Two-step postmortem angiography with a modified heart-lung machine: preliminary results

OBJECTIVE: The purpose of this study was to adapt and improve a minimally invasive two-step postmortem angiographic technique for use on human cadavers. Detailed mapping of the entire vascular system is almost impossible with conventional autopsy tools. The technique described should be valuable in the diagnosis of vascular abnormalities. MATERIALS AND METHODS: Postmortem perfusion with an oily liquid is established with a circulation machine. An oily contrast agent is introduced as a bolus injection, and radiographic imaging is performed. In this pilot study, the upper or lower extremities of four human cadavers were perfused. In two cases, the vascular system of a lower extremity was visualized with anterograde perfusion of the arteries. In the other two cases, in which the suspected cause of death was drug intoxication, the veins of an upper extremity were visualized with retrograde perfusion of the venous system. RESULTS: In each case, the vascular system was visualized up to the level of the small supplying and draining vessels. In three of the four cases, vascular abnormalities were found. In one instance, a venous injection mark engendered by the self-administration of drugs was rendered visible by exudation of the contrast agent. In the other two cases, occlusion of the arteries and veins was apparent. CONCLUSION: The method described is readily applicable to human cadavers. After establishment of postmortem perfusion with paraffin oil and injection of the oily contrast agent, the vascular system can be investigated in detail and vascular abnormalities rendered visible.

Hemozoin formation in malaria: a two-step process involving histidine-rich proteins and lipids

Major blood stage antimalarial drugs like chloroquine and artemisinin target the heme detoxification process of the malaria parasite. Hemozoin formation reactions in vitro using the Plasmodium falciparum histidine-rich protein-2 (Pfhrp-2), lipids, and auto-catalysis are slow and could not explain the speed of detoxification needed for parasite survival. Here, we show that malarial hemozoin formation is a coordinated two component process involving both lipids and histidine-rich proteins. Hemozoin formation efficiency in vitro is 1-2% with Pfhrp-2 and 0.25-0.5% with lipids. We added lipids after 9h in a 12h Pfhrp-2 mediated reaction that resulted in sixfold increase in hemozoin formation. However, a lipid mediated reaction in which Pfhrp-2 was added after 9h produced only twofold increase in hemozoin production compared to the reaction with Pfhrp-2 alone. Synthetic peptides corresponding to the Pfhrp-2 heme binding sequences, based on repeats of AHHAAD, neither alone nor in combination with lipids were able to generate hemozoin in vitro. These results indicate that hemozoin formation in malaria parasite involves both the lipids and the scaffolding proteins. Histidine-rich proteins might facilitate hemozoin formation by binding with a large number of heme molecules, and facilitating the dimer formation involving iron-carboxylate bond between two heme molecules, and lipids may then subsequently assist the mechanism of long chain formation, held together by hydrogen bonds or through extensive networking of hydrogen bonds.

THEODORE, a two-step heating system for the EC/OC determination of radiocarbon (14C) in the environment

Measurements of 14C in the organic carbon (OC) and elemental carbon (EC) fractions, respectively, of fine aerosol particles bear the potential to apportion anthropogenic and biogenic emission sources. For this purpose, the system THEODORE (two-step heating system for the EC/OC determination of radiocarbon in the environment) was developed. In this device, OC and EC are transformed into carbon dioxide in a stream of oxygen at 340 and 650 �C, respectively, and reduced to filamentous carbon. This is the target material for subsequent accelerator mass spectrometry (AMS) 14C measurements, which were performed on sub-milligram carbon samples at the PSI/ETH compact 500 kV AMS system. Quality assurance measurements of SRM 1649a, Urban Dust, yielded a fraction of modern fM in total carbon (TC) of 0.522 ±0.018 (n ¼ 5, 95% confidence level) in agreement with reported values. The results for OC and EC are 0.70± 0.05 (n ¼ 3) and 0.066 ± 0.020 (n ¼ 4), respectively.

