Generation of co-culture spheroids as vascularisation units for bone tissue engineering.

Cell spheroids represent attractive building units for bone tissue engineering, because they provide a three-dimensional environment with intensive direct cell-cell contacts. Moreover, they allow for co-culture of both osteoblasts and vessel-forming cells, which may markedly increase their survival and vascularisation after transplantation. To test this hypothesis, we generated co-culture spheroids by aggregating different combinations of primary human osteoblasts (HOB), human dermal microvascular endothelial cells (HDMEC) and normal human dermal fibroblasts (NHDF) using the liquid overlay technique. Mono-culture spheroids consisting either of HOB or HDMEC served as controls. After in vitro characterisation, the different spheroids were transplanted into dorsal skinfold chambers of CD1 nu/nu mice to study in vivo their viability and vascularisation over a 2-week observation period by means of repetitive intravital fluorescence microscopy and immunohistochemistry. In vitro, co-culture spheroids containing HDMEC rapidly formed dense tubular vessel-like networks within 72 h and exhibited a significantly decreased rate of apoptotic cell death when compared to mono-culture HDMEC spheroids. After transplantation, these networks interconnected to the host microvasculature by external inosculation. Of interest, this process was most pronounced in HOB-HDMEC spheroids and could not further be improved by the addition of NHDF. Accordingly, HOB-HDMEC spheroids were larger when compared to the other spheroid types. These findings indicate that HOB-HDMEC spheroids exhibit excellent properties to preserve viability and to promote proliferation and vascularisation. Therefore, they may be used as functional vascularisation units in bone tissue engineering for the seeding of scaffolds or for the vitalisation of non-healing large bone defects.

Interferometry with Bose-Einstein condensates in microgravity.

Atom interferometers covering macroscopic domains of space-time are a spectacular manifestation of the wave nature of matter. Because of their unique coherence properties, Bose-Einstein condensates are ideal sources for an atom interferometer in extended free fall. In this Letter we report on the realization of an asymmetric Mach-Zehnder interferometer operated with a Bose-Einstein condensate in microgravity. The resulting interference pattern is similar to the one in the far field of a double slit and shows a linear scaling with the time the wave packets expand. We employ delta-kick cooling in order to enhance the signal and extend our atom interferometer. Our experiments demonstrate the high potential of interferometers operated with quantum gases for probing the fundamental concepts of quantum mechanics and general relativity.

Bose-Einstein condensation in microgravity.

Albert Einstein's insight that it is impossible to distinguish a local experiment in a "freely falling elevator" from one in free space led to the development of the theory of general relativity. The wave nature of matter manifests itself in a striking way in Bose-Einstein condensates, where millions of atoms lose their identity and can be described by a single macroscopic wave function. We combine these two topics and report the preparation and observation of a Bose-Einstein condensate during free fall in a 146-meter-tall evacuated drop tower. During the expansion over 1 second, the atoms form a giant coherent matter wave that is delocalized on a millimeter scale, which represents a promising source for matter-wave interferometry to test the universality of free fall with quantum matter.

Wave refraction in negative-index media: always positive and very inhomogeneous.

We present the first treatment of the refraction of physical electromagnetic waves in newly developed negative index media (NIM), also known as left-handed media (LHM). The NIM dispersion relation implies that group fronts refract positively even when phase fronts refract negatively. This difference results in rapidly dispersing, very inhomogeneous waves. In fact, causality and finite signal speed always prevent negative wave signal (not phase) refraction. Earlier interpretations of phase refraction as "negative light refraction" and "light focusing by plane slabs" are therefore incorrect, and published NIM experiments can be explained without invoking negative signal refraction.

Resonance superfluidity in a quantum degenerate Fermi gas.

We consider the superfluid phase transition that arises when a Feshbach resonance pairing occurs in a dilute Fermi gas. We apply our theory to consider a specific resonance in potassium ((40)K), and find that for achievable experimental conditions, the transition to a superfluid phase is possible at the high critical temperature of about 0.5T(F). Observation of superfluidity in this regime would provide the opportunity to experimentally study the crossover from the superfluid phase of weakly coupled fermions to the Bose-Einstein condensation of strongly bound composite bosons.

Comparison of different schemes to treat long-range electrostatic interactions in molecular dynamics simulations of a protein crystal.

