Direct quantitative detection of host cell residual DNA in recombinant Filgrastim by qPCR.

Host cell residual DNA is considered as an impurity in recombinant biopharmaceuticals. This study aimed to develop a direct qPCR method to quantify E. Coli residual DNA in recombinant Filgrastim. The specific primers were designed to amplify E. Coli's 16S-rDNA genomic region, which encodes the 16S-rRNA. The developed qPCR method showed that the designed primer has specifically amplified the target genome without any secondary reaction. The designed primer was also able to amplify the target gene as a representative of residual DNA in the drug matrix. Results show that the amount of residual DNA in Filgrastim is undetectable.

The Anterolateral Thigh Flap as the Flap of Choice for Scalp Reconstruction.

INTRODUCTION: Large scalp soft tissue defects can present difficulties with reconstruction. The ideal flap for scalp reconstruction has yet to be described although the latissimus dorsi flap is frequently referred to as the first choice in this setting. PATIENTS AND METHODS: Following institutional review board approval, the authors reviewed their experience in scalp reconstruction for the past 4 years. Patient demographics, reconstruction indication, flap choice, complications, and outcomes were recorded. RESULTS: Thirteen patients underwent scalp reconstruction with an anterolateral thigh (ALT) free flap. In most patients, the indication was resection of a cutaneous malignancy. In all but 1 patient the facial or more proximal vessels were used for anastomosis. None of the patients required vein grafts to increase pedicle length. The median flap surface area was 156 cm. One flap had vascular compromise. All donor sites healed without complications. DISCUSSION: The ALT flap can emerge as the flap of choice for scalp reconstruction, even when proximal neck vessels are used as the recipient targets. Using a suprafascial dissection and extending the vascular pedicle to the profunda femoris artery can optimize its role in this setting. The ALT flap provides excellent cosmesis and durable scalp coverage with minimal donor site morbidity.

Predictors of Mastectomy Flap Necrosis in Patients Undergoing Immediate Breast Reconstruction: A Review of 718 Patients.

BACKGROUND: Mastectomy flap necrosis (MFN) after mastectomy and immediate breast reconstruction can compromise postsurgical recovery, lead to additional surgeries, and compromise aesthetic outcome. The objective of this study was to determine if there is a difference in the rate of MFN in patients undergoing immediate alloplastic versus immediate autologous breast reconstruction. The secondary objective was to identify additional patient and surgical factors that may influence the rate of MFN. METHODS: A retrospective chart review of patients who underwent immediate breast reconstruction between 2003 and 2011 in the University of British Columbia Breast Program was performed. Demographic, oncologic, reconstructive, and surgical data were compiled. RESULTS: Approximately 404 alloplastic and 314 autologous patients were reviewed. The overall rate of MFN was 12.9%. There was a trend toward a higher MFN rate in the autologous patient group (15.2% vs 11.6%, P = 0.095). After controlling for age, body mass index (BMI), smoking status, preoperative breast radiation, surgery duration, cancer side, mastectomy type, and postoperative chemotherapy, no association was found between reconstruction type and MFN. BMI greater than 30, smoking status, and preoperative radiation were independent predictors of MFN. Surgical factors including longer duration of surgery and Wise pattern mastectomy incision were also found to be associated with increased odds of MFN. CONCLUSION: We found no difference in the rate of MFN when comparing immediate alloplastic and autologous reconstruction methods. A number of patient and surgical factors were found to be predictors of MFN. The results of this large, retrospective study will help surgeons to tailor their reconstruction based on a patient's risk factors to minimize the incidence of MFN.

A Novel Validated Breast Aesthetic Scale.

BACKGROUND: Breast aesthetics impacts patients' quality of life after breast reconstruction, but patients and surgeons frequently disagree on the final aesthetic evaluation. The need for a comprehensive, validated tool to evaluate breast aesthetics independently from the patient motivated this study. METHODS: The 13-item Validated Breast Aesthetic Scale was developed after several internal meetings, and worded to be understood by a nonspecialist. Three items are common for both breasts, with the remaining being side-specific. To test the internal consistency of the scale subitems, postoperative photographs after different breast reconstruction techniques were graded by a six-member panel. To test interrater and intrarater correlation across time, four physicians evaluated the results of abdominally based breast reconstructions following nipple-sparing mastectomies. RESULTS: Graded aesthetic outcomes of 53 patients showed that the Cronbach alpha of the subitems of the scale was 0.926, with no single item that, if excluded, would increase it. Twenty-two patients underwent aesthetic outcomes grading at four different time points. The mean overall appearance was 3.71 ± 0.62. The mean grade for overall nipple appearance was 4.0 ± 0.57. The coefficient alpha of the panel overall aesthetic grade across different time points was 0.957; whereas intragrader reliability for graders 1 through 4 individually showed alpha coefficients of 0.894, 0.9, 0.898, and 0.688, respectively. Similar results were found for the other items of the scale. CONCLUSIONS: The proposed aesthetic scale evaluates different aspects of the breast reconstruction aesthetic result with excellent internal consistency among its subitems. Grading by a gender-balanced, diverse four-member panel using postoperative photographs showed higher reliability and reproducibility compared to single graders.

A survey of current practice patterns in prophylaxis against venous thromboembolism (VTE) and gastrointestinal (GI) ulceration among Canadian burn centers.

Prospective data on efficacy of routine thromboprophylaxis in burn population remains limited. We believe that this uncertainty has lead to diverse management practices across Canada. Similarly, despite data supporting effectiveness of early enteral nutrition (EEN) for gastrointestinal (GI) ulcer prophylaxis, we hypothesize that many burn centers continue to use additional medical prophylaxis. A questionnaire was sent to 16 Canadian burn units regarding their practices of venous thromboembolism (VTE) and GI ulcer prophylaxis. We had 50% response rate. Fifty percent of respondents reported routine use of VTE prophylaxis in all their burn patients regardless of risk factors, 75% of these were among the largest burn centers in Canada. Only 1 center reported use of low molecular weight heparin, Enoxaparin, as their only mode of prophylaxis. With regards to GI ulcer prophylaxis, 62.5% of respondents indicated limiting use of ulcer prophylactic medications to ICU patients. Three (37.5%) centers reported practicing EEN for prophylaxis, 1 of which administered it as the sole modality. 7 of 8 centers used additional pharmacologic prophylaxis, most commonly an H2-blocker, ranitidine. There remains lack of consensus among Canadian burn centers in areas of VTE and GI ulcer prophylaxis, reflecting the limited prospective data in these fields.