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Biomedical subjects

P Walther

Publications and source records attributed to P Walther.

At least 19 recordsLinked to original sources

Timing and patterns of recurrences and deaths from prostate cancer following adjuvant pelvic radiotherapy for pathologic stage T3/4 adenocarcinoma of the prostate.

To determine the timing and patterns of late recurrence after radical prostatectomy (RP) alone or RP plus adjuvant radiotherapy (RT). Between 1970 and 1983, 159 patients underwent RP for newly diagnosed adenocarcinoma of the prostate and were found to have positive surgical margins, extracapsular extension and/or seminal vesicle invasion. Of these, 46 received adjuvant RT and 113 did not. The RT group generally received 45-50 Gy to the whole pelvis, then a boost to the prostate bed (total dose of 55-65 Gy). In the RP group, 62% received neoadjuvant/adjuvant androgen deprivation vs 17% in the RT group. Patients were analyzed with respect to timing and patterns of failure. Only one patient was lost to follow-up. The median follow-up for surviving patients was nearly 20 years. The median time to failure in the surgery group was 7.5 vs 14.7 years in the RT group (P=0.1). Late recurrences were less common in the surgery group than the RT group (9 and 1% at 10 and 15 years, respectively vs 17 and 9%). In contrast to recurrences, nearly half of deaths from prostate cancer occurred more than 10 years after treatment. Deaths from prostate cancer represented 55% of all deaths in these patients. Recurrences beyond 10 years after RP in this group of patients were relatively uncommon. Despite its long natural history, death from prostate cancer was the most common cause of mortality in this population with locally advanced tumors, reflecting the need for more effective therapy.

Adenocarcinoma↗

Experimental one-way quantum computing.

Standard quantum computation is based on sequences of unitary quantum logic gates that process qubits. The one-way quantum computer proposed by Raussendorf and Briegel is entirely different. It has changed our understanding of the requirements for quantum computation and more generally how we think about quantum physics. This new model requires qubits to be initialized in a highly entangled cluster state. From this point, the quantum computation proceeds by a sequence of single-qubit measurements with classical feedforward of their outcomes. Because of the essential role of measurement, a one-way quantum computer is irreversible. In the one-way quantum computer, the order and choices of measurements determine the algorithm computed. We have experimentally realized four-qubit cluster states encoded into the polarization state of four photons. We characterize the quantum state fully by implementing experimental four-qubit quantum state tomography. Using this cluster state, we demonstrate the feasibility of one-way quantum computing through a universal set of one- and two-qubit operations. Finally, our implementation of Grover's search algorithm demonstrates that one-way quantum computation is ideally suited for such tasks.

Journal Article↗

Full characterization of a three-photon Greenberger-Horne-Zeilinger state using quantum state tomography.

We have performed the first experimental tomographic reconstruction of a three-photon polarization state. Quantum state tomography is a powerful tool for fully describing the density matrix of a quantum system. We measured 64 three-photon polarization correlations and used a "maximum-likelihood" reconstruction method to reconstruct the Greenberger-Horne-Zeilinger state. The entanglement class has been characterized using an entanglement witness operator and the maximum predicted values for the Mermin inequality were extracted.

Journal Article↗

Quantum nonlocality obtained from local states by entanglement purification.

We have applied an entanglement purification protocol to produce a single entangled pair of photons capable of violating a Clauser-Horne-Shimony-Holt Bell inequality from two pairs that individually could not. The initial poorly entangled photons were created by a controllable decoherence that introduced complex errors. All of the states were reconstructed using quantum state tomography which allowed for a quantitative description of the improvement of the state after purification.

Journal Article↗

[Does the orientation of the IOL in the eye play a role? Imaging quality of foldable intraocular lenses measured utilizing point spread function (PSF-measurement) and scanning electron microscopy cross-sectional images].