Eight molecular dynamics simulations of a ubiquitin crystal unit cell were performed to investigate the effect of different schemes to treat the long-range electrostatic interactions as well as the need to include counter ions. A crystal system was chosen as the test system, because the higher charge density compared with a protein in solution makes it more sensitive to the way of treating the electrostatic interactions. Three different schemes of treating the long-range interactions were compared: straight cutoff, reaction-field approximation, and a lattice-sum method (P3M). For each of these schemes, two simulations were performed, one with and one without the counter ions. Two additional simulations with a reaction-field force and different initial placements of the counter ions were performed to examine the effect of the initial positions of the ions. The inclusion of long-range electrostatic interactions using either a reaction-field or a lattice-sum method proved to be necessary for the simulation of crystals. These two schemes did not differ much in their ability to reproduce the crystallographic structure. The inclusion of counter ions, on the other hand, seems not necessary for obtaining a stable simulation. The initial positions of the ions have a visible but small effect on the simulation.

Formation of pairing fields in resonantly coupled atomic and molecular bose-einstein condensates.

We show that pair correlations may play an important role in the dynamical properties of a Bose-Einstein condensed gas composed of an atomic field resonantly coupled with a condensed field of molecular dimers. Specifically, pair correlations in this system can dramatically modify the coherent and incoherent transfers between the atomic and molecular fields.

Viscosity dependence of protein dynamics.

The influence of solvent viscosity on protein dynamics was investigated with molecular dynamics simulations of factor Xa in two solvents differing only in viscosity, by a factor of 10. We obtained this viscosity change by changing the masses of the solvent atoms by a factor of 100. Equilibrium properties of the protein, that is, the average structure, its fluctuations, and the secondary structure, show no significant dependence on the solvent viscosity. The dynamic properties of the protein, that is, the atom-positional correlation times and torsional angle transitions, however, depend on the solvent viscosity. The protein appears to be much more mobile in the solvent of lower viscosity. It feels the influence of the solvent not only on the surface but even in its core. With increasing solvent viscosity, the positional relaxation times of atoms in the protein core increase as much as those of atoms on the protein surface, and the relative increase in the core is even larger than on the surface.

On the temperature and pressure dependence of a range of properties of a type of water model commonly used in high-temperature protein unfolding simulations.

Molecular dynamics simulations of protein folding and unfolding are often carried out at temperatures (400-600 K) that are much higher than physiological or room temperature to speed up the (un)folding process. Use of such high temperatures changes both the protein and solvent properties considerably, compared to physiological or room temperature. Water models designed for use in conjunction with biomolecules, such as the simple point charge (SPC) model, have generally been calibrated at room temperature and pressure. To determine the distortive effect of high simulation temperatures on the behavior of such "room temperature" water models, the structural, dynamic, and thermodynamic properties of the much-used SPC water model are investigated in the temperature range from 300 to 500 K. Both constant pressure and constant volume conditions, as used in protein simulations, were analyzed. We found that all properties analyzed change markedly with increasing temperature, but no phase transition in this temperature range was observed.

The arterial blood supply of the human patella. Its clinical importance for the operating technique in vascularized knee joint transplantations.

The architecture of the arteries supplying the patellar rete was examined in 14 anatomic specimens in order to develop an optimized operating technique for knee joint transplantation. The specimens were fixed in Jores Solution and exarticulated from the hip joint. The lower limbs were injected with Berliner-Blau-Gelatin, and the arteries were dissected macroscopically. Five to six main arteries entered the patellar rete at 1, 3, 5, 7 and 11 o'clock forming an arterial circle. These arteries were the same main arteries which supply the distal end of the femur and the proximal part of the tibia. From an anatomic perspective, they provide the complete arterial blood supply to a whole knee joint being transplanted including the patella. Based on these anatomic results, we transplanted two allogenic vascularized human knee joints preserving the patella, the capsule, and the patellar ligament. Up to six months after surgery we demonstrated the perfusion and viability of all three transplanted bones, particularly the patella, by 99mTc DPD scintigraphy. We compared these findings with knee joint arthroscopy and with histologic results from biopsies taken from the patella. The postoperative examinations clearly indicated the viability of the transplanted patella employing this new operating technique. The results of the entire study demonstrate that it is technically feasible to transplant a whole knee joint which remains clinically viable.

