Merging and scoring molecular interactions utilising existing community standards: tools, use-cases and a case study.

The evidence that two molecules interact in a living cell is often inferred from multiple different experiments. Experimental data is captured in multiple repositories, but there is no simple way to assess the evidence of an interaction occurring in a cellular environment. Merging and scoring of data are commonly required operations after querying for the details of specific molecular interactions, to remove redundancy and assess the strength of accompanying experimental evidence. We have developed both a merging algorithm and a scoring system for molecular interactions based on the proteomics standard initiative-molecular interaction standards. In this manuscript, we introduce these two algorithms and provide community access to the tool suite, describe examples of how these tools are useful to selectively present molecular interaction data and demonstrate a case where the algorithms were successfully used to identify a systematic error in an existing dataset.

Influence of additives to the formulation of n.c.a. [¹¹C]PiB on sterile filter performance.

The influence of different additives (PEG 300, PEG 400, PG) to the product solution of [(11)C]PiB was investigated with regard to tracer retention for a number of commonly used sterile filters for aseptic manufacturing of PET-tracers. The effect of the amount of additive with regard to tracer retention and the resulting viscosity of the filtration solution was determined. Recommendations for the individual combinations of filters and amounts of additives suitable for the different filtration methods that are implemented in commercially available synthesis modules are given as well.

Comparison of in vitro and in vivo approaches to studying brain colonization by breast cancer cells.

Brain metastases occur in 20 to 40% of patients with metastatic breast cancer. The process is complex and depends on successful cancer cell evasion from the primary tumor, distribution and survival within the blood stream and cerebral microvasculature, penetration of the blood brain barrier and proliferation within the brain microenvironment. The initial steps of brain colonization are difficult to study in vivo. Therefore, in vitro assays have been developed to mimic this process. Most commonly, in vitro studies of brain colonization focus on tumor cell adhesion to brain endothelial cells and transendothelial migration. We previously investigated breast cancer brain colonization from the blood stream in vivo and defined the time and process of brain entry for five different cancer cell lines in a mouse model. We now investigated if in vitro approaches can reliably emulate the initial steps that determine successful brain colonization in vivo. To this end, we optimized an in vitro model of the vascular blood brain barrier and compared the brain invasion properties of the in vivo characterized cell models with their ability to interact with and penetrate the blood brain barrier model in vitro. Our results show that the in vitro findings correlate only poorly with the vivo results. The limitations of the in vitro approaches are discussed in light of the in vivo processes. We conclude that investigation of mechanisms supporting the earliest steps of breast cancer brain metastasis from the blood stream will depend on in vivo analyses.

[Total knee replacement in haemophilic arthropathy. A clinical and radiological evaluation of 30 patients]. / Kniegelenksendoprothetik bei hämophiler Arthropathie. Klinische und radiologische Verlaufskontrolle mit 30 Patienten.

UNLABELLED: Purpose of this retrospective study was to evaluate our own results after total knee replacement in patients with haemophilia. Patients, material, method: 30 patients with haemophilia who underwent total knee replacement between 1987 and 2005 were included. We used the clinical and radiological Knee Society Score. Furthermore, the Petterson and the Arnold and Hilgartner score were applied. RESULTS: The mean age at the time of surgery was 43.2 (27-66). At the time of follow-up examination the mean age was 51.6 (30-82) years. The mean follow-up was 7.1 (2-20) years. Preoperative, he mean Arnold and Hilgartner score was 4.17 (±0.59) and the mean Petterson-Score was 9±2.29. Compared to the preoperative deficiency in knee function (KSS-Score 88.17±33.58) an improvement with 166.67 (±22.73) points was seen. 1 patient showed an aseptic loosening after 11 years. DISCUSSION: Total knee replacement in patients with haemophilia improves knee function and quality of life. The results of our study represent results in earlier published studies. Compared to a non-haemophilic normal population the rate of perioperative complications was not increased.

Functional results after giant cell tumor operation near knee joint and the cement radiolucent zone as indicator of recurrence.

