Targeting focal adhesion assembly by ethoxyfagaronine prevents lymphoblastic cell adhesion to fibronectin.

BACKGROUND: Leukemic cell adhesion to proteins of the bone marrow microenvironment provides signals which control morphology, motility and cell survival. We described herein the ability of ethoxyfagaronine (etxfag), a soluble synthetic derivative of fagaronine, to prevent leukemic cell adhesion to fibronectin peptide (FN/V). METHODS: Phosphorylation of fak and pyk2 were evaluated by immunoblotting. Labelled proteins were localized by confocal microscopy. PI 3-kinase activity was evaluated by in vitro kinase assay. RESULTS: Subtoxic concentration of etxfag reduced L1210 cell adhesion to FN/V dependently of ß1 integrin engagement. Etxfag impaired FN-dependent formation of ß1 clustering without modifying ß1 expression at the cell membrane. This was accompanied by a decrease of focal adhesion number, a diminution of fak and pyk2 phosphorylation at Tyr-576, Tyr-861 and Tyr-579, respectively leading to their dissociations from ß1 integrin and inhibition of PI 3-kinase activity. Etxfag also induced a cell retraction accompanied by a redistribution of phosphorylated fak and pyk2 in the perinuclear region and lipid raft relocalization. CONCLUSION: Through its anti-adhesive potential, etxfag, combined with conventional cytotoxic drugs could be potentially designed as a new anti-leukemic drug.

Elastin-derived peptides enhance melanoma growth in vivo by upregulating the activation of Mcol-A (MMP-1) collagenase.

BACKGROUND: Elastin peptides possess several biological activities and in vitro data suggest they could be involved in the early phase of melanoma growth. METHODS: Using diverse in vitro and in vivo techniques (cell proliferation, invasion and migration assays, zymography, western blots, collagen degradation assay, reverse transcription PCR, melanoma allographs and immunohistochemistry), we analysed the effect of elastin-derived peptides (EDPs) on B16F1 melanoma growth and invasion, as well as on the proteolytic systems involved. RESULTS: We found that EDPs dramatically promote in vivo tumour development of B16F1 melanoma, as well as their in vitro migration and invasion. The inhibition of serine proteases and matrix metalloproteinases (MMPs) activities, by aprotinin and galardin, respectively, demonstrated that these enzymes were involved in these processes. However, we found that EDPs did not increase urokinase-type plasminogen activator, tissue-type plasminogen activator or MMP-2 expression and/or activation, neither in vitro nor in vivo. Nevertheless, we observed a strong increase of pro-MMP-9 secretion in EDPs-treated tumours and, more importantly, an increase in the expression and activation of the murine counterpart of MMP-1, named murine collagenase-A (Mcol-A). Moreover, we show that plasminogen system inhibition decreases collagen degradation by this enzyme. Finally, the use of a specific blocking antibody against Mcol-A abolished EDP-induced B16F1 invasion in vitro, showing that this MMP was directly involved in this process. CONCLUSION: Our data show that in vivo, EDPs are involved in melanoma growth and invasion and reinforced the concept of elastin fragmentation as a predictive factor.

Antibiotic therapy of transaxillary augmentation mammoplasty.

BACKGROUND: Capsular contracture is the most common complication and the main cause of dissatisfaction after augmentation mammoplasty, for both the patient and the plastic surgeon. The formation of fibrous tissue around the prosthesis alters the form or the consistency of the implant, thus modifying the breast shape, its contour and its softness. The initial satisfaction with the achieved aesthetical result is then transformed into great dissatisfaction, due to the presence of a shapeless and undesired mass. PATIENTS AND METHODS: The following study considered data collected between 1998 and 2007. Sixty-seven female patients (aged between 35 and 53 years) who suffered from mammary hypotrophy and had undergone submuscular augmentation mammoplasty were enrolled. All the implanted prostheses were round and texturized, with a volume of 250 cm3 to 450 cm3. The patients underwent pre-, intra- and postoperative antibiotic therapy in order to prevent clinical and subclinical infection of the implants. RESULTS: The follow-up ranged from a period of two to nine years. All patients were examined during the first antibiotic administration and again subsequently, after 1, 3, 6 and 12 months, to evaluate the results in terms of capsular contracture. Of all patients, 90% presented a degree I Baker's classification, the remaining 10% a degree II. Not one of the patients treated showed grade III or IV capsular contracture nor was there any need to remove the prosthesis during the examination period. CONCLUSION: It is clear that a main role in capsular contracture is played by the infectious process, with the activation of specific inflammatory cells. Interfering with the infectious process can prevent fibrotic reaction evolving into capsular contracture. Although the process causing capsular contracture is multifactorial, our study showed a favourable response can be achieved when using antibiotic therapy associated with the transaxillary approach.