BACKGROUND: Foldable intraocular lenses (IOLs) are established in ophthalmic surgery and increasingly more complex geometries are available. This study investigates different designs of the optics of the IOL as well as the imaging quality of these optics utilizing electron microscopic photographs and mathematical approximations. METHOD: Included in this study were two different lens powers of common IOLs (+ 13 D and + 26 D). All studied IOLs were processed depending upon their water content: Aqueous IOLs with up to 25 % water content were fixed with 2.5 % glutaraldehyde in 0.1 M phosphate buffer (mPP), and then contrasted with 2 % OsO (4) and processed in an alcohol series of 30 - 100 % propanol. Subsequently, the IOLs were transferred into EPON (epoxy resin) and polymerization took place at 60 degrees C. After hardening, the samples were cut out with a leaves saw and divided into halves. Afterwards these were polished with glass and diamond knifes at the ultramicrotom (Ultracut, Rückert-Jung). After coal vaporization measurement took place in the scanning electron microscope (DSM 962, Zeiss). In the case of dry stored IOLs, the samples were sputtered with Au-Pd 20 nm and afterwards measurement was performed as described above. The measurement of the optical imaging quality was performed with correctly and inversely oriented IOLs according to the standard EN ISO 11979 -- 2 by means of a measuring system which works on the basis of the point spread function (PSF) and/or the modulation transfer function (MTF). From this the Strehl ratio was computed in each case. RESULTS: Utilizing a correlation of the electron microscopic investigation of the optics of different IOLs with their imaging quality (Strehl ratio), we were able to show that the optical design of the IOL has substantial influence on the imaging quality of an intraocular lens. In particular, with non-symmetrical designs of the optics (e. g., AR40 e; AMO) the orientation of an IOL in the eye is crucial; i. e., the Strehl ratio increased thereby from 0.68 to 0.87. CONCLUSIONS: The design of asymmetrical optics is crucial and much attention should be paid to the fact that the imaging quality can substantially be affected. A correct orientation of the lens within the eye is very important with these asymmetrical optics.

Algorithms↗

Quantitative analysis of keratin filament networks in scanning electron microscopy images of cancer cells.

The keratin filament network is an important part of the cytoskeleton. It is involved in the regulation of shape and viscoelasticity of epithelial cells. The morphology of keratin networks depends on post-translational modifications of keratin monomers. In-vitro studies indicated that network characteristics, such as filament crosslink density, determines the biophysical properties of the filament network. This report presents a quantitative method for the morphological analysis of keratin filament networks. Visualization of filaments was based on prefixation extraction of epithelial cells and scanning electron microscopy (SEM). SEM images were processed by a skeletonization algorithm to obtain a graph structure that represents individual filaments as well as their connections. This method was applied to investigate the effects of transforming growth factor alpha (TGFalpha) on the morphology of keratin networks in pancreatic cancer cells. TGFalpha contributes to pancreatic cancer progression and activates signalling pathways phosphorylating keratin monomers. Using this new method, a significant alteration to the keratin network morphology could be detected in response to TGFalpha.

Cell Transformation, Neoplastic↗

Influence of chemical and freezing fixation methods in the freeze-fracture of stratum corneum.

A comparison between two fixation techniques for freeze-fracture was established. Stratum corneum (SC) samples from pig epidermis were fixed using high-pressure freezing (HPF) and using plunging in propane freezing; the latter after chemical fixation. Then, frozen samples were freeze-fractured, coated with platinum-carbon, and visualized using a high-resolution low-temperature scanning electron microscope and a transmission electron microscope. Our results indicate that the plane of freeze-fracture was different depending on the fixation and freezing methodology used. In the samples frozen by HPF without chemical fixation, the fracture plane laid mainly between the lipid lamellae. However, when chemical fixation and plunging in propane freezing was used, the fracture plane did not show preference to a specific way. Plunging in propane freezing of chemically fixed samples, on the other hand, provides a more homogeneous fracture behaviour. Thus, depending on the methodology used, we can favour a visualization of either lipid or protein domains of the SC. These results could be very useful in future ultrastructural studies in order to facilitate the microscopic visualization and interpretation of the complex images such as those of SC and even of other samples in which different domains coexist.

Animals↗

Recent progress in freeze-fracturing of high-pressure frozen samples.

