Exploring quantitative cellular bioimaging and assessment of CdSe/ZnS quantum dots cellular uptake in single cells, using ns-LA-ICP-SFMS.

Quantitative bioimaging of Quantum Dots (QDs) uptake in single cells by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is a challenging task due to the high sensitivity and high spatial resolution required, and to the lack of matrix-matched reference materials. In this work, high spatially resolved quantitative bioimaging of CdSe/ZnS QDs uptake in single HT22 mouse hippocampal neuronal cells and in single HeLa human cervical carcinoma cells is novelty investigated combining: (a) the use of a ns-LA-ICP-Sector Field (SF)MS unit with mono-elemental fast and sensitive single pulse response for 114Cd+; and (b) the spatially resolved analysis of dried pL-droplets from a solution with a known concentration of these QDs to obtain a response factor that allows quantification of elemental bioimages. Single cells and dried pL-droplets are morphologically characterized by Atomic Force Microscopy (AFM) to determine their volume and thickness distribution. Moreover, operating conditions (e.g. spot size, energy per laser pulse, etc.) are optimized to completely ablate the cells and pL droplets at high spatial resolution. Constant operating conditions for the analysis of the single cells and calibrating samples is employed to reduce potential fractionation effects related to mass load effects in the ICP. A number concentration of CdSe/ZnS QDs between 3.5 104 and 48 104 is estimated to be uptaken by several selected single HT22 and HeLa cells, after being incubated in the presence of a QDs suspension added to a standard cell culture medium. Mono-elemental bioimaging at subcellular resolution seems to show a higher number concentration of the CdSe/ZnS QDs in the cytosol around the cell nucleus.

Properties of in situ generated gold nanoparticles in the cellular context.

Gold nanostructures that serve as probes for nanospectroscopic analysis of eukaryotic cell cultures can be obtained by the in situ reduction of tetrachloroauric acid (HAuCl4). To understand the formation process of such intracellularly grown particles depending on the incubation medium, the reaction was carried out with 3T3 fibroblast cells in three different incubation media, phosphate buffer, Dulbecco's Modified Eagle Medium (DMEM), and standard cell culture medium (DMEM with fetal calf serum). The size, the optical properties, the biomolecular corona, and the localization of the gold nanoparticles formed in situ vary for the different conditions. The combination of surface-enhanced Raman scattering (SERS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) microscopic mapping and transmission electron microscopy (TEM) provides complementary perspectives on plasmonic nanoparticles and non-plasmonic gold compounds inside the cells. While for the incubation with HAuCl4 in PBS, gold particles provide optical signals from the nucleus, the incubation in standard cell culture medium leads to scavenging of the toxic molecules and the formation of spots of high gold concentration in the cytoplasm without formation of SERS-active particles inside the cells. The biomolecular corona of nanoparticles formed in situ after incubation in buffer and DMEM differs, suggesting that different intracellular molecular species serve for reduction and stabilization. Comparison with data obtained from ready-made gold nanoparticles suggests complementary application of in situ and ex situ generated nanostructures for optical probing.

[Forensic psychiatry in Germany. Comparison of different trends of expansion in the different states of Germany]. / Massregelvollzug nach para. 63 StGB. Vergleich der Entwicklungen in den Bundesländern.

BACKGROUND: German forensic psychiatric hospitals are confronted with a continuously increasing number of mentally disordered offenders. It has not yet been studied whether and to what extent this increase varies between individual states of the German republic. Variability would indicate possibilities of regulation. METHODS: We analyzed annual data from the Federal German Office of Statistics on hospital treatment orders (Paragraph 63 German penal code; incidence) and capacity/occupancy of forensic hospitals (prevalence). The observation period included the period from 1993 to 2003. RESULTS: There were noticeable differences in all cross-sectional and longitudinal measures (rates of sentencing/incidence and occupancy/prevalence, slope of incidence growth and prevalence, rates of balancing/discharging). Differences between the federal states increased over the period under observation. CONCLUSIONS: Divergent trends of the German states despite common statutory foundations are indicative of options to control the increase in mentally disordered offenders. Possible causes of the divergent trends are discussed.

Living donor kidney transplantation from the elderly donor.

