Full-thickness skin reconstruction with basic fibroblast growth factor-impregnated collagen-gelatin sponge.

INTRODUCTION: This study examined the usefulness of basic fibroblast growth factor impregnated collagen-gelatin sponge (bFGF-CGS) in reconstructive surgery for various acute skin defects including deep dermal burns, facial full-thickness skin defects, and finger amputations as the first clinical application. METHODS: Reconstructive surgery was performed in two stages with bFGF-CGS in 8 male subjects, ranging in age from 6 to 84 years, with acute full-thickness skin defects. Following the adequate debridement of the defect, surgeons prepared a bFGF-CGS with bFGF solution at a dose of 7-14 mg/cm2 approximately 10 min just before application and then secured the bFGF-CGS in place with non-absorbable sutures. Second-stage wound closure was performed with autologous skin grafting following adequate dermis-like tissue regeneration at the site postoperatively. Follow-up was continued for 6 months. RESULTS: Of the 8 subjects, the mean duration from the adequate vascularization of the dermis-like tissue until the second-stage autologous skin graft was 22 ± 4 days. Wound closure was achieved in all cases; the mean duration until wound closure was 32 ± 8 days. During the 6-month follow-up period, no wound infection, recurrent skin ulceration, and no exposure of tendon, bone, and cartilage were observed, and there were no cases of indirectly restricted range of motion from postoperative scar contracture and none with disfiguring scars. CONCLUSION: The authors achieved favorable outcomes following reconstructive surgery with a hybrid artificial dermis impregnated with bFGF for treating acute full-thickness skin defects. bFGF-CGS serves as a convenient regenerative device requiring no specialized medical facilities.

Signature of positive selection in mitochondrial DNA in Cetartiodactyla.

Acceleration of the amino acid substitution rate is a good indicator of positive selection in adaptive evolutionary changes of functional genes. Genomic information about mammals has become readily available in recent years, as many researchers have attempted to clarify the adaptive evolution of mammals by examining evolutionary rate change based on multiple loci. The order Cetartiodactyla (Artiodactyla and Cetacea) is one of the most diverse orders of mammals. Species in this order are found throughout all continents and seas, except Antarctica, and they exhibit wide variation in morphology and habitat. Here, we focused on the metabolism-related genes of mitochondrial DNA (mtDNA) in species of the order Cetartiodactyla using 191 mtDNA sequences available in databases. Based on comparisons of the dN/dS ratio (ω) in 12 protein-coding genes, ATP8 was shown to have a higher ω value (ω = 0.247) throughout Cetartiodactyla than the other 11 genes (ω < 0.05). In a branch-site analysis of ATP8 sequences, a markedly higher ω value of 0.801 was observed in the ancestral lineage of the clade of Cetacea, which is indicative of adaptive evolution. Through efforts to detect positively selected amino acids, codon positions 52 and 54 of ATP8 were shown to have experienced positive selective pressure during the course of evolution; multiple substitutions have occurred at these sites throughout the cetacean lineage. At position 52, glutamic acid was replaced with asparagine, and, at position 54, lysine was replaced with non-charged amino acids. These sites are conserved in most Artiodactyla. These results imply that the ancestor of cetaceans underwent accelerated amino acid changes in ATP8 and replacements at codons 52 and 54, which adjusted metabolism to adapt to the marine environment.

A New Procedure for Wrapped-Negative Pressure Wound Therapy for Congestion After Arterialized Venous Flap Surgery.

Negative pressure wound therapy (NPWT) is a method for treating wound. However, there are no case reports using NPWT for treating congestion after arterialized venous flap. Therefore, this study reported favorable outcomes after using a single-use NPWT system for managing congestion. A 39-year-old man had his index finger caught by a press machine. The finger had a soft tissue defect at the ventral part. An arterialized venous flap taken from the right forearm was transplanted. Perfusion of the flap was favorable, but on postoperative day 5, congestion and the edema of the flap were found. Then, NPWT was initiated. The congestion and edema in the flap were improved without complications such as flap necrosis and wound infection. At 4 months postoperatively, the morphology of the finger was favorable. In this study, NPWT was speculated to force the deeper blood vessels within the flap to dilate with inducing drainage and the simultaneous reduction in excess blood flow to the cortical layer, resulting in the improvement of congestion. Negative pressure wound therapy was used for treating congestion after the transplantation of arterialized venous flap, and the wound was favorably managed.

A Pentagram Suture Technique for Closing Tumor Resection Sites in the Face.

