Hermann J. Schluesener

Gold nanoparticle-based biosensors

The unique properties of gold nanoparticles have stimulated the increasing interest in the application of GNPs in interfacing biological recognition events with signal transduction and in designing biosensing devices exhibiting novel functions. The optical properties of GNPs provide wide range opportunities for construction optical biosensors. The excellent biocompatibility, conductivity, catalytic properties and high surface-to-volume ratio and high density of GNPs facilitate extensive application of GNPs in construction of electrochemical and piezoelectric biosensors with enhanced analytical performance with respect to other biosensor designs. In this article, the recent advances in construction of GNP-based optical, electrochemical and piezoelectric biosensors are reviewed, and some illustrative examples given, with a focus on the roles GNPs play in the biosensing process and the mechanism of GNPs for improving the analytical performances. Finally, the review concludes with an outline of present and future research for the real-world applications.

Natural polyphenols against neurodegenerative disorders: potentials and pitfalls.

Within the last years, a rapidly growing number of polyphenolic compounds with neuroprotective effects have been described. Many efforts have been made to explore the mechanisms behind the neuroprotective action of polyphenols. However, many pathways and mechanisms considered for mediating these effects are rather general than specific. Moreover, despite the beneficial effects of polyphenols in experimental treatment of neurodegeneration, little has been achieved in bringing them into routine clinical applications. In this review, we have summarized the protective effects of polyphenols against neurodegeneration, and we have also discussed some of the barricades in translating these biochemical compounds, into relevant therapeutics for neurodegenerative diseases.

Bone Morphogenetic Proteins: Neurotrophic Roles for Midbrain Dopaminergic Neurons and Implications of Astroglial Cells

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor β (TGF‐β) superfamily that have been implicated in tissue growth and remodelling. Recent evidence suggests that several BMPs are expressed in the developing and adult brain. Specifically, we show that BMP 2 and BMP 6 are expressed in the developing midbrain floor of the rat. We studied potential neurotrophic effects of BMPs on the in vitro survival, transmitter uptake and protection against MPP+ toxicity of mesencephalic dopaminergic neurons cultured from the embryonic midbrain floor at embryonic day (E) 14. At 10 ng/ml and under serum‐free conditions, most BMPs promoted the survival of dopaminergic neurons visualized by tyrosine hydroxylase immunocytochemistry during an 8‐day culture period, but to varying extents (relative potencies: BMP 6 = 12 > 2, 4, 7). BMPs 6 and 12 were as effective as fibroblast growth factor‐2 (FGF‐2) and glial cell line‐derived neurotrophic factor, promoting survival 1.7‐fold compared with controls. BMPs 9 and 11 were not effective. Dose‐response curves revealed an EC50 for BMPs 2, 6 and 12 of 2 ng/ml. BMPs 2, 4, 6, 7, 9 and 12 also promoted DNA synthesis and astroglial cell differentiation, visualized by 5‐bromodeoxyuridine (BrdU) incorporation and glial fibrillary acidic protein (GFAP) immunocytochemistry respectively. Suppression of cell proliferation and subsequent maturation of GFAP‐positive cells by 5‐fluorodeoxyuridine or aminoadipic acid abolished the neuron survival‐promoting effect of BMP 2. This suggests that BMPs, like other non‐TGF‐β factors affecting dopaminergic neuron survival, act indirectly, probably by stimulating the synthesis and/or release of glial‐derived trophic factors. BMP 6 and BMP 7 also increased the uptake of [3H]dopamine without affecting the uptake of [3H]5‐hydroxytryptamine and [3H]GABA, underscoring the specificity of the trophic effect. We conclude that several BMPs share a neurotrophic capacity for dopaminergic midbrain neurons with other members of the TGF‐β superfamily, but act indirectly, possibly through glial cells.

Long-term expression of heme oxygenase-1 (HO-1, HSP-32) following focal cerebral infarctions and traumatic brain injury in humans.