PURPOSE: The organ shortage has led to increasing acceptance of living donation in all transplant centers. Although the risk of impaired long-term outcome seems to be greater using elderly donors, these organs are not generally refused for transplantation. We report our experience with 25 living donor kidney transplantations from donors older than 60 years. METHODS: Between 1995 and 2004, 124 living donor procedures were performed in our center from 83 related and 41 unrelated donors. Twenty-five donors (19 female, 6 male) were 60 years or older (mean, 65.3 +/- 3.9 years). The recipient included (10 females and 15 males) showed a higher degree of variance in age (46.1 +/- 14.6 years). The immunosuppressive protocol was cyclosporine (CyA)-based regimen in related cases and tacrolimus-based in unrelated cases. RESULTS: We transplanted 16 left and 9 right kidneys from older donors. The mean cold ischemia time was 171 +/- 64 minutes with a second warm ischemia time of 24 +/- 6 minutes. Severe arteriosclerosis made vascular reconstruction by graft interposition necessary in two recipients. The acute rejection rate was 20%. Two patients (8%) required dialysis in the early postoperative course, whereas initial function was excellent in 22 patients (88%). The mean serum creatinine concentration after 12 months was 1.6 +/- 0.3 mg/dL (n = 24) and 2.0 +/- 0.7 mg/dL (n = 16) at 4 years. In comparison, the mean creatinine concentration after 4 years in donors under 60 years was 1.6 +/- 0.9 mg/dL. Our analysis showed no significant difference in long-term graft function comparing young versus old donors in the setting of living donor transplants. CONCLUSION: Using living donors older than 60 years for transplantation is a feasible and safe option. The difference in long-term creatinine between young and old donors was not significant.

Modelling gradient elution of bioactive multicomponent systems in non-linear ion-exchange chromatography.

A theoretical framework for the ion-exchange behaviour of bioactive substances in non-linear ion-exchange chromatogaphy is described. The aim of the study was the creation of a model basis to support a process design for production-scale ion-exchange chromatography. The theory can be applied to a whole variety of biological substances, such as amino acids, polysaccharides, peptides and proteins and either isocratic or gradient elution can be carried out. The influence of the eluent concentration on the ion-exchange as well as on the characteristic charge was considered. Experimental measurements showed a strong non-linear ion-exchange equilibrium with a transition from a Langmuir-type to a sigmoidal isotherm at higher eluent concentrations. Hereby, the compound binds to the surface though it is not ionic. Therefore, the model considered the possibility of ion-exchange as well as adsorption. A simplified distribution of the counter-ions based on the Gouy-Chapman theory with a discrete distribution of the counter-ions was used. The theory was extended by a selectivity in the double layer to allow specific adsorption. Calculations of adsorption-elution cycles showed, in agreement with the experimental observations, the development of non-linear elution profiles with a desorption fronting. As a result, the column loading and the eluent concentration were varied. The effect of contaminants, in this case sodium ions, was investigated and included in the model. Finally, the model was extended to multicomponent systems to investigate the effect of side components on the retention behaviour. The development of the characteristic elution profiles and the effect of the column loading on the separation are discussed. Calculated concentration profiles along the column at discrete time steps were used to reveal the influence of side components and the underlying separation mechanism. The simulations provided a new insight into the phenomena involved in biochromatography and make convenient design concepts at least doubtful as the separation is in this case mainly determined by the loading step and not by the choice of the elution gradient.

Desorption of a fixed-bed adsorber by ultrasound.

Adsorption processes offer the possibility to remove trace impurities of liquid process streams. An important step in adsorption processes is the regeneration of the adsorbent as it does not only affect the adsorption-desorption cycle but also the expenses of the following process steps. In this study, various desorption experiments of a loaded polymeric resin have been conducted. They were performed with cold and hot water as well as with ultrasound at different frequencies and intensities. The results indicate, that an important factor of ultrasound enhanced desorption is the temperature rise due to ultrasonication. But as regeneration is more effective at higher frequencies even though temperatures inside the column are lower, another ultrasonic effect occurs during ultrasonic desorption.

NH4NO3 extractable trace element contents of soil samples prepared for proficiency testing--a stability study.

