Judy W. Lee

Inhibition of Smad3 Expression in Radiation-Induced Fibrosis Using a Novel Method for Topical Transcutaneous Gene Therapy

OBJECTIVE To attempt to mitigate the effects of irradiation on murine skin after high-dose radiation using a novel transcutaneous topical delivery system to locally inhibit gene expression with small interfering RNA (siRNA) against Smad3. DESIGN Laboratory investigation. SETTING University laboratory. SUBJECTS Twenty-five wild-type C57 mice. INTERVENTION In an isolated skin irradiation model, the dorsal skin of C57 wild-type mice was irradiated (45 Gy). Just before irradiation, Smad3 and nonsense siRNA were applied to 2 separate dorsal skin areas and then reapplied weekly. Skin was harvested after 1 and 4 weeks. Smad3 expression were assessed by immunohistochemistry, and collagen deposition and architecture was examined using picrosirius red collagen staining. MAIN OUTCOME MEASURES Epidermal thickness was measured semiquantitatively at 4 weeks. Radiation-induced fibrosis was measured quantitatively via tensiometry. The Young modulus, a measure of cutaneous rigidity inversely related to elasticity, was determined, with normal irradiated skin serving as a control specimen. RESULTS Murine skin treated with topical Smad3 siRNA demonstrated effective Smad3 inhibition at 1 week and persistent suppression at 4 weeks. Collagen deposition and epidermal thickness were significantly decreased in skin treated with Smad3 siRNA compared with control irradiated skin. Tensiometry demonstrated decreased tension in Smad3 siRNA-treated skin, with a Young modulus of 9.29 MPa (nonirradiated normal skin, 7.78 MPa) compared with nonsense (control) siRNA-treated skin (14.68 MPa). CONCLUSIONS Smad3 expression can be effectively silenced in vivo using a novel topical delivery system. Moreover, cutaneous Smad3 inhibition mitigates radiation-induced changes in tissue elasticity, restoring a near-normal phenotype.

Techniques for early diagnosis and management of cervicofacial necrotising fasciitis

BACKGROUND Cervicofacial necrotising fasciitis carries high rates of morbidity and mortality, and is not often initially suspected due to its rarity and misleadingly innocuous presentation. We propose an algorithm for the timely diagnosis and management of cervicofacial necrotising fasciitis. METHODS Retrospective review of seven patients ultimately diagnosed with cervicofacial necrotising fasciitis. RESULTS In these seven patients, common presenting symptoms included sore throat, fever and neck pain. On initial examination and imaging, only three had obvious findings. One patients diagnosis was facilitated via a bedside cut-down procedure. Six patients underwent surgical debridement. Four required tracheotomy, and five wounds closed via secondary intention. There were two deaths. CONCLUSION The severity of cervical necrotising fasciitis and its rapid spread necessitate early diagnosis and timely surgical management. The presentation often appears benign. A high index of clinical suspicion should be maintained in cases of neck cellulitis with nonspecific clinical findings, especially in diabetic or otherwise immunocompromised patients. A normal computed tomography scan does not rule out necrotising fasciitis. A cut-down procedure may be critical to early diagnosis in some cases.

Surgical and Nonsurgical Treatments of the Nasal Valves

Nasal obstruction is known to be associated with a major decrease in disease-specific quality of life, and nasal valve dysfunction can play a considerable role in nasal airflow obstruction. Diagnosis and treatment of nasal valve dysfunction requires a thorough understanding of normal anatomy and function as well as pathophysiology of common abnormalities to properly treat the exact source of dysfunction. As the pathophysiology of the nasal valves has become better understood, surgery designed to treat its dysfunction has evolved. Here, we explore the progress we have made in treating the nasal valves, and the deficiencies we still face.

