Application of Fourier transform infrared spectroscopy to biomolecular profiling of cultured fibroblast cells from Gaucher disease patients: A preliminary investigation.

BACKGROUND: Gaucher disease (GD) is defined as an autosomal recessive disorder resulting from the deficiency of glucocerebrosidase (E.C. 3.2.1.45). Glucocerebrosidase is responsible for the degradation of glucosylceramide into ceramide and glucose. The deficiency of this enzyme results in the accumulation of undegraded glucosylceramide, almost exclusively in macrophages. With Fourier transform infrared (FTIR) spectroscopy, the complete molecular diversity of the samples can be studied comparatively and the amount of the particular materials can be determined. Also, the secondary structure ratios of proteins can be determined by analysing the amide peaks. OBJECTIVES: The primary aim of this study is to introduce FTIR-ATR spectroscopy technique to GD research for the first time in the literature and to assess its potential as a new molecular method. MATERIAL AND METHODS: Primary fibroblast cell cultures obtained from biopsy samples were used, since this material is widely used for the diagnosis of GD. Intact cells were placed onto a FTIR-ATR crystal and dried by purging nitrogen gas. Spectra were recorded in the mid-infrared region between 4500-850 cm-1 wavenumbers. Each peak in the spectra was assigned to as organic biomolecules according to their chemical bond information. A quantitative analysis was performed using peak areas and we also used a hierarchical cluster analysis as a multivariate spectral analysis. RESULTS: We obtained FTIR spectra of fibroblast samples and assigned the biomolecule origins of the peaks. We observed individual heterogeneity in FTIR spectra of GD fibroblast samples, confirming the well-known phenotypic heterogeneity in GD at the molecular level. Significant alterations in protein, lipid and carbohydrate levels related to the enzyme replacement therapy were also observed, which is also supported by cluster analysis. CONCLUSIONS: Our results showed that the application of FTIR spectroscopy to GD research deserves more attention and detailed studies with an increased sample size in order to evaluate its potential in the diagnosis and follow-up of GD patients.

Clinical Experience With the Combination of a Biceps Femoris Muscle Turnover Flap and a Posterior Thigh Fasciocutaneous Hatchet Flap for the Reconstruction of Ischial Pressure Ulcers.

BACKGROUND: The reconstruction of ischial pressure ulcers is problematic because of the distinctive anatomical properties of the region and high recurrence rates. To date, no single technique has been proven to be effective in reducing recurrence of the ulcers. We present our experience with the combination of a biceps femoris muscle turnover flap and a posterior thigh fasciocutaneous hatchet flap and discuss the long-term results. METHODS: A retrospective clinical analysis of 15 patients with grade 4 ischial pressure ulcers reconstructed with biceps femoris muscle turnover flaps and laterally based posterior thigh fasciocutaneous hatchet flaps was carried out between January 2010 and January 2013. Debridement and reconstruction of the ulcers were accomplished in a single stage. The posterior thigh fasciocutaneous flap was elevated in a hatchet style. The long and/or short head of the biceps femoris muscle were dissected from their insertions, turned over on their major pedicles, and their distal portions were used to obliterate the cavitary defect. The skin defect over the muscles was covered by the fasciocutaneous hatchet flap. The average age of the patients was 42.6 years and the mean follow-up time was 27.2 months. RESULTS: Three patients had the following early postoperative complications: hematoma, suture dehiscence, and the necrosis of the short head of biceps muscle. Only 1 patient had a recurrent ulcer 15 months after surgery, which was treated with debridement and the readvancement of the fasciocutaneous flap. The overall recurrence rate was 6.6%. CONCLUSIONS: The biceps femoris muscle turnover flap combined with the posterior thigh fasciocutaneous hatchet flap is a worthwhile option to consider for the reconstruction of ischial pressure ulcers and this technique produces favorable results in terms of the lack of recurrence and complications. The use of the muscle and fasciocutaneous tissue as 2 different flaps, which have different roles in the early and late postoperative period, reduces the recurrence rate.

Reconstruction of traumatic lower extremity soft tissue defects with the combination of ipsilateral and contralateral perforator flaps.

Many flaps have been used to repair complex soft tissue defects of the leg. When the affected areas are relatively small and the local flaps are not suitable, microsurgical operation seems to be an excessive treatment and thus perforator flaps can be used. The authors present the combination of ipsilateral and contralateral perforator flaps for coverage of the traumatic lower extremity.