Lake ice phenology from AVHRR data for European lakes: An automated two-step extraction method

Several lake ice phenology studies from satellite data have been undertaken. However, the availability of long-term lake freeze-thaw-cycles, required to understand this proxy for climate variability and change, is scarce for European lakes. Long time series from space observations are limited to few satellite sensors. Data of the Advanced Very High Resolution Radiometer (AVHRR) are used in account of their unique potential as they offer each day global coverage from the early 1980s expectedly until 2022. An automatic two-step extraction was developed, which makes use of near-infrared reflectance values and thermal infrared derived lake surface water temperatures to extract lake ice phenology dates. In contrast to other studies utilizing thermal infrared, the thresholds are derived from the data itself, making it unnecessary to define arbitrary or lake specific thresholds. Two lakes in the Baltic region and a steppe lake on the Austrian–Hungarian border were selected. The later one was used to test the applicability of the approach to another climatic region for the time period 1990 to 2012. A comparison of the extracted event dates with in situ data provided good agreements of about 10 d mean absolute error. The two-step extraction was found to be applicable for European lakes in different climate regions and could fill existing data gaps in future applications. The extension of the time series to the full AVHRR record length (early 1980 until today) with adequate length for trend estimations would be of interest to assess climate variability and change. Furthermore, the two-step extraction itself is not sensor-specific and could be applied to other sensors with equivalent near- and thermal infrared spectral bands.

A new technique to automatically quantify microstructures of fine grained carbonate mylonites: two-step etching combined with SEM imaging and image analysis

A two-step etching technique for fine-grained calcite mylonites using 0.37% hydrochloric and 0.1% acetic acid produces a topographic relief which reflects the grain boundary geometry. With this technique, calcite grain boundaries become more intensely dissolved than their grain interiors but second phase minerals like dolomite, quartz, feldspars, apatite, hematite and pyrite are not affected by the acid and therefore form topographic peaks. Based on digital backscatter electron images and element distribution maps acquired on a scanning electron microscope, the geometry of calcite and the second phase minerals can be automatically quantified using image analysis software. For research on fine-grained carbonate rocks (e.g. dolomite calcite mixtures), this low-cost approach is an attractive alternative to the generation of manual grain boundary maps based on photographs from ultra-thin sections or orientation contrast images.

A fluorescently labeled dendronized polymer-enzyme conjugate carrying multiple copies of two different types of active enzymes

A hybrid structure of a synthetic dendronized polymer, two different types of enzymes (superoxide dismutase and horseradish peroxidase), and a fluorescent dye (fluorescein) was synthesized. Thereby, a single polymer chain carried multiple copies of the two enzymes and the fluorescein. The entire attachment chemistry is based on UV/vis-quantifiable bis-aryl hydrazone bond formation that allows direct quantification of bound molecules: 60 superoxide dismutase, 120 horseradish peroxidase, and 20 fluorescein molecules on an average polymer chain of 2000 repeating units. To obtain other enzyme ratios the experimental conditions were altered accordingly. Moreover, it could be shown that both enzymes remained fully active and catalyzed a two-step cascade reaction.

Two-dimensional assembly and local redox-activity of molecular hybrid structures in an electrochemical environment

The self-assembly and redox-properties of two viologen derivatives, N-hexyl-N-(6-thiohexyl)-4,4-bipyridinium bromide (HS-6V6-H) and N,N-bis(6-thiohexyl)-4,4-bipyridinium bromide (HS-6V6-SH), immobilized on Au(111)-(1x1) macro-electrodes were investigated by cyclic voltammetry, surface enhanced infrared spectroscopy (SEIRAS) and in situ scanning tunneling microscopy (STM). Depending on the assembly conditions one could distinguish three different types of adlayers for both viologens: a low coverage disordered and an ordered striped phase of flat oriented molecules as well as a high coverage monolayer composed of tilted viologen moieties. Both molecules, HS-6V6-H and HS-6V6-SH, were successfully immobilized on Au(poly) nano-electrodes, which gave a well-defined redox-response in the lower pA–current range. An in situ STM configuration was employed to explore electron transport properties of single molecule junctions Au(T)|HS-6V6-SH(HS-6V6-H)|Au(S). The observed sigmoidal potential dependence, measured at variable substrate potential ES and at constant bias voltage (ET–ES), was attributed to electronic structure changes of the viologen moiety during the one-electron reduction/re-oxidation process V2+ V+. Tunneling experiments in asymmetric, STM-based junctions Au(T)-S-6V6-H|Au(S) revealed current (iT)–voltage (ET) curves with a maximum located at the equilibrium potential of the redox-process V2+ V+. The experimental iT–ET characteristics of the HS-6V6-H–modified tunneling junction were tentatively attributed to a sequential two-step electron transfer mechanism.

