[Survival factors in the treatment of hereditary retinal degeneration]. / Uberlebensfaktoren in der Therapie erblicher Netzhautdegenerationen.

Hereditary retinal degeneration is characterized by apoptotic photoreceptor loss, a process governed by intricate molecular interplay and initiated when proapoptotic signals predominate in the individual cell. Identification of molecules involved and their actions has paved the way for testing the ones with anti-apoptotic functions in models of inherited retinal degeneration. Many of these factors are able to slow the course of the degeneration. However, to date no such treatment has been able to stop or even prevent the devolution of the disorder. Moreover, preservation of morphology does not necessarily correlate with preservation of ERG function. Deepened understanding of the pro- and anti-apoptotic networks is clearly needed for survival factors to be feasible for therapy in humans. In comparison, in a dog model of Leber's congenital amaurosis gene therapy could establish retinal function, thus supplying proof of efficacy of the method.

[Innovative implants -- a prerequisite for biological osteosynthesis]. / Neue Implantate als Voraussetzung biologischer Osteosynthesen.

Biological osteosynthesis is a new paradigm in fracture fixation. In contrast to the conservative mechanical operative period, where exact anatomical and rigid fracture fixation was the ultimate ambition, the new perception in fracture management attaches more importance to the conservation of bone perfusion than to the exact anatomical reduction. Because soft tissue condition and local blood circulation are most important for bone reconstruction and healing of fractures, additional damage of soft tissue caused by operation should ideally be avoided. Consequently surgeons aim at minimally invasive operating techniques using no-touch bone implants. This shift in point of view has also been accommodated by a rapid progress in development of innovative bone implants. Solid medullar nails, novel designs and applications of plates, locking screws and biocompatible titanium implants are some of the implantation techniques which pushed the new concept of biological fracture fixation.

Locking Compression Plate (LCP). An osteosynthesis plate based on the Dynamic Compression Plate and the Point Contact Fixator (PC-Fix).

Based on many years of experience with compression plating and promising results obtained with so-called internal fixators, an implant system was developed which combines the two treatment methods in one implant. Despite the combination of these different fracture treatment techniques, no compromises were made with regard to application as a compression plate or as a bridging device in the form of an internal fixator. The individual construction elements in this new implant system have individually proven themselves extremely valuable in clinical practice. The integration of these well proven elements into the new implant system has made it compatible with the majority of existing instruments and conventional screws. The surgeon is free to select the best treatment method to suit the fracture situation and to make combinations as and when necessary. The LCP (Locking Compression Plate), the product of these combinations, is in line with the latest plating techniques, the aim of which is to achieve the smallest possible surgical incisions and to preserve the blood supply to the bone and the adjacent soft tissues.

Treating forearm fractures using an internal fixator: a prospective study.

Some major complications of internal fixation with plates, such as infections and disturbance of healing, have been shown to be related to necrosis of bone and to the soft tissues immediately deep to the plate. This is attributable to plate contact. To deal with this phenomenon, an internal fixator, the Point Contact Fixator, was developed according to a new concept. The Point Contact Fixator resembles a plate but functions like a fixator, that is, the fracture is stabilized using a splint fixed to the bone by monocortical, angularly locked screws that are designed not to exert pressure between the splint and the bone, thereby minimizing implant-to-bone contact. Vascular damage to the osseous blood supply consequently is avoided. The new internal fixator is the first of a new family of implants in addition to nails, plates, and external fixators. To study the potential of the Point Contact Fixator in a prospective study, 79 forearm fractures in 55 patients were treated in a consecutive series by one surgeon using the same technique throughout. Followup to union is reported for 100% of the patients. Handling the fixator was simple; healing was uneventful; and the rate of complication was low.

Prions and the lymphoreticular system.

Following intracerebral or peripheral inoculation of mice with scrapie prions, infectivity accumulates first in the spleen and only later in the brain. In the spleen of scrapie-infected mice, prions were found in association with T and B lymphocytes and to a somewhat lesser degree with the stroma, which contains the follicular dendritic cells (FDCs) but not with non-B, non-T cells; strikingly, no infectivity was found in lymphocytes from blood of the same mice. Transgenic PrP knockout mice expressing PrP restricted to either B or T lymphocytes show no prion replication in the lymphoreticular system. Therefore, splenic lymphocytes either acquire prions from another source or replicate them in dependency on other PrP-expressing cells. The essential role of FDCs in prion replication in spleen was shown by treating mice with soluble lymphotoxin-beta receptor, which led to disappearance of mature FDCs from the spleen and concomitantly abolished splenic prion accumulation and retarded neuroinvasion following intraperitoneal scrapie inoculation.

