High turnover of extracellular matrix reflected by specific protein fragments measured in serum is associated with poor outcomes in two metastatic breast cancer cohorts.

Increased extracellular matrix (ECM) formation and matrix metalloprotease (MMP)-mediated ECM degradation are parts of tumorgenesis and generates collagen fragments that are released into circulation. We evaluated the association of specific collagen fragments measured in serum with outcomes in two independent metastatic breast cancer (MBC) cohorts. ELISAs were used to measure C1M (MMP-generated type I collagen fragment), C3M (MMP-generated type III collagen fragment), C4M (MMP-generated type IV collagen fragment), and PRO-C3 (pro-peptide of type III collagen) in pretreatment serum from a phase 3 randomized clinical trial of second-line hormone therapy (HR+, n = 148), and a first-line trastuzumab-treated cohort (HER2+, n = 55). All sites of metastases were included. The collagen fragments were evaluated by Cox-regression analysis for their association with time-to-progression (TTP) and overall survival (OS). In the HR+ cohort, higher C1M and C3M levels (75th percentile cut-off) were associated with shorter TTP; all fragments were associated with shorter OS. In the HER2+ cohort, higher levels of all fragments were associated with shorter TTP; higher PRO-C3 was associated with shorter OS. In multivariate analysis of the HR+ trial for OS, higher levels of all fragments were significant for reduced OS when added separately (C1M HR = 2.1, p < 0.001; C3M HR = 1.8, p = 0.028; C4M HR = 1.8, p = 0.018; PRO-C3 HR = 1.8, p = 0.017); none other clinical covariates were significant. In conclusion, collagen fragments quantified in pretreatment serum was associated with shorter TTP and OS in two independent MBC cohorts receiving systemic therapy. If validated, quantification of ECM remodeling in serum has potential as prognostic and/or predictive biomarkers in MBC.

Synaptojanin 2 is a druggable mediator of metastasis and the gene is overexpressed and amplified in breast cancer.

Amplified HER2, which encodes a member of the epidermal growth factor receptor (EGFR) family, is a target of effective therapies against breast cancer. In search for similarly targetable genomic aberrations, we identified copy number gains in SYNJ2, which encodes the 5'-inositol lipid phosphatase synaptojanin 2, as well as overexpression in a small fraction of human breast tumors. Copy gain and overexpression correlated with shorter patient survival and a low abundance of the tumor suppressor microRNA miR-31. SYNJ2 promoted cell migration and invasion in culture and lung metastasis of breast tumor xenografts in mice. Knocking down SYNJ2 impaired the endocytic recycling of EGFR and the formation of cellular lamellipodia and invadopodia. Screening compound libraries identified SYNJ2-specific inhibitors that prevented cell migration but did not affect the related neural protein SYNJ1, suggesting that SYNJ2 is a potentially druggable target to block cancer cell migration.

Two-year results of open-wedge high tibial osteotomy with fixation by medial plate fixator for medial compartment arthritis with varus malalignment of the knee.

PURPOSE: The purpose of our study was to evaluate the complications, technique-related risks, and the clinical course of patients treated with high tibial osteotomy (HTO) for medial arthritis of the knee with varus malalignment. METHODS: Forty-three of 46 consecutive patients (follow-up, 93.5%) treated with HTO using the TomoFix implant (Synthes, Solothurn, Switzerland) were followed-up for 24 months. Radiographic and clinical data were collected preoperatively as well as 6, 12, and 24 months after surgery using standard instruments (Lysholm and subjective International Knee Documentation Committee score). RESULTS: Excellent and good results were achieved in 67.5% of patients. Thirty-seven patients (86.0%) reported clinical improvement at 24 months compared to preoperative status. Evaluation of the clinical course following HTO revealed a significant increase in function after 12 (P < .01) and 24 (P < .01), but not at 6 months (P = .336) after surgery. A further increase was found between 12 and 24 months (P = .017); 67.5% of the study population returned to their predisease sports activity level at 24 months after surgery. Except for 1 case of intra-articular fracture, no severe intraoperative complications were found. One case of nonunion that demanded additional surgery was observed. CONCLUSIONS: HTO with an open-wedge technique using the TomoFix implant seems to be a safe and efficient procedure. Our data show that postoperative recovery is long, with a majority of patients not reaching a functional end-point by 6 or 12 months. In many patients, further improvement was found after 12 months, which might be related to a removal of the implant. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

A feasibility study of computer-assisted bone graft implantation for tissue-engineered replacement of the human ankle joint.