Surface Roughness and Microhardness of Two Recent CAD/CAM-Materials After Storage

Objectives: To investigate surface roughness and microhardness of two recent resin-ceramic materials for computer-aided design/computer-aided manufacturing (CAD/CAM) after polishing with three polishing systems. Surface roughness and microhardness were measured immediately after polishing and after six months storage including monthly artificial toothbrushing. Methods: Sixty specimens of Lava Ultimate (3M ESPE) and 60 specimens of VITA ENAMIC (VITA Zahnfabrik) were roughened in a standardized manner and polished with one of three polishing systems (n=20/group): Sof-Lex XT discs (SOFLEX; three-step (medium-superfine); 3M ESPE), VITA Polishing Set Clinical (VITA; two-step; VITA Zahnfabrik), or KENDA Unicus (KENDA; one-step; KENDA Dental). Surface roughness (Ra; μm) was measured with a profilometer and microhardness (Vickers; VHN) with a surface hardness indentation device. Ra and VHN were measured immediately after polishing and after six months storage (tap water, 37°C) including monthly artificial toothbrushing (500 cycles/month, toothpaste RDA ~70). Ra- and VHN-values were analysed with nonparametric ANOVA followed by Wilcoxon rank sum tests (α=0.05). Results: For Lava Ultimate, Ra (mean [standard deviation] before/after storage) remained the same when polished with SOFLEX (0.18 [0.09]/0.19 [0.10]; p=0.18), increased significantly with VITA (1.10 [0.44]/1.27 [0.39]; p=0.0001), and decreased significantly with KENDA (0.35 [0.07]/0.33 [0.08]; p=0.03). VHN (mean [standard deviation] before/after storage) decreased significantly regardless of polishing system (SOFLEX: 134.1 [5.6]/116.4 [3.6], VITA: 138.2 [10.5]/115.4 [5.9], KENDA: 135.1 [6.2]/116.7 [6.3]; all p<0.0001). For VITA ENAMIC, Ra (mean [standard deviation] before/after storage) increased significantly when polished with SOFLEX (0.37 [0.18]/0.41 [0.14]; p=0.01) and remained the same with VITA (1.32 [0.37]/1.31 [0.40]; p=0.58) and with KENDA (0.81 [0.35]/0.78 [0.32]; p=0.21). VHN (mean [standard deviation] before/after storage) remained the same regardless of polishing system (SOFLEX: 284.9 [24.6]/282.4 [31.8], VITA: 284.6 [28.5]/276.4 [25.8], KENDA: 292.6 [26.9]/282.9 [24.3]; p=0.42-1.00). Conclusion: Surface roughness and microhardness of Lava Ultimate was more affected by storage and artificial toothbrushing than was VITA ENAMIC.

Congenital cervical kyphosis in two young sighthounds

Introduction: Cervical vertebral (C) malformation is rarely reported in large breed dogs. Congenital cervical kyphosis (CCK) may result from defects of vertebral segmentation, failure of formation or both. This report describes two cases of C3-C4 CCK in young sighthounds, treated surgically. Case description: An 18-month-old female Deerhound and a six-week-old female Borzoi dog were presented because of the complaints of reluctance to exercise and signs of of neck pain. Both dogs were neurologically normal. Diagnostic imaging revealed C3-C4 deformity, moderate kyphosis, and spinal canal stenosis associated with chronic spinal cord pressure atrophy. Both dogs underwent surgical treatment. Results: A staged two-step surgery starting with dorsal decompression was elected in the Deerhound. After the first surgical procedure, the dog developed focal myelomalacia and phrenic nerve paralysis and was euthanatized. A ventral distraction-fusion technique with two locking plates was performed in the Borzoi. This patient recovered uneventfully and long-term follow-up computed tomography revealed complete spondylodesis. Clinical significance: Until now, CCK has only been described in sighthounds. Congenital cervical kyphosis might be considered a differential diagnosis in these breeds that are presented with signs of cervical pain. Ventral realignment-fusion and bone grafting may be considered for surgical treatment, although the earliest age at which this procedure can and should be performed remains unclear.