BACKGROUND: Giant cell tumor of bone near the knee joints is a dilemma for the operating surgeon. Curettage and bone grafting have a high recurrence, whereas wide resection has a reduced recurrence rate with the compromise of limb function. MATERIALS AND METHODS: Thirty-eight patients with histologically proven giant cell tumor near the knee joint were treated. All patients were reviewed with regard to the operative method, recurrence rate, postoperative arthritis and functional results of the joint. In cases of cement filling, the radiolucent zone and the sclerotic rim were assessed as possible markers for recurrence. RESULTS: 14 male and 24 female patients were included in this study (mean age 28 years, range 13-56 years). All patients underwent surgery, 21 patients were treated with a bone cement filling and additional osteosynthesis after curettage. Seventeen patients were filled with cancellous bone or curettage alone. In the group with bone cement filling after curettage, the recurrence rate was 23.8%, whereas a recurrence rate of 52.9% was detected in the group with cancellous bone filling or curettage alone. The average time to recurrence was two years (5 months to 6 years). An increase of the radiolucent zone was seen in 80% of all patients with a recurrence. CONCLUSION: Cement filling after extensive curettage does not increase the recurrence rate and does not induce osteoarthritis, as long as the continuity of articular cartilage is maintained. Patients with giant cell tumor of bone near the knee joint can be treated satisfactorily with intralesional resection and bone cement packing. The extension of the radiolucent zone after bone cement filling is a reliable indicator for a possible local recurrence.

Effect of lumbar disc replacement on the height of the disc space and the geometry of the facet joints: a cadaver study.

In a study on ten fresh human cadavers we examined the change in the height of the intervertebral disc space, the angle of lordosis and the geometry of the facet joints after insertion of intervertebral total disc replacements. SB III Charité prostheses were inserted at L3-4, L4-5, and L5-S1. The changes studied were measured using computer navigation software applied to CT scans before and after instrumentation. After disc replacement the mean lumbar disc height was doubled (p < 0.001). The mean angle of lordosis and the facet joint space increased by a statistically significant extent (p < 0.005 and p = 0.006, respectively). By contrast, the mean facet joint overlap was significantly reduced (p < 0.001). Our study indicates that the increase in the intervertebral disc height after disc replacement changes the geometry at the facet joints. This may have clinical relevance.

Comparison of periprosthetic bone remodelling after implantation of anatomic and straight stem prostheses in total hip arthroplasty.

INTRODUCTION: Total hip arthroplasty changes bone loading conditions in the proximal femur and induces adaptive remodelling of the periprosthetic bone. These remodelling processes depend on many implant-specific qualities, e.g. material and elasticity of the stem. The objective of this study was to investigate the effect of the stem design on periprosthetic bone remodelling after insertion of an anatomic stem with proximal fixation and the direct comparison to a straight stem prosthesis. MATERIALS AND METHODS: In a prospective study, the changes in periprosthetic bone mineral density (BMD) after implantation of 68 CTX-S anatomic and 22 PPF straight stem prostheses were assessed in the first post-operative year by means of DEXA and zone analysis by Gruen (Clin Orthop 141:17-27, 1979) "Modes of failure" of cemented stem-type femoral components: a radiographic analysis of loosening. Furthermore all patients with CTX-S prostheses were monitored in the second post-operative year. The correlation of adaptive bone remodelling and the systemic bone density was also investigated. RESULTS: In the distal one-third of the straight stem prosthesis, a clearly greater, although not significant, hypertrophy of the periprosthetic bone was observed. No differences in the extent of bone loss between the two prostheses in the regions of interest (ROI) of the proximal bone were observed. The greatest decrease in BMD was registered in the medial femoral neck in both groups. Bone atrophy decreased progressively as the ROI moved distally, ending in a slight increase in BMD in the distal ROI. No significant changes in periprosthetic BMD occurred in the second post-operative year. A strong positive correlation in the regions with the greatest BMD decrease with the systemic BMD was ascertained. CONCLUSION: After implanting a CTX-S prosthesis, as opposed to PPF prostheses, a different pattern of periprosthetic bone remodelling with a slighter hypertrophy of the distal periprosthetic parts was observed. This implies that the extensive proximal, more physiological bone loading of the anatomic stem as well as the removal of less bone while implanting the stem reduces the negative effects of unphysiological strain distribution and stress shielding. The BMD loss in the medial proximal neck cannot be avoided with this stem design either. The lack of significant BMD changes in the second post-operative year suggests that a stabilisation of bone remodelling processes occurs.

[Bisphosphonates in osteoporosis therapy. Standards and perspectives]. / Bisphosphonate in der Osteoporosetherapie. Standards und Perspektiven.

Bisphosphonates are potent inhibitors of bone resorption and approved agents for both the prevention and treatment of osteoporosis. Many clinical studies have shown a consistent reduction in the risk of vertebral fractures with the use of these drugs, while others have shown a clear reduction in the risk of hip and other non-vertebral fractures. The bisphosphonates are divided into three generations based on their potency and chemical structure. In general, as a substance group they are well tolerated and, when applied correctly, the toxicity is low. Alternative dosages, e.g. monthly, three times a month or yearly as an oral drug or i.v. would probably improve patient compliance. Clinical studies have also shown the safety and effectiveness of bisphosphonates over a period of 3-7 years.