The development of the distal femur Less Invasive Stabilization System (LISS).

Fractures around the knee typically require operative fixation to achieve an acceptable, functional outcome. The idea behind the Less Invasive Stabilization System (LISS) was to combine the advantages of both interlocked intramedullary nailing techniques and the early advances of the so-called biological plating technique into one system. This paper introduces the mechanical concept of a locked internal fixator and details some important aspects of the anatomical and biomechanical development of the LISS.

Studies on prion replication in spleen.

Some of the early events following scrapie infection take place in the lymphoreticular system (LRS) and result in significant replication of prions in lymphoid organs. The identity of the cells in the LRS that produce prions and their role in neuroinvasion are still unknown. We find that in the spleen of scrapie-infected mice, prions are associated with T and B cells and to a somewhat lesser degree with the stroma, which contains the follicular dendritic cells (FDC's); curiously, no infectivity was found in lymphocytes from blood of the same mice. Thus, splenic lymphocytes either replicate prions or acquire them from another source. Studies on PrP knockout mice with ectopic expression of PrP restricted to only B or T lymphocytes suggest that neither of these by themselves are competent for prion replication. To determine whether B and T cells are able to pick up prions from other sources, irradiated wild-type mice were reconstituted with PrP-deficient lymphohaematopoietic stem cells. Following intraperitoneal inoculation of these mice, no infectivity was found on splenic lymphocytes whereas the stroma (comprising the radiation-resistant, PrP-expressing FDC's) contained prions. These results imply that splenic lymphocytes can acquire prions, possibly from FDC's, but only if they express PrP.

Impaired prion replication in spleens of mice lacking functional follicular dendritic cells.

In scrapie-infected mice, prions are found associated with splenic but not circulating B and T lymphocytes and in the stroma, which contains follicular dendritic cells (FDCs). Formation and maintenance of mature FDCs require the presence of B cells expressing membrane-bound lymphotoxin-alpha/beta. Treatment of mice with soluble lymphotoxin-beta receptor results in the disappearance of mature FDCs from the spleen. We show that this treatment abolishes splenic prion accumulation and retards neuroinvasion after intraperitoneal scrapie inoculation. These data provide evidence that FDCs are the principal sites for prion replication in the spleen.

Neuroinvasion of prions: insights from mouse models.

The prion was defined by Stanley B. Prusiner as the infectious agent that causes transmissible spongiform encephalopathies. A pathological protein accumulating in the brain of scrapie-infected hamsters was isolated in 1982 and termed prion protein (PrPSc). Its cognate gene Prnp was identified more than a decade ago by Charles Weissmann, and shown to encode the host protein PrP(C). Since the latter discovery, transgenic mice have contributed many important insights into the field of prion biology, including the understanding of the molecular basis of the species barrier for prions. By disrupting the Prnp gene, it was shown that an organism that lacks PrP(C) is resistant to infection by prions. Introduction of mutant PrP genes into PrP-deficient mice was used to investigate the structure-activity relationship of the PrP gene with regard to scrapie susceptibility. Ectopic expression of PrP in PrP knockout mice proved a useful tool for the identification of host cells competent for prion replication. Finally, the availability of PrP knockout mice and transgenic mice overexpressing PrP allows selective reconstitution experiments aimed at expressing PrP in neurografts or in specific populations of haemato- and lymphopoietic cells. The latter studies have allowed us to clarify some of the mechanisms of prion spread and disease pathogenesis.

Scrapie pathogenesis in subclinically infected B-cell-deficient mice.

Prion infections can present without clinical manifestations. B-cell deficiency may be a model for subclinical transmissible spongiform encephalopathy, since it protects mice from disease upon intraperitoneal administration of scrapie prions; however, a proportion of B-cell-deficient mice accumulate protease-resistant prion protein in their brains. Here, we have characterized this subclinical disease. In addition, we have studied the possibility that a neurotoxic factor secreted by B cells may contribute to pathogenesis.

PrP-dependent association of prions with splenic but not circulating lymphocytes of scrapie-infected mice.