Abstract Extracellular heme derived from hemoglobin following hemorrhage or released from dying cells induces the expression of heme oxygenase-1 (HO-1, HSP-32) which metabolizes heme to the gaseous mediator carbon monoxide (CO), iron (Fe) and biliverdin. Biliverdin and its product bilirubin are powerful antioxidants. Thus, expression of HO-1 is considered to be a protective mechanism against oxidative stress and has been described in microglia, astrocytes and neurons following distinct experimental models of pathological alterations to the brain such as subarachnoidal hemorrhage, ischemia and traumatic brain injury (TBI) and in human neurodegenerative diseases. We have now analyzed the expression of HO-1 in human brains following TBI (n = 28; survival times: few minutes up to 6 months) and focal cerebral infarctions (FCI; n = 17; survival time: < 1 day up to months) by ¶immunohistochemistry. Follwing TBI, accumulation of ¶HO-1+ microglia/macrophages at the hemorrhagic lesion was detected as early as 6 h post trauma and was still pronounced after 6 months. In contrast, after FCI HO-1+ microglia/macrophages accumulated within focal hemorrhages only and were absent in non-hemorrhagic regions. Further, HO-1 was weakly expressed in astrocytes in the perifocal penumbra. In contrast to experimental data derived from rat focal ischemia, these results indicate a prolonged HO-1 expression in humans after brain injury.

Compound A, a Plant Origin Ligand of Glucocorticoid Receptors, Increases Regulatory T Cells and M2 Macrophages to Attenuate Experimental Autoimmune Neuritis with Reduced Side Effects

Experimental autoimmune neuritis (EAN) is a helper T cell-mediated autoimmune demyelinating inflammatory disease of the peripheral nervous system and serves as the animal model for human inflammatory demyelinating polyneuropathies. Compound A, a plant-derived phenyl aziridine precursor, was reported to activate glucocorticoid receptors to exert transrepression but not transactivation properties. In this study, we investigated the effects of Compound A in EAN rats. Compound A greatly suppressed paraparesis in EAN, even when administrated after the appearance of the first neurological signs. Accumulation of macrophages and lymphocytes, demyelination, and mRNA levels of inflammatory molecules in sciatic nerves of EAN were greatly attenuated by Compound A. In addition, Compound A inhibited progression of neuropathic pain and repressed microglia but not astrocyte activation and IL-1β and TNF-α up-regulation in EAN spinal cords. In EAN sciatic nerves, Compound A treatment increased numbers of anti-inflammatory M2 macrophages. Furthermore, Compound A induced the switch of macrophages from inflammatory M1 type to anti-inflammatory M2 type in vitro. In lymph nodes of EAN rats, Compound A depressed Th1 and Th17 cytokines, but increased Th2 cytokine and Foxp3 expression. An increase of Foxp3+/CD4+ regulatory T cells was seen in peripheral blood of EAN rats following Compound A treatment. In addition, Compound A did not cause a hyperglycemia effect in EAN rats as compared with the immunosuppressive steroid prednisolone. Therefore, our data demonstrated that Compound A could effectively suppress EAN with reduced side effects by attenuating inflammation, suggesting that Compound A could be a potent candidate for treatment of autoimmune neuropathies.

The allograft inflammatory factor-1 family of proteins

The allograft inflammatory factor‐1 (AIF‐1) is a 17 kDa interferon‐γ‐inducible Ca2+‐binding EF‐hand protein that is encoded within the HLA class III genomic region. Three proteins are probably identical with AIF‐1 termed Iba1 (ionized Ca2+‐binding adapter), MRF‐1 (microglia response factor) and daintain. Considerable but not complete sequence identity with AIF‐1 has been described for IRT‐1 (interferon‐responsive transcript), BART‐1 (balloon angioplasty‐responsive transcript), and other, yet unassigned alternatively spliced variants. In this review, genomic and functional characteristics of AIF‐1‐related proteins are summarized and a common nomenclature is proposed.

