Signs of immaturity of splenic dendritic cells from the autoimmune prone biobreeding rat: consequences for the in vitro expansion of regulator and effector T cells

From the biobreeding-diabetic prone (BB-DP) rat, an animal model for endocrine autoimmunity, phenotype and function of splenic dendritic cells (DC) were studied. Furthermore, the suppressive effect of peritoneal macrophages (pMphi) from the BB-DP rat in the MLR was investigated. Lower numbers of splenic DC were isolated from BB-DP rats than from control Wistar rats. In the preautoimmune phase, DC of the BB-DP rat had a lower surface MHC class II expression (and in preliminary data, a lower CD80 expression), ingested more bacteria, and had a lower stimulatory potency in the syngeneic (syn)MLR as compared with control DC. During disease development, the MHC class II expression further decreased, and a low stimulatory activity became evident in the allogeneic (allo)MLR. With regard to the expansion of suppressor/regulatory T cells, a lower percentage of RT6+ T cells but higher percentages of CD45RClow T cells were induced by BB-DP DC in synMLR, but not in alloMLR. An increase in the CD4/CD8 T cell ratio was observed in both the syn- and alloMLR due to a relative weak expansion of CD8+ T cells with DC of the BB-DP rat. Resident pMphi isolated from BB-DP or Wistar rats were equally effective in suppressing the DC-driven synMLR. In conclusion, splenic DC from the BB-DP rat have a lower accessory cell function already at young age, before the development of disease, and expanded different subsets of effector/suppressor T cells in vitro as compared with those from Wistar rats. The dysfunction of DC from BB-DP rats is likely to be caused by their relative immaturity as indicated by their low class II and costimulatory molecule expression and relatively high phagocytic activity

Multisite monitoring of choline using biosensor microprobe arrays in combination with CMOS circuitry

A miniature device enabling parallel in vivo detection of the neurotransmitter choline in multiple brain regions of freely behaving rodents is presented. This is achieved by combining a biosensor microprobe array with a custom-developed CMOS chip. Each silicon microprobe comprises multiple platinum electrodes that are coated with an enzymatic membrane and a permselective layer for selective detection of choline. The biosensors, based on the principle of amperometric detection, exhibit a sensitivity of 157±35 µA mM-1 cm-2, a limit of detection of below 1 µM, and a response time in the range of 1 s. With on-chip digitalization and multiplexing, parallel recordings can be performed at a high signal-to-noise ratio with minimal space requirements and with substantial reduction of external signal interference. The layout of the integrated circuitry allows for versatile configuration of the current range and can, therefore, also be used for functionalization of the electrodes before use. The result is a compact, highly integrated system, very convenient for on-site measurement

Differences in renal hemodynamics and renin secretion between patients with unifocal and multifocal fibromuscular dysplasia

Objective: Fibromuscular dysplasia (FMD) can be classified in a multifocal and a unifocal subtype. As unifocal FMD generally leads to more severe hypertension at younger age, we hypothesized that renal hemodynamics are more disturbed in unifocal renal artery FMD as compared with multifocal FMD, leading to increased renin secretion. Methods: We measured renal blood flow ((133)Xenon washout method), renin secretion, and glomerular filtration rate per kidney in 101 patients with FMD (26 unifocal and 75 multifocal), all off medication and prior to balloon angioplasty. Results: We found that renal blood flow and glomerular filtration were substantially lower in kidneys with unifocal FMD as compared with multifocal FMD. In the affected kidney from patients with unilateral FMD for example, mean renal blood flow was 173 +/- 77 in unifocal vs. 244 +/- 79 ml/100 g kidney/min in multifocal FMD (P=0.013). Moreover, lateralization in renin secretion was only observed in a subset of patients with unifocal FMD, but not in any of the patients with multifocal FMD. Conclusion: These findings suggest that the impact of unifocal FMD lesions on the kidney is more severe, resulting in a classical pattern of renovascular hypertension. In multifocal FMD, however, renal blood flow is more preserved, local renin secretion is not increased, and the association between renin levels and blood pressure is inverse. These differences may explain the often more severe clinical presentation and higher success rate of revascularization in unifocal FMD, but also suggest that the pathophysiological mechanisms leading to hypertension may differ between these two disease entities

Mutation and drug-specific intracellular accumulation of EGFR predict clinical responses to tyrosine kinase inhibitors

