Sirpa Asko-Seljavaara

Recurrent gains of 1q, 8 and 12 in the Ewing family of tumours by comparative genomic hybridization

Comparative genomic hybridization (CGH) was used to detect copy number changes of DNA sequences in the Ewing family of tumours (ET). We analysed 20 samples from 17 patients. Fifteen tumours (75%) showed copy number changes. Gains of DNA sequences were much more frequent than losses, the majority of the gains affecting whole chromosomes or whole chromosome arms. Recurrent findings included copy number increases for chromosomes 8 (seven out of 20 samples; 35%), 1q (five samples; 25%) and 12 (five samples; 25%). The minimal common regions of these gains were the whole chromosomes 8 and 12, and 1q21-22. High-level amplifications affected 8q13-24, 1q and 1q21-22, each once. Southern blot analysis of the specimen with high-level amplification at 1q21-22 showed an amplification of FLG and SPRR3, both mapped to this region. All cases with a gain of chromosome 12 simultaneously showed a gain of chromosome 8. Comparison of CGH findings with cytogenetic analysis of the same tumours and previous cytogenetic reports of ET showed, in general, concordant results. In conclusion, our findings confirm that secondary changes, which may have prognostic significance in ET, are trisomy 8, trisomy 12 and a gain of DNA sequences in 1q.

Ezrin in primary cutaneous melanoma

Ezrin is a member of the ezrin–radixin–moesin family of proteins that link the actin-containing cytoskeleton to the plasma membrane. Ezrin is also connected to signaling molecules involved in the regulation of cell survival, proliferation and migration. Here, we examined the expression of ezrin in 95 primary cutaneous melanomas and correlated ezrin expression with conventional prognostic factors and biomarkers. From 12 patients metastatic tissue samples were also examined. In addition to ezrin staining, Mib-1 proliferation antigen, p53 and Bcl-2 were evaluated. Ezrin immunoreactivity was seen in most tumors; only 19 (20%) melanomas were negative. A total of 48 (51%) tumors had weak immunoreactivity and 28 (29%) strong immunoreactivity. The intensity of ezrin immunoreactivity was associated with tumor thickness (Breslow, P=0.0008) and with tumor invasion level (Clark, P=0.004), thicker tumors having stronger immunoreactivity. Also, there was a correlation between higher Mib-1 index in tumors and strong ezrin expression. All metastatic samples (n=12) showed positive ezrin immunoreactivity. In univariate analysis of survival, patients (n=76) with positive ezrin immunoreactivity had worse clinical disease behavior than those (n=19) without ezrin immunoreactivity, but the difference was not significant (P=0.19). In multivariate analysis of survival, the ezrin immunoreactivity was not a significant marker. The results indicate that ezrin is expressed in most primary melanomas of the skin and in all metastatic tumors. Ezrin expression correlates with tumor thickness and level of invasion suggesting an association between ezrin expression and tumor progression.

Vascular endothelial growth factor-C gene therapy restores lymphatic flow across incision wounds

Edema and insufficient blood perfusion are common problems in reconstructive surgery. The blood vasculature is reconstructed in microvascular flaps, whereas lymphatic vessel function is lost after surgical incision. Here, we demonstrate that vascular endothelial growth factor C (VEGF‐C) gene transfer can be used to reconstruct a lymphatic vessel network severed by incision of skin flaps. We used adenoviral VEGF‐C gene transfer at the edges of epigastric skin flaps in mice. Our results show that VEGF‐C gene expression results in the formation of anastomoses between the lymphatic vessels of the skin flap and the surrounding lymphatic vasculature. Some spontaneous lymphangiogenesis also took place in the control mice, but the lymphatic vessels generated remained nonfunctional even 2 months postoperatively. In contrast, the VEGF‐C treated mice demonstrated persistent lymphatic vessel function during the 2 month follow‐up despite the transient nature of the adenoviral VEGF‐C gene expression. The restoration of lymphatic function by VEGF‐C in skin flaps provides new tools to promote vascular perfusion and to reduce tissue edema in skin and muscle flaps. These results have important implications for the prevention and treatment of surgically induced secondary lymphedema.

Compressive strength of calcium carbonate and hydroxyapatite implants after bone-marrow-induced osteogenesis

Natural coral and structurally similar porous hydroxyapatite (HA) have been used as bone substitutes. They are not osteoinductive but bone formation can be induced by marrow cells, even in extraosseal sites. In our previous study we induced bone formation in porous coral and HA after having implanted the materials in intramuscular pockets in rat. New bone formed only in HA or coral implants soaked with marrow cells; fibrous tissue ingrowth alone was observed in the controls (without marrow). In the present study we examined the effect of tissue ingrowth on the mechanical properties of coral and HA implants obtained in a similar process to that used before. At 12 weeks the compressive strength of HA was higher in the marrow group than in the controls; it was also higher than that of the wet unimplanted material. The HA blocks did not show resorption. Coral resorbed quickly and lost its compressive strength, which was originally higher than in HA. At three weeks the marrow group was stronger than the control specimens. After six weeks only the marrow group, but not the controls, could be tested. Bone ingrowth seemed to maintain the strength of the coral implant even if it was dissolving. The mechanical strength of both materials was comparable to that of cancellous bone.