Next-state comfort in learning a vertical stick transportation sequence

Over the last decade, the end-state comfort effect (e.g., Rosenbaum et al., 2006) has received a considerable amount of attention. However, some of the underlying mechanisms are still to be investigated, amongst others, how sequential planning affects end-state comfort and how this effect develops over learning. In a two-step sequencing task, e.g., postural comfort can be planned on the intermediate position (next state) or on the actual end position (final state). It might be hypothesized that, in initial acquisition, next state’s comfort is crucial for action planning but that, in the course of learning, final state’s comfort is taken more and more into account. To test this hypothesis, a variant of Rosenbaum’s vertical stick transportation task was used. Participants (N = 16, right-handed) received extensive practice on a two-step transportation task (10,000 trials over 12 sessions). From the initial position on the middle stair of a staircase in front of the participant, the stick had to be transported either 20 cm upwards and then 40 cm downwards or 20 cm downwards and then 40 cm upwards (N = 8 per subgroup). Participants were supposed to produce fluid movements without changing grasp. In the pre- and posttest, participants were tested on both two-step sequencing tasks as well as on 20 cm single-step upwards and downwards movements (10 trials per condition). For the test trials, grasp height was calculated kinematographically. In the pretest, large end/next/final-state comfort effects for single-step transportation tasks and large next-state comfort effects for sequenced tasks were found. However, no change in grasp height from pre- to posttest could be revealed. Results show that, in vertical stick transportation sequences, the final state is not taken into account when planning grasp height. Instead, action planning seems to be solely based on aspects of the next action goal that is to be reached.

Retrospective analysis of stored dried blood spots from children with cystic fibrosis and matched controls to assess the performance of a proposed newborn screening protocol in Switzerland

Newborn screening (NBS) for Cystic Fibrosis (CF) has been introduced in many countries, but there is no ideal protocol suitable for all countries. This retrospective study was conducted to evaluate whether the planned two step CF NBS with immunoreactive trypsinogen (IRT) and 7 CFTR mutations would have detected all clinically diagnosed children with CF in Switzerland.

Conformationally Controlled Electron Delocalization in n-Type Rods: Synthesis, Structure, and Optical, Electrochemical, and Spectroelectrochemical Properties of Dicyanocyclophanes

A series of dicyanobiphenyl-cyclophanes 1-6 with various pi-backbone conformations and characteristic n-type semiconductor properties is presented. Their synthesis, optical, structural, electrochemical, spectroelectrochemical, and packing properties are investigated. The X-ray crystal structures of all n-type rods allow the systematic correlation of structural features with physical properties. In addition, the results are supported by quantum mechanical calculations based on density functional theory. A two-step reduction process is observed for all n-type rods, in which the first step is reversible. The potential gap between the reduction processes depends linearly on the cos(2) value of the torsion angle phi between the pi-systems. Similarly, optical absorption spectroscopy shows that the vertical excitation energy of the conjugation band correlates with the cos(2) value of the torsion angle phi. These correlations demonstrate that the fixed intramolecular torsion angle phi is the dominant factor determining the extent of electron delocalization in these model compounds, and that the angle phi measured in the solid-state structure is a good proxy for the molecular conformation in solution. Spectroelectrochemical investigations demonstrate that conformational rigidity is maintained even in the radical anion form. In particular, the absorption bands corresponding to the SOMO-LUMO+i transitions are shifted bathochromically, whereas the absorption bands corresponding to the HOMO-SOMO transition are shifted hypsochromically with increasing torsion angle phi.