An intact immune system, and particularly the presence of mature B lymphocytes, is crucial for mouse scrapie pathogenesis in the brain after peripheral exposure. Prions are accumulated in the lymphoreticular system (LRS), but the identity of the cells containing infectivity and their role in neuroinvasion have not been determined. We show here that although prion infectivity in the spleen is associated with B and T lymphocytes and to a lesser degree with the stroma, no infectivity could be detected in lymphocytes from blood. In wild-type mice, which had been irradiated and reconstituted with PrP-deficient lymphohaematopoietic stem cells and inoculated with scrapie prions, infectivity in the spleen was present in the stroma but not in lymphocytes. Therefore, splenic B and T lymphocytes can either synthesize prions or acquire them from another source, but only when they express PrP.

Intramedullary pressure increase for different commercial and experimental reaming systems: an experimental investigation.

OBJECTIVE: To measure the differences in intramedullary (IM) pressure for commercial reamer systems. DESIGN: IM pressure values for the following systems were measured: AO, Biomet, Howmedica grey reamer, Richards, and Zimmer. To investigate the influence of shaft diameter, the AO reamer head was additionally connected to a small shaft (A6/A7). The pressures were measured in plexiglass tubes filled with a mixture of petroleum jelly and paraffin oil with flow properties at 20 degrees C equivalent to those of bovine medullary fat at 36 degrees C. The reaming assemblies were inserted into the tubes using a materials testing machine at a constant speed. In addition, pressure measurements were made using five pairs of human femora to compare Biomet reamers with the AO reamer with thin, flexible drives (A6/A7). RESULTS: The following pressure distributions were obtained (millimeters of mercury; mean value +/- standard deviation): 9.5-millimeter reamer: low for Biomet (272+/-39); moderate for Richards (810+/-101); and high for Howmedica (990+/-132), AO conventional (1,000+/-97), and Zimmer (1,140+/-183); 13.0-millimeter reamer: low for Biomet (132+/-21), Howmedica (204+/-45), and Zimmer (226+/-33); moderate for AO conventional (474+/-42); and very high for Richards (1,734+/-127). The second worst system (AO conventional: 1,000+/-97) became the second best system by simple reduction of the shaft diameter (A6/A7: 378+/-33). CONCLUSION: A comparison of shaft diameters and pressure increase clearly showed that the system with the thinnest shaft produced the lowest pressure values and vice versa.

PrP expression in B lymphocytes is not required for prion neuroinvasion.

Prion diseases are typically initiated by infection of peripheral sites, as in the case of bovine spongiform encephalopathy, new variant Creutzfeldt-Jakob disease, kuru and most cases of iatrogenic Creutzfeldt-Jakob disease. In mouse scrapie, prion infectivity accumulates in lymphoid organs, and the absence of mature B lymphocytes prevents peripherally administered prions from inducing central nervous system disease. We have now assessed whether expression of the cellular prion protein, PrPc, is required for B lymphocytes to mediate neuroinvasion. We found that repopulation of SCID and Rag-1(-/-) mice with fetal liver cells from either PrP-expressing or PrP-deficient mice and from T-cell deficient mice, but not from B-cell deficient mice, is equally efficient in restoring neuroinvasion after intraperitoneal inoculation of scrapie prions. These results indicate that cells whose maturation depends on B cells or their products, such as follicular dendritic cells, may enhance neuroinvasion. Alternatively, B cells may transport prions to the nervous system by a PrP-independent mechanism.

Transgenic and knockout mice in research on prion diseases.

Since the discovery of the prion protein (PrP) gene more than a decade ago, transgenetic investigations on the PrP gene have shaped the field of prion biology in an unprecedented way. Many questions regarding the role of PrP in susceptibility of an organism exposed to prions have been elucidated. For example mice with a targeted disruption of the PrP gene have allowed the demonstration that an organism that lacks PrPc is resistant to infection by prions. Reconstitution of these mice with mutant PrP genes allowed investigations on the structure-activity relationship of the PrP gene with regard to scrapie susceptibility. Unexpectedly, transgenic mice expressing PrP with specific amino-proximal truncations spontaneously develop a neurologic syndrome presenting with ataxia and cerebellar lesions. A distinct spontaneous neurologic phenotype was observed in mice with internal deletions in PrP. Using ectopic expression of PrP in PrP knockout mice has turned out to be a valuable approach towards the identification of host cells that are capable of replicating prions. Transgenic mice have also contributed to our understanding of the molecular basis of the species barrier for prions. Finally, the availability of PrP knockout mice and transgenic mice overexpressing PrP allows selective reconstitution experiments aimed at expressing PrP in neurografts or in specific populations of hemato- and lymphopoietic cells. Such studies have shed new light onto the mechanisms of prion spread and disease pathogenesis.