Dermatofibrosarcoma protuberans: wide local excision vs. Mohs micrographic surgery.

BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is an uncommon tumor of the skin with high rates of local recurrence. It is debated whether Mohs micrographic surgery (MMS) involves lower recurrence rates than wide local excision (WLE). Recent preliminary reports indicate more consistently favorable cure rates with MMS. We report comparative observational data on 41 patients who underwent MMS and 38 who underwent WLE. Their data were then pooled with those available in the medical literature to obtain more precise estimates of recurrence rates with MMS and WLE. METHODS: The records of 79 patients with DFSP who underwent WLE (n=38) or MMS (n=41) in 1990-2005 were reviewed retrospectively. The primary endpoint was tumor recurrence rate. The PubMed database was searched for DFSP case series treated with WLE or MMS, and the recurrence proportions reported for the two separate procedures were pooled. RESULTS: Five of the 38 WLE patients (follow-up=4.8 years) had recurrences (13.2%, 95% CI 4.4-28.1%) as opposed to none (95% CI 0-8.6%) of the 41 MMS patients (follow-up=5.4 years). Pooling of these data with those from the literature yielded 6/463 recurrences for MMS (1.3%, 95% CI 0.5-2.8%) and 288/1394 recurrences for WLE (20.7%, 95% CI 18.6-22.9%). The relative risk of recurrence for WLE vs. MMS patients was 15.9 (95% CI 7.2-35.5). CONCLUSIONS: Significantly lower recurrence rates were recorded in our patients subjected to MMS compared with those treated with WLE. The pooled data also indicated a clear advantage of MMS. There is inconclusive evidence for any advantage of MMS in non-primary cases, while MMS was most effective in treating head and neck tumors. These data may be useful to guide clinicians in the choice of the more appropriate surgical treatment for DFSP patients.

Cryotherapy in the treatment of keloids.

BACKGROUND: Despite their benign nature, keloids may constitute a severe aesthetic and, in some cases, functional problem with important repercussions on patients' quality of life. There is no consensus on keloid treatment and no wholly satisfactory therapy has yet emerged. OBJECTIVE: To assess the efficacy of cryotherapy in the treatment of keloids. METHODS: 135 patients with 166 keloids received cryosurgical treatment between 1990 and 2004. Freeze times and number of sessions varied. Scar volume was measured before and after treatment. Median follow-up was 48.6 months (range 12.4-72.6 months). RESULTS: Of the 166 lesions treated, 79.5% responded very well with a volume reduction of the initial mass of greater than 80% after a median of 3 treatments (range: 1-9). A good result was obtained in 14.5% of lesions, while results were unsatisfactory in 6% of cases. The main adverse effects reported were atrophic depressed scars and residual hypopigmentation (75% of cases). No recurrences arose during the follow-up period (12-72 months). CONCLUSIONS: To date, cryotherapy appears to be the most effective, safe, economic, and easy-to-perform monotherapy to treat keloid lesions and hypertrophic scars.

Number of mast cells in the Harderian gland of the lizard Podarcis sicula sicula (Raf): the annual cycle and its relation to environmental factors and estradiol administration.

The Harderian gland of the lizard Podarcis sicula sicula (Raf) contains connective tissue type mast cells whose numbers vary during the year showing two peaks, one in spring the other in winter. No sex differences are found throughout the year. Thermal and photoperiodic manipulations indicate that only temperature influences mast cell number (MCN) both in winter and in summer but not in spring. In animals exposed to high temperatures in February (but not in May) MCN declined, while exposure to low temperature in July had the opposite effect. Estradiol treatment of the February and April lizards increased MCN, an effect counteracted by the synthetic antiestrogen tamoxifen; in July lizards, this did not occur. In animals exposed to a high temperature in February, estradiol had no effect, as in animals exposed to low temperatures in July. These data suggest that in spring MCN seems to be more responsive to hormonal stimuli rather than external cues (temperature), while in summer MCN is more sensitive to temperature than to hormonal stimuli (estradiol). Both humoral and external factors are concluded to influence mast cell numbers in the Harderian gland of the lizard P. sicula sicula.