Background: Clinical responses to EGFR tyrosine kinase inhibitors (TKIs) are restricted to tumors harboring specific activating mutations and even then, not all tyrosine kinase inhibitors provide clinical benefit. All TKIs however, effectively inhibit EGFR phosphorylation regardless of the mutation present. Methods: High-throughput, high-content imaging analysis, western blot, Reversed phase protein arrays, mass spectrometry and RT-qPCR. Findings: We show that the addition of TKIs results in a strong and rapid intracellular accumulation of EGFR. This accumulation mimicked clinical efficacy as it was observed only in the context of the combination of a TKI-sensitive mutation with a clinically effective (type I) TKI. Intracellular accumulation of EGFR was able to predict response to gefitinib in a panel of cell-lines with different EGFR mutations. Our assay also predicted clinical benefit to EGFR TKIs on a cohort of pulmonary adenocarcinoma patients (hazard ratio 0.21, P=0.0004 [Cox proportional hazard model]) and could predict the clinical response in patients harboring rare mutations with unknown TKI-sensitivity. All investigated TKIs, regardless of clinical efficacy, inhibited EGFR phosphorylation and downstream pathway activation, irrespective of the mutation present. Intracellular accumulation of EGFR depended on a continued presence of TKI indicating (type I) TKIs remain associated with the protein even after its dephosphorylation. Accumulation therefore is likely caused by two consecutive conformational changes, induced by both activating mutation and TKI, that combined block EGFR-membrane recycling. Interpretation: We report on an assay that mimics the discrepancy between molecular and clinical activity of EGFR-TKIs, which may allow response prediction in vitro and helps understand the mechanism of effective inhibitors

Magnetic nanodoping: Atomic control of spin states in cobalt doped silver clusters

The interaction of magnetic dopants with delocalized electron states can result in interesting many-body physics. Here, the magnetic properties of neutral and charged finite silver metal host clusters with a magnetic cobalt atom impurity were investigated experimentally by exploiting the complementary methods of Stern- Gerlach molecular beam deflection and x-ray magnetic circular dichroism spectroscopy and are accompanied by density functional theory calculations and charge transfer multiplet simulations. The influence of the number of valence electrons and the consequences of impurity encapsulation were addressed in free size-selected, singly cobalt-doped silver clusters CoAg0,n+ (n = 2–15). Encapsulation of the dopant facilitates the formation of delocalized electronic shells with complete hybridization of the impurity 3d- and the host 5s-derived orbitals, which results in impurity valence electron delocalization, effective spin relaxation, and a low-spin ground state. In the exohedral size regime, spin pairing in the free electron gas formed by the silver 5s electrons is the dominating driving force determining the local 3d occupation of the impurity and therefore, adjusting the spin magnetic moment accordingly

Multisite monitoring of choline using biosensor microprobe arrays in combination with CMOS circuitry

A miniature device enabling parallel in vivo detection of the neurotransmitter choline in multiple brain regions of freely behaving rodents is presented. This is achieved by combining a biosensor microprobe array with a custom-developed CMOS chip. Each silicon microprobe comprises multiple platinum electrodes that are coated with an enzymatic membrane and a permselective layer for selective detection of choline. The biosensors, based on the principle of amperometric detection, exhibit a sensitivity of 157 +/- 35 mu A mM(-1) cm(-2), a limit of detection of below 1 mu M, and a response time in the range of 1 s. With on-chip digitalization and multiplexing, parallel recordings can be performed at a high signal-to-noise ratio with minimal space requirements and with substantial reduction of external signal interference. The layout of the integrated circuitry allows for versatile configuration of the current range and can, therefore, also be used for functionalization of the electrodes before use. The result is a compact, highly integrated system, very convenient for on-site measurements

Greener pulmonary care is possible

Bij het gebruik van dosisaerosolinhalatoren door COPD- en astmapatiënten komen drijf­gassen vrij die bijdragen aan de opwarming van de aarde. Wanneer we dit type inhalatoren bij 70% van de 1,4 miljoen gebruikers veilig vervangen door klimaatvriendelijkere poeder­inhalatoren, kan de uitstoot van broeikasgassen met ongeveer 63 miljoen kg CO2-eq per jaar afnemen. Een behoorlijke milieuwinst. De goedkoopste vervangers leveren per jaar bovendien een flinke kostenbesparing op. Wij denken dat de milieu-impact in de longzorg aanzienlijk naar beneden kan

Greener pulmonary care is possible

Bij het gebruik van dosisaerosolinhalatoren door COPD- en astmapatiënten komen drijf­gassen vrij die bijdragen aan de opwarming van de aarde. Wanneer we dit type inhalatoren bij 70% van de 1,4 miljoen gebruikers veilig vervangen door klimaatvriendelijkere poeder­inhalatoren, kan de uitstoot van broeikasgassen met ongeveer 63 miljoen kg CO2-eq per jaar afnemen. Een behoorlijke milieuwinst. De goedkoopste vervangers leveren per jaar bovendien een flinke kostenbesparing op. Wij denken dat de milieu-impact in de longzorg aanzienlijk naar beneden kan.Bij het gebruik van dosisaerosolinhalatoren door COPD- en astmapatiënten komen drijf­gassen vrij die bijdragen aan de opwarming van de aarde. Wanneer we dit type inhalatoren bij 70% van de 1,4 miljoen gebruikers veilig vervangen door klimaatvriendelijkere poeder­inhalatoren, kan de uitstoot van broeikasgassen met ongeveer 63 miljoen kg CO2-eq per jaar afnemen. Een behoorlijke milieuwinst. De goedkoopste vervangers leveren per jaar bovendien een flinke kostenbesparing op. Wij denken dat de milieu-impact in de longzorg aanzienlijk naar beneden kan