Dynamics of microglial activation after human traumatic brain injury are revealed by delayed expression of macrophage-related proteins MRP8 and MRP14

Abstract Human traumatic brain injury (TBI) is ideally suited for investigation of the kinetics of human microglial cell activation as the onset of lesion formation is precisely defined. The present study provides evidence of a distinct delay in macrophage/microglia response following TBI. Eighteen brains of patients who had survived TBI for 1 h to 6 months were analysed by immunohistology. Samples of contusional and non-contusional areas were studied using antibodies directed against antigens of microglia/ macrophages [major histocompatibility complex class II, CD4, interleukin (IL)-16, macrophage-related protein (MRP) 8 and MRP14]. IL-16, a natural ligand to CD4, was expressed constitutively by numerous microglial cells in all cases throughout the brain. CD4 could be detected regularly on perivascular cells. MRP8 and MRP14, which are only expressed on activated macrophages and microglial cells, could be detected only within brains with a survival time of more than 72 h post TBI. In addition, proliferation of microglia detected by MIB-1 was not present until 72 h. This delayed expression of the activation markers MRP8 and MRP14 and the proliferation marker MIB-1 is comparable to experimental closed head injuries but strictly different from acute activation found in ischemic brains.

Role of exosomes in immune regulation

Exosomes are small vesicles originating from late endosomes, 30–100 nm in diameter with typical cup‐shape morphology. They are reported to bear high levels of a narrow spectrum of molecules involved in immune response and signal transduction. Apart from removing obsolete membrane proteins, some surprising biological functions of exosomes were unveiled recently and their applications in immunotherapy of tumors are currently intensively investigated. Dendritic cell‐ (DC) and tumor‐derived exosomes have considerable anti‐tumor effects in experimental studies and several clinical trials. Despite their potential applications in eliciting a “positive” immune response, exosomes might induce some “unwanted” immune responses, such as immune tolerance and immune evasion. Therefore further investigations about the physiological functions of exosomes and the optimal way of exosome application in tumor immunotherapy are necessary. This review presents recent developments in the field of exosome research and focuses on its applications to tumor immunotherapy.

Spinal cord injury-induced lesional expression of the repulsive guidance molecule (RGM)

The repulsive guidance molecule (RGM) is involved in the formation of the central nervous system (CNS) during development by modulating guidance of growing axons. However, a role of RGM in CNS injury remains to be established. We studied the expression of RGM in the spinal cord of rats with spinal cord injury (SCI). After SCI, RGM+ cells accumulated in lesions and peri‐lesional areas. During the first days after SCI, RGM expression was confined to neurons, ballooned neurite fibers/retraction bulbs, smooth muscle/endothelial cells, and to leucocytes infiltrating the lesion. Lesional RGM expression was frequently confined to hypertrophic β‐APP+ and RhoA+ neurites/retraction bulbs. With maturation of the lesion, we observed RGM expression by components of the developing scar tissue (cicatrix), such as fibroblastoid cells, reactive astrocytes and in addition a pronounced extracellular RGM deposition resembling neo‐laminae. Frequent RGM+, RhoA+ coexpression by lesional retraction bulbs represent first preliminary evidence of RGM to exert growth inhibitory effects by the second messenger system RhoA. To date, RGM is one of the most potent axonal growth inhibitors identified and present in axonal growth impediments (i) oligodendrocytes; (ii) the plexus choroideus and (iii) components of the developing scar.

Allograft‐inflammatory factor‐1 in rat experimental autoimmune encephalomyelitis, neuritis, and uveitis: Expression by activated macrophages and microglial cells

Allograft inflammatory factor‐1 (AIF‐1) is a Ca2+binding peptide expressed predominantly by activated monocytes. In order to investigate the role of AIF‐1 in autoimmune lesions of the rat nervous system, we have used a synthetic gene to express AIF‐1 in E.coli and have produced monoclonal antibodies against AIF‐1.