Pancreatic tissue, bacteria and lipid vesicles were high-pressure frozen and freeze-fractured. In addition to the normal holder, a new type of high-pressure freezing holder was used that is particularly suitable for suspensions. This holder can take up an EM grid that has been dipped in the suspension and clamped in between two low-mass copper platelets, as used for propane-jet freezing. Both the standard and the new suspension holder allowed us to make cryo-fractures without visible ice crystal damage. High-pressure frozen rat pancreas tissue samples were cryo-fractured and cryo-sectioned with a new type diamond knife in the microtome of a freeze-etching device. The bulk fracture faces and blockfaces were investigated in the frozen-hydrated state by use of a cryo-stage in an in-lens SEM. Additional structures can be made visible by controlled sublimation of ice ('etching'), leading to a better understanding of the three-dimensional organization of organelles, such as the endoplasmic reticulum. With this approach, relevant biological structures can be investigated with a few nanometre resolution in a near life-like state, preventing the artefacts associated with conventional fixation techniques.

Animals↗

[Intensive care of delirium syndromes].

Delirium is mental dysfunctions occurring as impaired attentional and memory systems with disturbances of consciousness, affectivity, psychomotor activity and sleep patterns. Numerous factors and underlying diseases may be responsible for these non-specific symptoms. Therefore, a thorough evaluation of preadmission history and current clinical status, supplemented by laboratory and extended technical diagnostic procedures, are always required. If delirium occurs in connection with emergency admission to hospital, an organic disease can most regularly be found. Due to its rapid time of onset and minor side-effects, the intravenous injection of 2.0 g gamma-hydroxybutyric acid is preferred for sedation of extremely agitated patients. Neuroleptic drugs are indicated in psychiatric patients. A central anticholinergic syndrome in the early postoperative period causative of the symptoms of delirium may respond to intravenous injection of physostigmine. Most of the time, however, these acute disturbances of brain function are best treated by correction of homeostatic imbalances, restoration of cardiovascular and respiratory stability and alleviation of pain. Postoperative delirium occurring two or more days later is frequently due to respiratory distress, followed by sepsis, alcohol withdrawal and many other causes including heart failure, exsiccosis and side-effects of drugs. In intensive care patients, delirium may be caused, for example, by withdrawal (alcohol, opioids, benzodiazepines), the onset of sepsis (often venous catheter related), side-effects of drugs, problems of communication, sleep deprivation and others. Treatment should focus on finding the right approach. Personal care should be intensified and include help from family members. Most problems arise from agitated, non-cooperative patients. Treatment with clonidine, gamma-hydroxybutyric acid or neuroleptic drugs like perazin and haloperidol may be required to reduce agitation and the activation of sympathetic influence.

Critical Care↗

Freeze substitution of high-pressure frozen samples: the visibility of biological membranes is improved when the substitution medium contains water.

Biological membranes are often poorly visible with the electron microscope after high-pressure freezing and freeze-substitution. The water content of the sample and of the substitution medium is one factor among others that strongly influences membrane visibility. In order to investigate this effect, high-pressure frozen yeast cells, rat-pancreas tissue and arthropod tissue were freeze-substituted with and without adding water to the substitution medium. The visibility of the biological membranes was generally improved if the substitution medium contained 1-5% water. The effect was especially pronounced in yeast cells, where membrane visibility was poor after freeze-substitution with water-free medium but good after addition of 5% water to the substitution medium.

Animals↗

Liposomes as protective agents of stratum corneum against octyl glucoside: a study based on high-resolution, low-temperature scanning electron microscopy.

The ability of phosphatidylcholine (PC) liposomes to protect pig stratum corneum (SC) against the action of the nonionic surfactant octyl glucoside (OG) was investigated "in vitro" using double-layer coating for high-resolution, low-temperature scanning electron microscopy. This technique has been useful in preventing drying artifacts in the study of biological materials. The treatment of SC with OG led to a perturbation mainly in the corneocytes. However, the incubation of the tissue with liposomes prior to the OG treatment resulted in a progressive decrease in these perturbations and, consequently, in the progressive protection of the SC against the action of the surfactant.

Animals↗

Calcium stores in differentiated Dictyostelium discoideum: prespore cells sequester calcium more efficiently than prestalk cells.

