Bioactive molecules typically mediate their biological effects through direct physical association with one or more cellular proteins. The detection of drug-target interactions is therefore essential for the characterization of compound mechanism of action and off-target effects, but generic label-free approaches for detecting binding events in biological mixtures have remained elusive. Here, we report a method termed target identification by chromatographic co-elution (TICC) for routinely monitoring the interaction of drugs with cellular proteins under nearly physiological conditions in vitro based on simple liquid chromatographic separations of cell-free lysates. Correlative proteomic analysis of drug-bound protein fractions by shotgun sequencing is then performed to identify candidate target(s). The method is highly reproducible, does not require immobilization or derivatization of drug or protein, and is applicable to diverse natural products and synthetic compounds. The capability of TICC to detect known drug-protein target physical interactions (K d range: micromolar to nanomolar) is demonstrated both qualitatively and quantitatively. We subsequently used TICC to uncover the sterol biosynthetic enzyme Erg6p as a novel putative anti-fungal target. Furthermore, TICC identified Asc1 and Dak1, a core 40 S ribosomal protein that represses gene expression, and dihydroxyacetone kinase involved in stress adaptation, respectively, as novel yeast targets of a dopamine receptor agonist.

IMPORTANCE Smoking, a common lifestyle trait, is considered by many surgeons to be a major risk factor for postoperative complications. However, in the literature on local reconstruction, the association between smoking and the rate of postoperative complications after cutaneous tissue transfer is not well characterized. OBJECTIVETo study the outcomes of flaps and grafts used in Mohs micrographic surgery reconstruction with respect to smoking status and patient-specific and surgery-specific variables.DESIGN, SETTING, AND PARTICIPANTS This retrospective case-control study was conducted at a single tertiary referral center among 1008 patients who underwent Mohs reconstruction repaired by flap or graft between July 1, 2012, and June 30, 2016, and were selected via consecutive sampling. Cases with incomplete records or those in which a single flap or graft was used to repair multiple defects were excluded. Data analysis was performed from September 2017 to January 2018. MAIN OUTCOMES AND MEASURESPostoperative acute and long-term complications. Acute complications included postsurgical infection, dehiscence, hematoma, uncontrolled bleeding, and tissue necrosis that required medical counseling or intervention. Long-term complications included functional or cosmetic outcomes that prompted the patient to request or the surgeon to offer additional intervention. RESULTSOf the 1008 patients included in the study (396 women and 612 men), the median (SD) age was 70 (12) years (range, 21-90 years). A total of 128 patients (12.7%) were current smokers, 385 (38.2%) were former smokers, and 495 (49.1%) were never smokers. On multivariate logistic regression, current smoking (odds ratio [OR], 9.58; 95% CI, 3.63-25.3), former smoking (OR, 3.64; 95% CI,, larger defect size (OR, 2.25; 95% CI,, and the use of free cartilage graft (OR, 8.19; 95% CI, 2.02-33.1) were associated with increased risks of acute complications. For long-term complications, central face location (OR, 25.4; 95% CI,), use of interpolation flap or flap-graft combination (OR, 3.49; 95% CI, 1.81-6.74), larger flap size (OR, 1.42; 95% CI, 1.09-1.87), and basal cell carcinomas or other basaloid tumors (OR, 3.43; 95% CI, were associated with an increased risk, whereas increased age (OR, 0.66 per 10-year interval; 95% CI, 0.54-0.80) was associated with decreased risk.CONCLUSIONS AND RELEVANCE This study suggests that both current and former smokers are at increased risk for acute postsurgical complications but that smoking status is not associated with long-term complications. These findings may allow the surgeon to better quantify the magnitude of risk and provide helpful information for patient counseling.LEVEL OF EVIDENCE 3.

BACKGROUND The trilobed flap is a useful repair option for distal nasal defects. In certain cases, however, the placement of its quaternary defect may risk compression of the internal nasal valve or induction of ectropion. In this study, we propose a modified design of the trilobed flap, which uses unequal external interlobe angles. OBJECTIVE To present the design principles and results of our modified trilobed flap for the reconstruction of difficult distal nasal Mohs defects. MATERIALS AND METHODS Mohs defects of 26 patients were reconstructed (21 with long-term follow-up) using our modified trilobed flap over 1 year. Two independent masked raters graded postoperative photographs for alar symmetry and overall cosmesis. RESULTS Median alar symmetry scores were excellent and overall cosmesis grades were between excellent and very good. CONCLUSION The modified trilobed flap with unequal external angles offers an excellent option for reconstruction of Mohs defect of the distal nose that may not be well-suited for other repairs. Over 1 year, 21 modified trilobed repairs were performed with overall excellent outcomes.

Chondrosarcoma is a malignant cartilage tumor in which there is constitutive activation of Hedgehog-mediated signaling. Pharmacologic agents that inhibit Hedgehog (Hh) signaling have the potential to be used as novel targeted anti-tumor therapies. IPI-926, a novel, selective, molecule that antagonizes the Hh pathway by binding to Smoothened, is currently in clinical trials. The activity of IPI-926 was assessed in human primary chondrosarcoma tumors obtained at surgery from six different donors and grown as subcutaneous xenografts in NOD/SCID mice. Studies were conducted in primary chondrosarcoma xenografts to assess the activity of IPI-926 either at time of implant or in established tumors and to compare the anti-tumor activity of IPI-926 to chemotherapy and targeted agents. Tumor tissue was collected post-treatment at the end of each study for histopathological analysis or for evaluation of expression of Hh pathway genes by RT-PCR for GLI1, PTCH-1, and SMO mRNA. The chondrosarcoma tumor sizes were significantly smaller in the IPI-926 treated groups, compared to either control or chemotherapy-treated groups. There was also less cellularity, with cells appearing more differentiated, in the IPI-926 treated group. The tumors from mice treated with IPI-926 had significantly reduced expression of Hedgehog target genes GLI1 and PTCH1 compared to those from mice treated with vehicle or other chemotherapies. In addition, inhibition of GLI1 and PTCH1 gene expression was detected in the human tumor cells, a finding that has not been previously demonstrated in carcinomas, where the Hedgehog blockade seems to affect primarily the murine-derived stromal cells. As expected, inhibition of Hh target gene expression was also detected in the tumor stroma in these xenografts. Initial results from gene expression profiling of the tumor cells suggest that several additional genes may be affected by IPI-926 treatment. In summary, IPI-926 administration to mice bearing tumors derived from primary human chondrosarcoma tumors results in down-modulation of the Hh pathway in the tumor cells, as demonstrated by evaluation of the Hh-dependent genes GLI1 and PTCH1. Hh pathway gene expression is also inhibited in the tumor stroma. Down-modulation of the Hh pathway with IPI-926 results in inhibition of growth of both newly implanted and established chondrosarcoma tumors. Decreased tumor growth is accompanied by histopathological changes, including loss of cellularity and calcification. Thus, IPI-926 directly targets the Hh pathway in chondrosarcoma tumor cells and results in growth inhibition and changes in the tumor histopathology. These studies provide strong scientific rationale for further evaluation of Hh pathway inhibition with IPI-926 in humans with chondrosarcoma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-380. doi:10.1158/1538-7445.AM2011-LB-380

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