OBJECTIVE: Computer-assisted graft implantation may contribute to achieving biological joint replacement in the future. The purpose of this experimental study was to evaluate the feasibility and accuracy of a series of computer-assisted graft implantations into human cadaver ankle joints. METHODS: Three-dimensional graft models of virtually planned corresponding tibial and talar defects were created from bovine cancellous bone. A platform for computer-assisted surgery (CAS) was set up to implant the grafts. Registration was performed by pair-point matching with anatomical landmarks. In the case of insufficient registration accuracy, artificial landmarks were used for registration. Eight grafts (four tibial, four talar) were implanted in four human cadaver ankle joints. Postoperative CT was used for outcome analysis. The following criteria of accuracy were defined: macroscopic quality of implant fit; quality of the sagittal and coronar joint surface; and quality of the undersurface of the graft in relation to the base of the defect. RESULTS: No technical complications were observed during computer-assisted graft implantation. Clinically acceptable accuracy was achieved in 6 of 8 graft implantations, with implant failure occurring at the tibial and talar location in one ankle joint. In total, 25 of 32 criteria of accuracy were achieved: 6/8 for macroscopic implant fit; 5/8 for quality of the sagittal joint surface; 7/8 for quality of the coronar joint surface; and 7/8 for quality of the undersurface of the graft. Registration with anatomical landmarks did not achieve sufficient accuracy in 4 of 8 cases, whereas registration with artificial landmarks was successful in all these cases. CONCLUSIONS: We demonstrated the feasibility and accuracy of computer-assisted graft implantation for tissue-engineered replacement of the human ankle joint. However, we cannot recommend the present type of registration by pair-point matching with anatomical landmarks due to the considerable inaccuracies. The focus should be on the improvement of non-invasive registration techniques and methods for evaluating postoperative outcome.

Mortality in patients with pelvic fractures: results from the German pelvic injury register.

BACKGROUND: Pelvic and acetabular fractures are rare injuries and account for approximately 3% to 8% of all fractures. Often the result of high energy blunt trauma, most of the patients sustaining pelvic injuries are at high risk of associated injuries strongly influencing outcome and survival rates. Because of anatomic differences it has been suggested that pediatric pelvic fractures are different injuries as compared with that of adults. However, this has been controversially discussed. Aim of this multicenter register study was to identify similarities and differences between pediatric and adult pelvic trauma and evaluate the influence of changes in medical treatment by comparison of two treatment periods. METHODS: In this multicenter register study, data of 4,291 patients treated from 1991 to 1993 (n = 1,723) or 1998 to 2000 (n = 2,568) for pelvic fractures in one of the 23 participating hospitals were evaluated for age, gender, Injury Severity Score (ISS), Hannover Polytrauma Score (PTS), fracture type (using Tile's classification), peripelvic soft tissue injury, need for emergency measures, mortality, cause of death, and need for operative stabilization. We compared the patients' characteristics of the two treatment periods and pediatric with adult pelvic injuries. Statistical analysis was performed using SAS software. RESULTS: There was no difference in terms of ISS, PTS, and presence of peripelvic soft tissue injuries between the two observation periods. Mortality rate dropped significantly from 7.9% to 5% (p < 0.0001) in the latter treatment period. Death was directly attributed to the pelvic injury in 11% from 1991 to 1993. This rate dropped significantly to 7% in the period from 1998 to 2000 (p = 0.020). A type fractures decreased from 61.1% (1991-1993) to 57.1% (1998-2000) of patients (p = 0.028) and except for these simple fractures there was a significant overall trend toward surgical treatment. Multivariate analysis revealed ISS, PTS, concomitant soft tissue injuries, and need for emergency measures as independent risk factors for death whereas surgical stabilization and treatment in the latter treatment period were associated with an increased survival rate. We found no difference between the adult and the pediatric group in terms of ISS and concomitant peripelvic soft tissue injuries. Children were less likely to receive surgical treatment (19.4% vs. 34.5%, p < 0.0001) but requirement for emergency measures was higher in the pediatric group (17.9% vs. 11.1%, p = 0.033). Moreover, we found no significant differences in mortality between both groups (6.1% vs. 8.2%, p = 0.28). Multivariate analysis showed age