Dictyostelium discoideum pseudoplasmodia exhibit a gradient of the cytosolic free Ca2+-concentration ([Ca2+]i) along their anterior-posterior axis involved in cell-type specific differentiation. [Ca2+]i is high in prestalk and low in prespore cells. We determined the content and localization of calcium and other elements in cryosectioned cells of pseudoplasmodia and fruiting bodies by X-ray microanalysis. Granular stores rich in Ca, Mg and P were identified. Average Ca was higher in prespore than prestalk granules (225vs 111 mmol/kg dry weight). Total Ca stored in granules was also higher in prespore than prestalk cells. The amount of P and S in granules differed between the two cell types indicating different store composition. In spores mean granular Ca was 120 mmol/kg dry weight. Stalk cells had smaller granules with 360 mmol Ca/kg dry weight. Complementary to microanalysis, vesicular Ca2+-fluxes were studied in fractionated cell homogenates. The rate of Ca2+-uptake was higher in pellet fractions of prespore than prestalk amoebae (4.7 vs 3.4 nmol/min x mg). Ca2+-release was greater in supernatant fractions from prestalk than prespore cells (16.5vs 7.7 nmol/10(8)cells). In summary, prestalk and prespore cells possess qualitatively different, high-capacity stores containing distinct amounts of Ca and probably being involved in regulation of the anterior-posterior [Ca2+]i-gradient.

Animals↗

Immunohistochemical demonstration of hyaluronan and its possible involvement in axolotl neural crest cell migration.

Hyaluronan (HA), an extracellular matrix component, is involved mainly in the control of cell proliferation, neural crest and tumor cell migration, and wound repair. We investigated the effect of hyaluronan on neural crest (NC) cell migration and its ultrastructural localization in dark (wild-type) and white mutant embryos of the Mexican axolotl (Ambystoma mexicanum, Amphibia). The axolotl system is an accepted model for studying mechanisms of NC cell migration. Using a biotinylated hyaluronan binding protein (HABP), major extracellular matrix (ECM) spaces, including those of NC cell migration, reacted equally positive on cryosections through dark and white embryos. Since neural crest-derived pigment cells migrate only in subepidermal spaces of dark embryos, HA does not seem to influence crest cell migration in vivo. However, when tested on different alternating substrates in vitro, migrating NC cells in dark and white embryos prefer HA to fibronectin. In vivo, such an HA migration stimulating effect might exist as well, but be counteracted to differing degrees in dark and white embryos. The ultrastructural localization of HA was studied by means of transmission electron microscopic immunohistochemistry using HABP and different protocols of standard chemical fixation, cryofixation, embedding, and immunolabeling. The binding reaction of HA to HABP was strong and showed an equal distribution throughout ECM spaces after both standard chemical fixation/freeze substitution and cryofixation. A preference for the somite or subepidermal side was not observed. Following standard fixation/freeze substitution HABP-labeled "honeycomb"-like networks reminiscent of fixation artifacts were more prominent than labeled fibrillar or irregular net-like structures. The latter predominated in adequately frozen specimens following high-pressure freezing/freeze substitution. For this reason fibrillar or irregular net-like structures very likely represent hyaluronan in the complex subepidermal matrix of the axolotl embryo in its native arrangement.

Ambystoma↗

Electron microscopic investigation of water occlusions in intercellular spaces in the inner cortex of lucerne nodules.

It is unclear to what extent oxygen diffusion pathways through the cortex of the nitrogen-fixing zone of indeterminate nodules are liquid filled and whether a blockage of these pathways is involved in varying nodule oxygen permeability to control nitrogenase activity. We examined the proportion of water-filled intercellular spaces of lucerne (Medicago sativa L.) nodules with cryo-scanning electron microscopy. This technique allows for direct observation of water accumulation. Thirty percent of all intercellular spaces in the inner cortex of lucerne nodules were liquid filled. Decreasing the nodule oxygen permeability by detopping of the plant or by increasing the rhizospheric oxygen partial pressure to 80 kPa had no statistically significant effect on the water distribution in the intercellular spaces. Therefore, the hypothesis of a continuous aqueous diffusion barrier in the inner cortex could not be supported. The abundance of glycoproteins in intercellular spaces of the inner cortex was investigated with immunoelectron microscopy. No alteration due to detopping or after increase of the rhizospheric oxygen partial pressure was observed. Therefore, our results do not support the hypothesis of a short-term regulation of oxygen permeability by blockage of diffusion pathways through morphological changes in the cortex region of the nitrogen-fixing zone of lucerne nodules.