[201Tl myocardial SPECT. First experiences with a simultaneous transmission-emission acquisition protocol for patient-specific attenuated correction]. / 201Ti-Myokard-SPECT. Erste Erfahrungen mit einem simultanen Transmission-Emission-Akquisitionsprotokoll zur patientenspezifischen Abschwächungskorrektur.

AIM: In this study our first clinical experiences with simultaneous transmission and emission acquisition in 201 TI myocardial SPECT (T/E-SPECT) are discussed. METHODS: The non-uniform attenuation (AK) was carried out with a triple-head camera (PRISM 3000, Picker Inc.) correction equipped with fanbeam collimators. A line source of 750 MBq 99mTc was used to construct the transmission profile. Prior to investigation patients got 80-120 MBq 201TI-chloride intravenously injected. RESULTS: The study comprises the evaluation of 40 patients, derived from the clinical routine. The investigation followed an usual one day protocol. Our results using T/E-SPECT reveal an almost equilibrated activity distribution between anterior and posterior myocardial wall. CONCLUSION: For this reason it is to be expected that T/E-SPECT provides more reliable information about the posterior myocardial wall, than the usual SPECT technique without attenuation correction.

Diagnostic angioscintigraphic evaluation of malignant hepatic tumors before catheter embolization: determination of shunt, flow distribution, and reflux.

PURPOSE: The aim of this study was to evaluate quantitatively arteriovenous shunts in malignant liver tumors by injection of 99mTc macroaggregates of albumin (MAA) into the tumor-feeding artery after selective catheterization. METHODS: In 40 patients with malignant liver tumors (33 hepatocellular carcinomas and 7 metastases of colorectal cancer), a mean dose of 200 MBq 99mTC MAA was injected arterially during angiography. The embolized area and the lungs were then visualized using a gamma camera. A dedicated computer program calculated pulmonary shunt rates. RESULTS: The majority of patients (n = 30) with hepatocellular carcinoma showed small shunts varying from 0 to 15%; only 3 of these patients had shunts ranging from 18% to 37%. In patients with colorectal carcinoma metastases (n = 7) the shunt varied from 0 to 3% (2 +/- 1%), probably due to a physiological shunt in normal liver tissue in the embolized area. Importantly, the degree of shunt found bore no correlation to the tumor volume or to the pattern of vascularity on angiography. CONCLUSION: Diagnostic angioscintigraphy is a useful tool for pretherapeutic evaluation of the capacity of an individual tumor to retain particles and to measure extratumoral shunting; these are essential for therapy planning, as they can help to increase the safety and effectiveness of embolization.

Communicating behavioral views and techniques to practicing clinicians.

The chasm between behavioral and nonbehavioral views of clinical phenomena results in communication difficulties in both directions. We distinguish five situations based on the nature and degree of differences in understanding within the two language systems. We recommend different communication strategies for behavioral scientists, depending upon which situation best applies.

Diode-laser noise spectroscopy of rubidium.

We report on spectra obtained by measuring the laser intensity noise after a broad-bandwidth diode-laser beam passes through a rubidium vapor cell. The atomic resonance converts laser frequency fluctuations into intensity fluctuations. We compare our experimental spectra with numerically calculated spectra based on a phase-diffusion model of the laser field and find good agreement.

Mechanisms for laser-induced functional damage to silicon charge-coupled imaging sensors.

We measured the functional degradation of silicon CCD photodetector arrays when subjected to Nd:YAG laser irradiation at 1.06 µm by 10-ns pulses. Operational tests such as dark leakage, point-spread function, and modulation transfer function were developed for testing individual pixels and applied to the testing of locally laser-damaged CCD arrays. Testing revealed that the primary failure mechanism was the spreading of the point-spread function in the direction of clocked charge motion that resulted from a decreased depth of potential wells within the laser-damaged spot. Lesser degradation was observed at pixels near the damaged spot that were served by clock lines that traversed the damage spot. This damage behavior was correlated with decreased breakdown voltage and increased leakage current between adjacent clock lines. Subsequent morphological and TEM examination of similarly constructed chips indicated that laser heating of the polysilicon clock lines led to degradation of the adjacent isolation oxide between clock lines. Functional damage occurred at locations where two clock lines were very close to each other, and the thin oxide layer separating them was subjected to high temperatures that resulted from melting of the neighboring polysilicon.