[Significance of prion protein in transmission of prions and in pathogenesis of spongiform encephalopathies]. / Die Bedeutung des Prion-Proteins in der Ausbreitung von Prionen und in der Pathogenese der spongiformen Enzephalopathien.

Prion disease or transmissible spongiform encephalopathies are caused by novel pathogens termed prions. Unlike classical infectious agents such as viruses or bacteria, prions lack an independent genome and consist largely if not entirely of an abnormal form of the host-encoded prion protein. How prions multiply is not known. A wealth of experimental evidence supports an essential role for the host-encoded prion protein in susceptibility and pathogenesis of prion diseases and in the propagation and spread of prions. In addition, B lymphocytes have been found to play a crucial role in the neuroinvasiveness of prions.

Pinless external fixation. Indications and preliminary results in tibial shaft fractures.

A major drawback of conventional fixator systems is the penetration of the fixator pins into the medullary canal. The pins create a direct link between the medullary cavity and the outer environment. The new AO pinless fixator bypasses this disadvantage by clamping its trocar points onto the outer cortex without penetrating it. Thus, exposure and consequent contamination of the medullary cavity does not occur. The clinical use of this easily manageable fixator with no drilling requirement is for tibial fractures in which the general and local conditions are poor or the infrastructure of the clinic is inadequate for primary internal stabilization or both. All options for a later conversion to internal fixation remain open. For highly unstable tibial shaft fractures, the pinless fixator can be applied as an additional, minimally invasive, external, locked system to increase the stability of intramedullary nail fixation. The pinless external fixator can be combined favorably with the standard AO tubular system and is a valuable addition to the existing fixator systems.

[Epiperiostal, percutaneous plate osteosynthesis. A new minimally invasive technique with reference to "biological osteosynthesis"]. / Die epiperiostale, perkutane Plattenosteosynthese. Eine minimal-invasive Technik unter dem Aspekt der "biologischen Osteosynthese".

Conservation of bone perfusion, protection of the soft tissue envelope and reduction of systemic stress by strengthening the host defence mechanism are general and essential aspects of a biological osteosynthesis. The minimal invasive operating techniques with the use of technical aids and tricks form the necessary presupposition for successful bone healing with a low complication rate. For an epiperiosteal, percutaneous plate osteosynthesis, the technique using a sliding tip and a manipulation handle is demonstrated.

Use of brain grafts to study the pathogenesis of prion diseases.

For the study of prion neurotoxicity, we used neural-grafting techniques: mice devoid of the normal host prion protein (Prnp% mice) received a neural graft and were intracerebrally infected with mouse prions. The growth and differentiation properties of neural grafts were defined. Growth of embryonic neuroectodermal tissue was optimal at gestational days 12.5-13.5. The blood-brain barrier is reconstituted after 7 weeks in most animals. Scrapie-infected PrPC-expressing grafts develop a severe spongiform encephalopathy and contain proteinase-resistant protein and infectivity. Infected grafts deliver high amounts of prions to the host brain without eliciting disease. Infected grafts show a progressive disruption of the blood-brain barrier. Following intraocular prion inoculation of a transplanted Prnp% mouse, prions do not reach the intracerebral graft, indicating that PrP expression is required for propagation along the optic tract.

A crucial role for B cells in neuroinvasive scrapie.

Although prion proteins are most efficiently propagated through intracerebral inoculation, peripheral administration has caused the diseases kuru, iatrogenic Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy (BSE) and new-variant CJD. The development of neurological disease after peripheral inoculation depends on prion expansion within cells of the lymphoreticular system. Here we investigate the identity of these cells by using a panel of immune-deficient mice inoculated with prions intraperitoneally: we found that defects affecting only T lymphocytes had no apparent effect, but that all mutations that disrupted the differentiation and response of B lymphocytes prevented the development of clinical scrapie. As an absence of B cells and of antibodies correlates with severe defects in follicular dendritic cells, a lack of any of these three components may prevent the development of clinical scrapie. However, we found that scrapie developed after peripheral inoculation in mice expressing immunoglobulins that were exclusively of the M subclass and without detectable specificity for the normal form of the prion PrPC, and in mice which had differentiated B cells but no functional follicular dendritic cells. We conclude that differentiated B cells are crucial for neuroinvasion by scrapie, regardless of the specificity of their receptors.