Cryoelectron Microscopy↗

Biological ultrastructure as revealed by high resolution cryo-SEM of block faces after cryo-sectioning.

Ultrastructural information was obtained by imaging the block face of high-pressure-frozen cryo-sectioned biological samples in a high-resolution cryo-SEM. Cryo-sectioning leads to a well-defined flat artificial surface in contrast to cryo-fracturing. Typical artefacts of cryo-sections such as compression and crevasses were not visible on the block face. The ultrastructural features known from resin sections and from freeze-fractures could also be found on the block faces. The cytoplasms show particles of different size which most likely represent proteins. The effects of radiation damage could be reduced considerably by applying the double layer coating technique and backscattered electron imaging. High quality cryo-sections are only obtained from vitrified material. Reasonably flat block faces were, however, also obtained from adequately frozen microcrystalline samples, thereby facilitating ultrastructural studies in the frozen hydrated state.

Animals↗

Double-layer coating for field-emission cryo-scanning electron microscopy--present state and applications.

Imaging of fast-frozen samples is the most direct approach for electron microscopy of organic material. It prevents chemical fixation and drying artifacts. Frozen samples can be replicated and imaged in the transmission electron microscope (TEM), or they can be directly visualized in the cryo-scanning electron microscope (cryo-SEM). Double-layer coating combines these two techniques and many of their advantages. With this method, the frozen bulk sample is coated similar to the TEM-replica technique with, for example, a shadow of platinum (at an angle of 45 degrees) and an additional layer of carbon. Then, the sample is cryo-transferred to an SEM equipped with a cold stage and imaged with the material-dependent backscattered electron signal that shows the platinum distribution. With this method, charging artifacts and the effects of beam damage are significantly reduced. Although currently the resolution of the replica technique cannot be surpassed, the method greatly facilitates the processing of brittle, rapidly frozen samples because no replica cleaning is necessary. This makes the method especially suitable for high-pressure frozen samples.

Freeze Fracturing↗

IGF-I and bFGF differentially influence atrial natriuretic factor and alpha-smooth muscle actin expression in cultured atrial compared to ventricular adult rat cardiomyocytes.

In the present study, we compare expression, storage and secretion of the atrial natriuretic factor (ANF) in atrial and ventricular adult rat cardiomyocytes (aARC and vARC) in long-term culture. The influence of insulin-like growth factor-I (IGF-I) and of basic fibroblast growth factor (bFGF) on ANF production and secretion, as well as on the expression of a structural component, alpha-smooth muscle actin (alpha-sm actin), was studied in the two cell types. Antibodies against alpha-ANF were used for immunocytochemical localization of ANF. aARC contained more ANF-granules than vARC, and they were distributed throughout the cell bodies. Quantitative determination of ANF storage and secretion was done by radioimmunoassay (RIA; 125I), and it was demonstrated that aARC stored and secreted ANF 18- and 16-times more, respectively, when compared to vARC. Immuno-electron microscopy confirmed that ANF storing secretory granules were present in both types of cardiomyocytes. Expression of ANF and alpha-sm actin in aARC and vARC responded differently to treatment with either IGF-I or bFGF. In aARC, neither IGF-I nor bFGF had an influence on expression of ANF. In vARC, expression of ANF was downregulated by IGF-I and upregulated by bFGF with regard to both immunoreactivity and message. In contrast to vARC, expression of alpha-sm actin was not affected by IGF-I in aARC, whereas bFGF produced a strong upregulation similar to that found in vARC. Mitogen-activated protein kinases (MAPK) 42 and 44, though, were equally activated by bFGF and IGF-I in both aARC and vARC.

Actins↗