Stabilization of the long head of the biceps tendon in the context of early repair of traumatic subscapularis tendon tears.

BACKGROUND: Tears of the subscapularis tendon commonly are associated with instability of the long head of the biceps tendon. Standard surgical treatment includes tenodesis or tenotomy of the biceps tendon. However, chronic discomfort from spasms and cosmetic disadvantages have been reported following both procedures, while the potential for functional impairments remains controversial. We investigated the outcome of stabilization of the long head of the biceps tendon in the context of early repair of traumatic tears of the subscapularis tendon. METHODS: We performed stabilization of an unstable, structurally intact long head of the biceps tendon in twenty-one patients in the acute phase after a traumatic tear of the subscapularis tendon. The average period from the injury to the surgery was 6.2 weeks. Open tendon stabilization and subscapularis reconstruction were performed with transosseous sutures. The follow-up consisted of clinical examination (with determination of the absolute, age and gender-related, and individual relative Constant scores; clinical evaluation of the long head of the biceps; and subjective determination of shoulder function) and dynamic ultrasound examination. RESULTS: The average follow-up period was 28.4 months. The mean absolute Constant score increased from 26.3 points preoperatively to 79.3 points postoperatively (p < 0.01). The mean age and gender-related Constant score improved from 28.0% to 87.0% (p < 0.01). Seven patients showed clinical symptoms consistent with mild biceps tendinopathy. Using dynamic ultrasound examination, we found two cases of recurrent instability (medial subluxation) of the long head of the biceps tendon. Secondary rupture of the long head of the biceps tendon occurred in one patient, twenty-six months after the surgery. CONCLUSIONS: The functional outcomes of stabilization of the long head of the biceps tendon in the context of early repair of a traumatic tear of the subscapularis tendon were comparable with the results of tenodesis or tenotomy reported in previous studies. The cosmetic results were superior, and chronic discomfort from spasms was not observed. Stabilization of the tendon of the long head of the biceps can be recommended as a treatment option for selected patients and should be discussed as an alternative to tenodesis or tenotomy, particularly in a young patient.

Analysis of trastuzumab and chemotherapy in advanced breast cancer after the failure of at least one earlier combination: an observational study.

BACKGROUND: Combining trastuzumab and chemotherapy is standard in her2/neu overexpressing advanced breast cancer. It is not established however, whether trastuzumab treatment should continue after the failure of one earlier combination. In this trial, we report our experience with continued treatment beyond disease progression. METHODS: Fifty-four patients, median age 46 years, range 25-73 years, were included. We analysed for time to tumour progression (TTP) for first, second and beyond second line treatment, response rates and overall survival. RESULTS: Median time of observation was 24 months, range 7-51. Response rates for first line treatment were 7.4% complete remission (CR), 35.2% partial remissions (PR), 42.6% stable disease > 6 months (SD) and 14.8% of patients experienced disease progression despite treatment (PD). Corresponding numbers for second line were 3.7% CR, 22.2% PR, 42.6% SD and 31.5% PD; numbers for treatment beyond second line (60 therapies, 33 pts 3rd line, 18 pts 4th line, 6 pts 5th line, 2 pts 6th line and 1 patient 7th line) were 1.7% CR, 28.3% PR, 28.3% SD and 41.6% PD respectively. Median TTP was 6 months (m) in the first line setting, and also 6 m for second line and beyond second line. An asymptomatic drop of left ventricular ejection fraction below 50% was observed in one patient. No case of symptomatic congestive heart failure was observed. CONCLUSION: The data presented clearly strengthen evidence that patients do profit from continued trastuzumab treatment. The fact that TTP did not decrease significantly from first line to beyond second line treatment is especially noteworthy. Still, randomized trials are warranted.