[The use of bisphosphonates in arthroplasty]. / Anwendungsmöglichkeiten der Bisphosphonate in der Endoprothetik.

The use of bisphosphonates in joint arthroplasty is the latest field of application for these agents. The mechanism of action of bisphosphonates suggests that they may optimize long-term survival of the implant. Most important is their potency in suppressing periprosthetic osteolysis due to the inflammatory foreign body reaction of wear debris, to decrease periprosthetic osteopenia caused by stress-shielding and to improve the osseointegration of cementless metal implants. The present review provides the latest information on definite and presumed mechanisms of action of bisphosphonates and their clinical importance.

Total hip replacement in patients with severe bleeding disorders. A 30 years single center experience.

Arthropathy of the hip is moderate in frequency in haemophiliac patients, but is less common than ankle, knee or elbow arthropathy. We report about our experience with total hip replacement in patients with severe bleeding disorders over a period of 30 years. Between July 1972 and 2002, 15 hips in 13 patients were replaced. The main bleeding disorders were Haemophilia A in ten patients and severe v. Willebrand disease in three patients. The mean follow-up was 132 months (range 12-363). We can demonstrate good long-term results, with only one aseptic loosening after 14 years and one septic loosening after 14 months in an HIV-positive patient. The Harris Hip Score increased from 48 points (32-66) preoperatively to 89 (76-100) postoperatively. In conclusion, total hip replacement performed in a specialised haemophiliac centre is a safe procedure, and results in pain relief and improvement of the quality of life in patients with severe bleeding disorders.

Fracture of ceramic heads in total hip replacement.

After introduction of ceramics in total hip replacement, there have been several studies on wear and fracture of the femoral head component. Though reports on fractures are few, we saw four fractures within 2 months. In all patients, a cementless hip prosthesis by four different surgeons was implanted between 3/2001 and 2/2004. In three patients, a ceramic-on-polyethylene pair and in one, a ceramic-on-ceramic pair was used. Only one patient suffered an adequate trauma. The mean survival of the ceramic head was 27 months (11-42). In two patients with polyethylene inlays, the inlay showed signs of wear out due to the fractured head. All four revision surgeries had a good outcome with satisfying results and no complications. Though we observe the postoperative development after implantation of ceramic components closely, we still believe that ceramics in total hip replacement in young and active patients are indicated with good long term results.

Erythroid progenitor renewal versus differentiation: genetic evidence for cell autonomous, essential functions of EpoR, Stat5 and the GR.

The balance between hematopoietic progenitor commitment and self-renewal versus differentiation is controlled by various transcriptional regulators cooperating with cytokine receptors. Disruption of this balance is increasingly recognized as important in the development of leukemia, by causing enhanced renewal and differentiation arrest. We studied regulation of renewal versus differentiation in primary murine erythroid progenitors that require cooperation of erythropoietin receptor (EpoR), the receptor tyrosine kinase c-Kit and a transcriptional regulator (glucocorticoid receptor; GR) for sustained renewal. However, mice defective for GR- (GR(dim/dim)), EpoR- (EpoR(H)) or STAT5ab function (Stat5ab(-/-)) show no severe erythropoiesis defects in vivo. Using primary erythroblast cultures from these mutants, we present genetic evidence that functional GR, EpoR, and Stat5 are essential for erythroblast renewal in vitro. Cells from GR(dim/dim), EpoR(H), and Stat5ab(-/-) mice showed enhanced differentiation instead of renewal, causing accumulation of mature cells and gradual proliferation arrest. Stat5ab was additionally required for Epo-induced terminal differentiation: differentiating Stat5ab(-/-) erythroblasts underwent apoptosis instead of erythrocyte maturation, due to absent induction of the antiapoptotic protein Bcl-X(L). This defect could be fully rescued by exogenous Bcl-X(L). These data suggest that signaling molecules driving leukemic proliferation may also be essential for prolonged self-renewal of normal erythroid progenitors.

Management of haemophilic patients with inhibitors in major orthopaedic surgery by immunadsorption, substitution of factor VIII and recombinant factor VIIa (NovoSeven): a single centre experience.