In view of its intended use as a sample for proficiency testing or as a reference material the stability of the extractable trace element contents of a soil from an irrigation field was tested using the extraction with 1 mol/L ammonium nitrate solution according to DIN 19730. Therefore, changes of the extractability of sterilized and non sterilized soil samples stored at different temperatures were evaluated over a period of 18 months. Sets of bottles were kept at -20 degrees C, +4 degrees C, about +20 degrees C and +40 degrees C, respectively. The NH4NO3 extractable contents of Cd, Cr, Cu, Ni, Pb and Zn were determined immediately after bottling and then after 3, 6, 12 and 18 months with ICP-AES or ETAAS. Appropriate storage conditions are of utmost importance to prevent deterioration of soil samples prepared for the determination of NH4NO3 extractable trace element contents. Temperatures above +20 degrees C must be avoided. The observed changes in the extractability of the metals (especially for Cr and Cu) most likely could be related to thermal degradation of the organic matter of the soil. There is no need to sterilize dry soil samples, because microbiological activity in soils with a low moisture content appears to be negligible with regard to trace element mobilization.

Effect of the homogeneity of the column set on the performance of a simulated moving bed unit. I. Theory.

Although it is impossible to manufacture identical columns for use in a simulated moving bed (SMB) process, theoretical studies assume that all the columns in an SMB unit have identical characteristics. In practice, calculations in modeling and optimization studies are made with the average values of each column parameter set. In this report, the effects on SMB process performance caused by column-to-column fluctuations of the parameters are discussed. As a first step, we show how the differences in porosity of the columns may be taken into account with a revised set of separation conditions. Reductions in the purity of the extract and the raffinate streams are quantitatively related to the column-to-column fluctuations of the retention times of the two components arising from these porosity differences. For the sake of simplicity, the discussion first addresses the case of a four-column SMB operating under linear conditions. Then, the scope is extended to the cases of SMB units incorporating several columns in each section and to SMB units operating under nonlinear conditions.

Inhibition of neuronal nitric oxide synthase by 4-amino pteridine derivatives: structure-activity relationship of antagonists of (6R)-5,6,7,8-tetrahydrobiopterin cofactor.

The family of nitric oxide synthases (NOS) catalyzes the conversion of L-arginine to L-citrulline and nitric oxide (NO), an important cellular messenger molecule which has been implicated in the pathophysiology of septic shock and inflammatory and neurodegenerative disease states. NOS can be maximally activated by the ubiquitous cofactor, (6R)-5,6,7,8-tetrahydrobiopterin (H(4)Bip), and antagonists of H(4)Bip may be of therapeutic importance to inhibit pathologically high NO formation. The 4-amino substituted analogue of H(4)Bip was reported to be a potent NOS inhibitor. Therefore, we developed a series of novel 4-amino pteridine derivatives, anti-pterins, to pharmacologically target the neuronal isoform of nitric oxide synthase (NOS-I). To functionally characterize the pterin/anti-pterin interaction and establish a structure-activity relationship (SAR), we systematically altered the substituents in the 2-, 4-, 5-, 6-, and 7-position of the pteridine nucleus. Varying the substitution pattern in the 2-, 5-, and 7-position resulted in no significant inhibitory effect on enzyme activity. In contrast, bulky substituents in the 6-position, such as phenyl, markedly increased the inhibitory potency of the reduced 4-amino-5,6,7,8-tetrahydropteridines, possibly as a consequence of hydrophobic interactions within NOS-I. However, this was not the case for the aromatic 4-amino pteridines. Interestingly, chemical modification of the 4-amino substituent by dialkyl/diaralkylation together with 6-arylation of the aromatic 2,4-diamino pteridine resulted in potent and efficacious inhibitors of NOS-I, suggesting possible hydrophilic and hydrophobic interactions within NOS-I. This SAR agrees with (a) the recently published crystal structure of the oxygenase domain of the inducible NOS isoform (NOS-II) and (b) the comparative molecular field analysis of selected NOS-I inhibitors, which resulted in a 3D-QSAR model of the pterin binding site interactions. Further optimization should be possible when the full length structure of NOS-I becomes available.