EGFR activity in HER-2 over-expressing metastatic breast cancer: evidence for simultaneous phosphorylation of Her-2/neu and EGFR.

Her-2/neu overexpression is an important prognostic parameter in breast cancer patients and has become a response predictor for trastuzumab treatment. Nevertheless, while trastuzumab is highly effective in many Her-2/neu overexpressing tumors, some do not respond. The reason for the differential effect is unknown, but it has been hypothesized that the complex interactions between Her-2/neu and other members of the EGFR family are involved in trastuzumab resistance. We have analyzed the protein expression of Her-2/neu, EGFR, and their activated forms, ptyr-1248 Her-2/neu, ptyr-845 EGFR and ptyr-1173 EGFR, in 57 Her-2/neu overexpressing breast tumors and investigated potential correlations between the receptors. By performing immunohistochemistry on paraffin-embedded tissue sections, we found that ptyr-845 EGFR was significantly co-expressed with Her-2/neu and ptyr-1248 Her-2/neu (p=0.043 and p=0.040, respectively), while ptyr-1173 EGFR was only correlated to Her-2/neu expression (p=0.042). Interestingly, EGFR and its activated forms were all significantly inversely correlated with PgR expression (p=0.011, p=0.033 and p=0.032, respectively), and ptyr-845 EGFR was also inversely correlated with ER expression (p=0.008). While we have previously shown that serum levels of the extracellular component of Her-2/neu are associated with tumoral ptyr-1248 Her-2/neu expression, we did not find a similar relationship between serum EGFR and intratumoral total/activated EGFR. We did, however, observe significantly higher levels of serum EGFR in women with 3+ overexpression of HER-2/neu (p=0.047). Taken together, we have demonstrated the activation pattern of EGFR and Her-2/neu in Her-2/neu overexpressing breast cancer. We suggest that EGFR inhibition might enhance the efficacy of trastuzumab by preventing cross-phosphorylation.

Insulin-like growth factor (IGF)-I and IGF-II serum concentrations in patients with benign and malignant breast lesions: free IGF-II is correlated with breast cancer size.

PURPOSE: Insulin-like growth factors (IGFs) are potent mitogens for breast cancer cells in vitro, and elevated IGF-I serum levels are a risk factor for breast malignancies. This study evaluated IGF-I and IGF-II serum levels in healthy women and in patients with benign and malignant breast lesions and correlated them with tumor size. EXPERIMENTAL DESIGN: Serum levels of the total and unbound fractions of IGF-I and IGF-II were analyzed in 65 patients with benign and malignant breast lesions and in 38 women without breast disease. ELISAs were used to detect serum IGF levels, with (total IGF) or without (free IGF) prior acid-ethanol extraction. RESULTS: Total IGF-I serum concentrations were lower in healthy women than in breast cancer patients (P < 0.001) or patients with benign breast lesions (P = 0.010), but no differences were observed in free IGF-I levels. Conversely, healthy women had higher serum levels of free IGF-II than women with breast lesions (P = 0.003), and the free/total IGF-II ratio was significantly reduced in patients with breast disease (P = 0.001). Although IGF-I or IGF-II serum concentrations of breast cancer patients were similar to those of patients with benign lesions, the size of a malignant tumor was correlated to the ratio free/total IGF-II (P = 0.002). CONCLUSIONS: Malignant breast tumors cannot be distinguished from benign breast lesions by systemic IGF serum levels. However, women with breast lesions have decreased IGF-II concentrations, and free IGF-II levels are clearly correlated to the size of a breast cancer, indicating an involvement in tumor growth.