Inhibitors of factor VIII or FIX in haemophilic patients are a common and serious complication associated with an increased risk of life-threatening bleeding during elective surgery. Substitution therapy fails to be effective, therefore an alternative treatment is needed. We have performed six major elective orthopaedic interventions in four patients with haemophilia A and inhibitors. A preoperative immunadsorbant therapy with Therasorb to eliminate inhibitors was successful in four cases, but during FVIII substitution inhibitors increased on day 4 to day 6 after surgery, leading to decreasing FVIII levels. Therefore, therapy was changed to recombinant FVIIa (rFVIIa; NovoSeven). Two interventions had to be covered with sole rFVIIa therapy as immunadsorbant therapy failed to be effective in one case and the need for acute intervention did not allow pretreatment in the other. We did not see increased bleeding during or after surgery when compared to our experience with non-inhibitor haemophilic patients. In conclusion, a preoperative decrease of inhibitors from immunadsorbant therapy, perioperative substitution of FVIII and changing treatment to rFVIIa when inhibitors are increased, is a safe and economic therapy for guaranteeing haemostasis in major elective orthopaedic surgery. On the contrary, sole therapy with rFVIIa allows immediate surgical intervention without a long hospital stay prior to surgery and a need for laboratory monitoring of inhibitor titres and FVIII levels. Our findings support data previously published.

High frequency of DAZ1/DAZ2 gene deletions in patients with severe oligozoospermia.

Deletions of the DAZ gene family in distal Yq11 are always associated with deletions of the azoospermia factor c (AZFc) region, which we now estimate extends to 4.94 Mb. Because more Y gene families are located in this chromosomal region, and are expressed like the DAZ gene family only in the male germ line, the testicular pathology associated with complete AZFc deletions cannot predict the functional contribution of the DAZ gene family to human spermatogenesis. We therefore established a DAZ gene copy specific deletion analysis based on the DAZ-BAC sequences in GenBank. It includes the deletion analysis of eight DAZ-DNA PCR markers [six DAZ-single nucleotide varients (SNVs) and two DAZ-sequence tag sites (STS)] selected from the 5' to the 3'end of each DAZ gene and a deletion analysis of the gene copy specific EcoRV and TaqI restriction fragments identified in the internal repetitive DAZ gene regions (DYS1 locus). With these diagnostic tools, 63 DNA samples from men with idiopathic oligozoospermia and 107 DNA samples from men with proven fertility were analysed for the presence of the complete DAZ gene locus, encompassing the four DAZ gene copies. In five oligozoospermic patients, we found a DAZ-SNV/STS and DYS1/EcoRV and TaqI fragment deletion pattern indicative for deletion of the DAZ1 and DAZ2 gene copies; one of these deletions could be identified as a 'de-novo' deletion because it was absent in the DAZ locus of the patient's father. The same DAZ deletions were not found in any of the 107 fertile control samples. We therefore conclude that the deletion of the DAZ1/DAZ2 gene doublet in five out of our 63 oligozoospermic patients (8%) is responsible for the patients' reduced sperm numbers. It is most likely caused by intrachromosomal recombination events between two long repetitive sequence blocks (AZFc-Rep1) flanking the DAZ gene structures.

[Historical development of sports orthopedics]. / Geschichtliche Entwicklung der Sportorthopädie.

The history of German sports medicine was decisively influenced by the surgeon August Bier at the beginning of the twentieth century. Initially, general medical and physiological problems were emphasized. Individual treatment of injured athletes played an increasing role at the end of last century. Operative treatment of injured athletes and earlier rehabilitation has changed therapeutic standards of orthopedic treatment. During critical discussion of the historical development of the treatment of ruptured anterior cruciate ligaments and meniscal tears, it becomes evident that treatment methods once rejected may now prove to be useful and correct.

Influence of employment characteristics on postpartum mothers' health.

BACKGROUND/PURPOSE: Over half of mothers with infants under one year are employed. This study explored the work experiences of women who returned to employment during the first year and the relationship of employment characteristics to maternal health. DESIGN/METHODS: Longitudinal, repeated measures during pregnancy and 1,4, 8, and 12 months postpartum. Data on employment characteristics and health status gathered between 1990-95 by questionnaire from 149 employed, partnered women residing in a large urban area in the northwestern United States. RESULTS: Work-family interference increased significantly between pregnancy and each postpartum occasion (p <.001). Between 19-34% of the variance in health status at each measurement occasion was explained by employment characteristics. Work-family interference consistently contributed to the variance in health status. CONCLUSIONS: New models are needed to further understand the complex interplay of work and family lives.

Disruption of an inner arm dynein heavy chain gene results in asthenozoospermia and reduced ciliary beat frequency.

