the transendocardial (TE, n=6) and intracoronary (IC, n=4) routes; control dogs (n=6) received no infusion. The dogs were euthanized at 21 days after occlusion. Hearts were excised and sliced from apex to base into four transverse sections, which were divided into nine segments. Paraffin sections from each segment were stained with hematoxylin and eosin, trichrome, picrosirius red, and antibodies against several extracellular matrix components. Frozen sections were immunostained for host cardiac phenotypical markers and analyzed by epifluorescence and deconvolution fluorescence microscopy (DFM). We found less unresolved necrotic myocardium and more extracellular matrix deposition in MSC-treated dogs than in controls 2 weeks after cell delivery. By DFM, no DAPI+ MSC nuclei were observed within native cardiac cells. MSCs delivered during the subacute phase of acute myocardial infarction positively affect healing, apparently by mechanisms other than differentiation into mature native cardiac cells. (J Histochem Cytochem 57:167–176, 2009)     

Truncating loss-of-function mutations of <Emphasis Type="Italic">DISP1</Emphasis> contribute to holoprosencephaly-like microform features in humans

Defective function of the Sonic Hedgehog (SHH) signaling pathway is the most frequent alteration underlying holoprosencephaly (HPE) or its various clinical microforms. We performed an extensive mutational analysis of the entire human DISP1 gene, required for secretion of all hedgehog ligand(s) and which maps to the HPE 10 locus of human chromosome 1q41, as a HPE candidate gene. Here, we describe two independent families with truncating mutations in human DISP1 that resemble the cardinal craniofacial and neuro-developmental features of a recently described microdeletion syndrome that includes this gene; therefore, we suggest that DISP1 function contributes substantially to both of these signs in humans. While these clinical features are consistent with common HPE microforms, especially those linked to defective signaling by Sonic Hedgehog, we have insufficient evidence so far that functionally abnormal DISP1 alleles will commonly contribute to the more severe features of typical HPE.     

LPS-induced NF-κB expression in THP-1Blue cells correlates with neopterin production and activity of indoleamine 2,3-dioxygenase

Neopterin production is induced in human monocyte-derived macrophages and dendritic cells upon stimulation with Th1-type cytokine interferon-γ (IFN-γ). In parallel, IFN-γ induces the tryptophan-(trp)-degrading enzyme indoleamine 2,3-dioxygenase (IDO) and triggers the formation of reactive oxygen species (ROS). Translocation of the signal transduction element nuclear factor-κB (NF-κB) is induced by ROS and accelerates the pro-inflammatory response by activation of other pro-inflammatory pathways. Therefore, a close relationship between NF-κB expression, the production of neopterin and the degradation of trp can be assumed, although this has not been demonstrated so far. In the present in vitro study we compared the influence of lipopolysaccharide (LPS) on NF-κB activation, neopterin formation and the degradation of trp in THP-1Blue cells, which represent the human myelomonocytic cell line THP-1 stably transfected with an NF-κB inducible reporter system.In cells stimulated with LPS, a significant induction of NF-κB was observed, and this was paralleled by an increase of kynureunine (kyn) and neopterin concentrations and a decline of trp. The increase of the kyn to trp quotient indicates accelerated IDO activity. Higher LPS concentrations and longer incubation of cells were associated with higher activities of all three biochemical pathways and significant correlations existed between NF-κB activation, neopterin release and trp degradation (all p < 0.001). We conclude that there is a parallel induction of NF-κB, neopterin formation and trp degradation in monocytic THP-1 cells, which is elicited by pro-inflammatory triggers like LPS during innate immune responses.     