Regulators and mediators of radiation-induced fibrosis: Gene expression profiles and a rationale for Smad3 inhibition

OBJECTIVE: Radiotherapy, an essential modality in cancer treatment, frequently induces fibrotic processes in the skin, including accumulation of extracellular matrix. Transforming growth factor-β is essential in regulating extracellular matrix gene expression and is dependent on Smad3, an intracellular mediator/transcription factor. Our study characterized the genetic expression involved in extracellular matrix accumulation during radiationinduced fibrosis. We performed Smad3 gene silencing in an attempt to abrogate the effects of radiation. STUDY DESIGN: Laboratory research. SETTING: University laboratory. SUBJECTS AND METHODS: C57 murine dermal fibroblasts were irradiated with 20 Gy RNA isolated (0, 6, 12, 24, 48, 72 hours postirradiation) and mRNA analyzed (reverse transcriptase polymerase chain reaction) for known regulators (Smad3, interleu-kin-13 [IL-13]), tumor necrosis factor-α [TNF-α]) and mediators of fibrosis (collagen 1A1 [Col1A1]), TGF-β, matrix metalloprotease-1 and −2 (MMP-1, MMP-2), and tissue inhibitor of metallo-protease-1 (TIMP-1). Smad3 gene expression was silenced using siRNA in an effort to restore an unirradiated gene profile. RESULTS: Following irradiation, there was a steady increase in mRNA expression of Smad3, IL-13, TGF-β, Col1A1, MMP-2, TIMP-1, with peak at 12 to 24 hours and subsequent decline by 72 hours. TNF-α expression remained elevated throughout. MMP-1 showed minimal expression initially, which decreased to negligible by 72 hours. Inhibition of Smad3 significantly decreased expression of Col1A1, TGF-β, MMP-2, and TIMP-1. IL-13 and TNF-α expression was not affected by Smad3 silencing. CONCLUSION: We have characterized the early-phase mRNA expression profiles of the major mediators of radiation-induced fibrosis. Smad3 siRNA effectively abrogated the elevation of Col1A1, TGF-β, TIMP-1, and MMP-2. IL-13 and TNF-α were unaffected by Smad3 silencing and appear to be minor regulators in fibrosis. These findings suggest a therapeutic rationale for Smad3 silencing in vivo.

Trends in functional rhinoplasty 2008

Purpose of reviewThis article reviews the recent literature on functional rhinoplasty for the most important contributions in the field. Recent findingsSurgical techniques for improving the internal nasal valve include upper lateral cartilage fold-in flap, splay graft, alar batten graft, Z-plasty, and the alloplastic Monarch implant. The Nasal Obstruction Septoplasty Effectiveness (NOSE) score and the Rhinoplasty Outcomes Evaluation score have been applied to objectify outcomes in functional rhinoplasty. Functional endoscopic sinus surgery (FESS) and rhinoplasty continue to be safely used in the same surgical sitting. SummaryThe last few years have seen improved perspective on what surgery can do, substantiating the inherent difficulties of establishing reproducible outcomes in form and function of the nose.

Radiation-Induced Fibrosis: A Rationale for Smad3 Inhibition

O R A LS RESULTS: Treatment of HNSCC cells with the inflammatory mediator, IL-1beta, caused the downregulation of E-cadherin expression and upregulation of COX-2 expression. This effect was blocked in the presence of celecoxib. IL-1beta -treated HNSCC cell lines demonstrated a significant decrease in Ecadherin mRNA and an increase in the mRNA expression of the transcriptional repressor Snail. IL-1beta exposure led to enhanced Snail binding at the chromatin level. ShRNA-mediated knockdown of Snail interrupted the capacity of IL-1beta to downregulate E-cadherin. In a SCID xenograft model, HNSCC Snail overexpressing cells demonstrated significantly increased primary and metastatic tumor burdens. CONCLUSIONS: Inflammatory mediators modulate Snail and thereby regulate COX-2-dependent E-cadherin expression in HNSCC. This is the first report indicating the role of Snail in the inflammation-induced promotion of EMT in HNSCC. This newly defined pathway for transcriptional regulation of E-cadherin in HNSCC has important implications for targeted chemoprevention and therapy.