Neuroendocrine Carcinoma of the Skin (Merkel Cell Carcinoma): Ultrastructural and Immunohistochemical Demonstration of Neurofilaments

In this study we characterized a skin tumor that grew in the temporal region of a 69-year-old woman. On the basis of tumor morphology, a metastasis from a small cell carcinoma of the lung was initially suggested, but X-ray and bronchoscopic studies were negative. The tumor recurred twice within a year, yet no tumors were found elsewhere in the body. Ultrastructurally, cytoplasmic organelles compatible with neuroendocrine storage granules and perinuclear aggregates of intermediate-sized (8-10 nm) filaments were found in many tumor cells. Indirect immunofluorescence microscopy revealed neurofilament-type intermediate filaments in the tumor cells but no keratin- or vimentin-type filaments. Our results further demonstrate neural properties of this tumor type, which is generally considered to have its origin from Merkel cells, the cutaneous neuroendocrine cells.

Sequelae in the abdominal wall after pedicled or free TRAM flap surgery

Twenty-seven free transverse rectus abdominis musculocutaneous (TRAM) and 16 pedicled TRAM flap breast reconstruction patients were studied for 7 to 41 months (mean, 23 months) postoperatively to compare abdominal sequelae after these two operations. The patient groups were demographically similar; mean age was 47 years in both groups. Subjective grading of the results was similar in both groups. The incidence of minor lower abdominal bulges was higher (44%, 7/16) in the pedicled group than in the free TRAM flap group (4%, 1/27). No hernias were found. Delayed healing of the abdominal scar occurred in 3 free TRAM flap and 1 pedicled TRAM flap patients. Two free TRAM flap (8%) and 7 pedicled TRAM (44%) flap patients had minor edge necrosis of the breast. Trunk strength was tested using an isokinetic device (Lido Multi Joint II), and peak torque for flexion (mean, 111 Nm ± 25 Nm in the free TRAM flap group and 123 Nm ± 28 Nm in the pedicled TRAM flap group) and extension (mean, 144 Nm ± 38 Nm and 167 Nm ± 45 Nm) were measured. No statistical differences occurred between these groups. Sit-up performance was tested and graded from 1 to 6. Both groups performed equally (4.8 and 4.8) and within normal values for this age group. Ultrasonography of the rectus muscles revealed that in the free TRAM flap group, the rectus muscle of the operated side was significantly thinner (cranial segment 6.8 mm vs. 7.8 mm, p < 0.05), thus the harvesting of a segment of muscle below the umbilicus seems to disturb the quality of the entire muscle. The mean size of the muscular defect in the free TRAM flap group was 4.3 x 6.1 cm. In this study no differences in patient satisfaction or trunk strength could be found between free and pedicled TRAM flap patients.

Donor site morbidity of radial forearm flaps : A clinical and ultrasonographic evaluation

Harvesting of a forearm flap based on the radial artery has been thought to cause functional or circulatory problems in the donor hand. Eighteen patients were examined three to 24 months after a radial forearm flap had been raised. The function of both hands was studied for grip strength, mobility of the wrist and elbow joints, and sensitivity of the area served by the superficial radial nerve. The patients were interviewed and the cosmetic result was evaluated. Duplex ultrasonography and colour Doppler ultrasonography of both ulnar arteries were done, and the brachial arteries were measured as controls. Angle-corrected peak flow velocity (cm/s) in the ulnar artery of the donor forearm was significantly increased at the level of the wrist compared with the control forearm (100.9 compared with 73.1 cm/s, p = 0.017), as was the ulnar: brachial peak flow velocity ratio (1.18 compared with 0.76, p = 0.001). The grip strength of the donor hand was weaker by 11.9% (86.5 compared with 72.2 Kp), 10 (56%) had areas of sensory loss over the radial nerve distribution, and seven of the 18 patients complained of cold intolerance. Four patients considered the donor site result so bad that they would not have chosen the operation had they known what the result would look like. The radial forearm flap donor site is not without problems, and the patients must be carefully selected and properly informed preoperatively.

Bone marrow induced osteogenesis in hydroxyapatite and calcium carbonate implants

In this experimental study, blocks of natural coral (calcium carbonate) and its structurally similar derivate in the form of hydroxyapatite (calcium phosphate) were implanted in rat latissimus dorsi muscle with autogenous bone marrow to compare their bone-forming capability. A block without marrow placed in the opposite latissimus muscle served as a control. The animals were killed at 3, 6 and 12 weeks and, in the hydroxyapatite group, also at 24 weeks. The sections were analysed histologically and histomorphometrically. Bone was found only in implants containing bone marrow. Bone formation was significantly (p < 0.05) higher in coral than in hydroxyapatite implants at 3 weeks (10.8% versus 4.8%) and at 12 weeks (13.7% versus 6.3%, bone/total original block area). At 12 weeks all the coral implants had lost their original structure, and the cross-sectional area of the block had diminished to 40% of the original area.

Overrepresentation of 1q21–23 and 12q13–21 in lipoma-like liposarcomas but not in benign lipomas: A comparative genomic hybridization study

Twenty lipomatous tumors, including eight lipoma-like liposarcomas and 12 benign lipomas, were analyzed using comparative genomic hybridization (CGH). DNA sequence copy number changes detected in five lipoma-like liposarcomas (mean, 1.1 aberrations/tumor; range, 0-2) consisted of gains of 12q13-21 (five tumors) and 1q21-23 (four tumors). Two of the tumors showed high-level amplification at 12q14-21 and one tumor at 1q21-22. No copy number changes were found in lipomas. Overrepresentation of 1q and 12q sequences was a recurrent finding in lipoma-like liposarcomas but not in lipomas. Thus, CGH may help in the differential diagnosis of low-grade or borderline adipose neoplasms.

Late results of skin-sparing mastectomy followed by immediate breast reconstruction†

Skin‐sparing mastectomy (SSM) facilitates optimal immediate breast reconstruction (IBR) by preserving the inframammary fold and most of the breast skin. Concerns persist that SSM might increase the rate of local recurrence as the surgical approach is less extensive. Patients who had SSM and IBR over 10 years at a single institution were reviewed.