Online coupling of enantioselective capillary gas chromatography with proton nuclear magnetic resonance spectroscopy

The hyphenation of enantioselective capillary gas chromatography and mass spectrometry is not always sufficient to distinguish between structural isomers, thus requiring peak identification by NMR spectroscopy. Here the first online coupling of enantioselective capillary gas chromatography with proton nuclear resonance spectroscopy is described for the unfunctionalized chiral alkane 2,4‐dimethylhexane resolved on octakis(6‐O‐methyl‐2,3‐di‐O‐pentyl)‐γ‐cyclodextrin at 60°C. NMR allows constitutional and configurational isomers (diastereomers and enantiomers) to be distinguished. Enantiomers display identical spectra at different retention times, which enable an indirect identification of these unfunctionalized alkanes. The presented method is still at an early development stage, and will require instrumental optimization in the future. Chirality 2010. © 2010 Wiley‐Liss, Inc.     

Toward a better understanding of ADHD: LPHN3 gene variants and the susceptibility to develop ADHD

During the past 15 years, an impressive amount of genetic information has become available in the research field of psychiatry, particularly as it relates to attention-deficit/hyperactivity disorder (ADHD). However, the classical clinical approach to ADHD has minimally affected and not significantly been improved by this genetic revolution. It is difficult to predict how long it will take for genetic findings to alter the way clinicians treat patients with ADHD. New medications or treatment protocols may take years to become routine clinical practice. However, when taken together, recent successes in genomics, pharmacogenomics, and genetic epidemiology have the potential (1) to prevent comorbid consequences of ADHD, (2) to individualize therapies for patients with ADHD, and (3) to define new epidemiological policies to aid with the impact of ADHD on society. Here, we present an overview of how genetic research may affect and improve the quality of life of patients with ADHD: as an example, we use the discovery of LPHN3, a new gene in which variants have recently been shown to be associated with ADHD.     

Die Lipide von Endomycopsis vernalis bei verschiedener Stickstoff-Ernährung

1. Endomycopsis vernalis was cultivated on media with different N supply: series A 1%, series B 0,125% asparagine. Sonified cells were extracted and yielded 14.3% (A) and 65.3 (B) total lipids/non lipid dry matter respectively.
2. Neutral and complex lipids were separated by rubber membrane dialysis. There is no difference in the percentage of complex lipids of both series. The increase of lipids in cells grown on low N level is due to a higher content of neutral lipids.
3. Components of the neutral lipids, analysed by DC, were diglycerides, triglycerides, free and esterified ergosterol. Their percentage is influenced by the nutritional conditions. There is a significant increase of triglycerides and of sterol esters in the high lipid cells of series B.
4. Methyl esters of component fatty acids of glycerides and sterol esters were analyzed by GLC. Saturated acids C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, monoenic acids C₁₆ and C₁₈, linoleic and linolenic acids were found to be present. Major acids were in all cases 18:1 (17–57%), 18:2 (18–50%) and 16:0 (10–18%). Linolenic acid is higher in di-and triglycerides of low lipid cells of series A than in high lipid cells of series B. Both qualitative and quantitative differences of fatty acids were found in sterol esters of series A and B respectively.
5. The major components of complex lipids, identified by DC and isolated by CC, in both series, were phosphatidyl choline (A:36.5, B:41.0%) and phosphatidyl ethanolamine (A:24.9, B:20.5%) in addition to small amounts of lysophosphatidyl choline, lysophosphatidyl ethanolamine, phosphatidyl serine, monophosphoinositide, diphosphatidyl glycerol and, possibly cerebroside like substances.
6. Methyl esters of the fatty acids of phosphatidyl choline and ethanolamine from both series were determined by GLC. In all samples 16:0, 18:0, 18:1, 18:2 and 18:3 acids were present besides of traces of 16:1 and 17:0. In contrast to neutral lipids the major acid of phospholipids is linoleic (53–58%), followed by oleic (8–24%) and linolenic acid (1–18%). The percentages of palmitic (4–8%) and stearic acids (tr.-1%) are small. Low lipid cells of series A differ from high lipid cells of series B by an increase of linolenic, and a decrease of linoleic acids, both in phosphatidyl choline and phosphatidyl ethanolamine.