Impaired ciliary and flagellar functions resulting in male infertility and recurrent respiratory tract infections are found in patients suffering from primary ciliary dyskinesia (PCD). In most cases, axonemal defects are present, i.e. PCD patients often lack inner and/or outer dynein arms in their sperm tails and cilia, supporting the hypothesis that mutations in dynein genes may cause PCD. However, to date it is unclear whether mutations in dynein heavy chain genes are responsible for impaired flagellar and ciliary motility in mammals. To elucidate the role of the mouse dynein heavy chain 7 (MDHC7) gene, which encodes a component of the inner dynein arm, we have generated mice lacking this dynein heavy chain isoform. Both MDHC7(+/-) and MDHC7(-/-) mice are viable and show no malformations; however, homozygous males produce no offspring. In comparison to MDHC7(+/-) and wild-type mice the spermatozoa of MDHC7(-/-) mice revealed a dramatic reduced straight line velocity and progressive movement, resulting in the inability of MDHC7-deficient sperm to move from the uterus into the oviduct. Additionally, we measured the beat frequency of tracheal cilia and observed a decrease in the beat frequency of approximately 50% in MDHC7(-/-) mice. The reduction in both ciliary and flagellar motility is not correlated with any gross defects in the axonemal structure. The phenotype of MDHC7(-/-) mice is similar to that observed in some patients suffering from PCD, and our data strongly suggest that in some patients this disease could be due to mutations in the homologous human gene DNAH1 (HDHC7).

Stu2 promotes mitotic spindle elongation in anaphase.

During anaphase, mitotic spindles elongate up to five times their metaphase length. This process, known as anaphase B, is essential for correct segregation of chromosomes. Here, we examine the control of spindle length during anaphase in the budding yeast Saccharomyces cerevisiae. We show that microtubule stabilization during anaphase requires the microtubule-associated protein Stu2. We further show that the activity of Stu2 is opposed by the activity of the kinesin-related protein Kip3. Reexamination of the kinesin homology tree suggests that KIP3 is the S. cerevisiae orthologue of the microtubule-destabilizing subfamily of kinesins (Kin I). We conclude that a balance of activity between evolutionally conserved microtubule-stabilizing and microtubule-destabilizing factors is essential for correct spindle elongation during anaphase B.

Isolation and identification of sperm membrane antigens recognized by antisperm antibodies, and their possible role in immunological infertility disease.

Antisperm antibodies (ASA) are the main cause of immunological infertility, as they impair sperm function by binding to the sperm membrane. In this study, we isolated highly enriched sperm membrane proteins by two-dimensional (2D) gel electrophoresis. Isoelectric focusing, as a first dimension, was performed on precast DryStrip IPG 4-7. The second dimension was carried out on 12% sodium dodecyl sulphate-polyacrylamide gels. A total of 18 antigens were identified by the subsequent 2D Western blotting using ASA from seminal plasma samples of infertile patients. Six of the recognized proteins were isolated and analysed by means of mass spectrometry and peptide matching. They were identified as heat shock proteins HSP70 and HSP70-2, the disulphide isomerase ER60, the inactive form of caspase-3 and two subunits of the proteasome (component 2 and zeta chain). The biochemical identification of these proteins will be helpful in understanding the mechanisms by which ASA impair both sperm function and fertilization. Thus, these proteins may also be useful in the development of reliable methods for ASA detection.

Integrin activation controls metastasis in human breast cancer.

Metastasis is the primary cause of death in human breast cancer. Metastasis to bone, lungs, liver, and brain involves dissemination of breast cancer cells via the bloodstream and requires adhesion within the vasculature. Blood cell adhesion within the vasculature depends on integrins, a family of transmembrane adhesion receptors, and is regulated by integrin activation. Here we show that integrin alpha v beta 3 supports breast cancer cell attachment under blood flow conditions in an activation-dependent manner. Integrin alpha v beta 3 was found in two distinct functional states in human breast cancer cells. The activated, but not the nonactivated, state supported tumor cell arrest during blood flow through interaction with platelets. Importantly, activated alpha v beta 3 was expressed by freshly isolated metastatic human breast cancer cells and variants of the MDA-MB 435 human breast cancer cell line, derived from mammary fat pad tumors or distant metastases in severe combined immunodeficient mice. Expression of constitutively activated mutant alpha v beta 3(D723R), but not alpha v beta 3(WT), in MDA-MB 435 cells strongly promoted metastasis in the mouse model. Thus breast cancer cells can exhibit a platelet-interactive and metastatic phenotype that is controlled by the activation of integrin alpha v beta 3. Consequently, alterations within tumors that lead to the aberrant control of integrin activation are expected to adversely affect the course of human breast cancer.