BACKGROUND: Resection of facial skin tumors aims to remove the tumors completely and make the surgical scar unnoticeable as much as possible. By improving the purse string suture method, we developed a new pentagram suture technique that enables simple and safe suturing of small to large defects with early satisfactory esthetic outcomes. The surgical outcomes of a case series were examined in this report. METHODS: As in drawing a unicursal star, 5 suture sites were marked at specific intervals around the defect area. A needle with 5-0 polydioxanone suture was passed from the subcutaneous tissue to the superficial dermal layer at one site and then from the superficial dermal layer to the subcutaneous layer at the next site, and the process was repeated until the pentagram was complete. When apposition was not tight enough, a couple of external stitches were added using 6-0 nylon suture. RESULTS: In 13 patients (16 benign or malignant tumors; mean age, 51.1 years) with a mean tumor size of 10.1 ± 5.2 mm and postoperative skin defect diameter of 12.1 ± 8.2 mm, closure did not result in high tension on the suture, and there was reduced mechanical stress at the wound margin. Surgical outcomes were good esthetically at 6 months after surgery without keloid formation or scar contracture. None of the patients had postoperative pain, infection, or tumor recurrence. CONCLUSIONS: This simple alternative method for the closure of facial skin defects after skin tumor excision could be performed easily and provided satisfactory surgical outcomes.

Degradation of carbohydrate moieties of arabinogalactan-proteins by glycoside hydrolases from Neurospora crassa.

Arabinogalactan-proteins (AGPs) are a family of plant proteoglycans having large carbohydrate moieties attached to core-proteins. The carbohydrate moieties of AGPs commonly have ß-(1→3)(1→6)-galactan as the backbone, to which other auxiliary sugars such as l-Ara and GlcA are attached. For the present study, an α-L-arabinofuranosidase belonging to glycoside hydrolase family (GHF) 54, NcAraf1, and an endo-ß-(1→6)-galactanase of GHF 5, Nc6GAL, were identified in Neurospora crassa. Recombinant NcAraf1 (rNcAraf1) expressed in Pichia pastoris hydrolyzed radish AGPs as well as arabinan and arabinoxylan, showing relatively broad substrate specificity toward polysaccharides containing α-L-arabinofuranosyl residues. Recombinant Nc6GAL (rNc6GAL) expressed in P. pastoris specifically acted on ß-(1→6)-galactosyl residues. Whereas AGP from radish roots was hardly hydrolyzed by rNc6GAL alone, ß-(1→6)-galactan side chains were reduced to one or two galactan residues by a combination of rNcAraf1 and rNc6GAL. These results suggest that the carbohydrate moieties of AGPs are degraded by the concerted action of NcAraf1 and Nc6GAL secreted from N. crassa.

The utilization of a biopsy needle to obtain small muscle tissue specimens to analyze the gene and protein expression.

Recent analytical methods such as real-time polymerase chain reaction (PCR) and Western blotting have now enabled us to analyze the gene and protein expression from small amounts of tissue. A fine needle muscle biopsy is thus expected to obtain a minimally sufficient amount of skeletal muscle to make a successful analysis. As a result, we used this fine needle muscle biopsy technique to obtain muscle tissue specimens from the vastus lateral muscle in 40 participants. The amount of tissue obtained by the fine needle was 5.2 +/- 3.2 mg (mean +/- standard deviation). The total RNA extracted was 3.0 +/- 1.4 microg and the total protein extracted was 2203 +/- 1541 microg. Furthermore, the skeletal muscle tissue specimens obtained by the regular needle technique and blood sample were used as the control. Those specimens were used to measure the gene expression of beta-myosin heavy chain slow (beta-MHC slow) by real-time PCR and the protein expression of monocalboxylate transporter 1 (MCT-1) by Western blotting. Beta-MHC slow gene expression was detected in both samples obtained by a fine and a regular needle biopsy, but not in a blood sample. Furthermore, the MCT-1 protein was detected in samples obtained by a fine needle muscle biopsy. These results indicated that the fine needle muscle biopsy is therefore a useful technique to obtain skeletal muscle specimens at least to analyze the gene and protein expression.

Thyroid hormone targets matrix Gla protein gene associated with vascular smooth muscle calcification.

Thyroid hormones have marked cardiovascular effects in vivo. However, their direct effects on vascular smooth muscle cells have been unclear. Because thyroid hormones play critical roles in bone remodeling, we hypothesized that they are also associated with vascular smooth muscle calcification, one of the pathological features of vascular sclerosis. To test this hypothesis, we examined the effects of 3',3,5-triiodo-L-thyronine (T3) on the expression of calcification-associated genes in rat aortic smooth muscle cells (RAOSMCs). Quantitative RT-PCRs revealed that a physiological concentration of T3 (15 pmol/L free T3) increased mRNA level of matrix Gla protein (MGP), which acts as a potent inhibitor of vascular calcification in vivo, by 3-fold in RAOSMCs, as well as in cultured human coronary artery smooth muscle cells. In RAOSMCs transiently transfected with a luciferase reporter gene driven by the MGP promoter, T3 significantly stimulated luciferase activity. In addition, RNA interference against thyroid hormone receptor-alpha gene diminished the effect of T3 on MGP expression. Aortic smooth muscle tissues from methimazole-induced hypothyroid rats (400 mg/L drinking water; 4 weeks) also showed a 68% decrease in the MGP mRNA level, as well as a 33% increase in calcium content compared with that from the control euthyroid animals, whereas hyperthyroidism (0.2 mg T3/kg IP; 10 days) upregulated MGP mRNA by 4.5-fold and reduced calcium content by 11%. Our findings suggest that a physiological concentration of thyroid hormone directly facilitates MGP gene expression in smooth muscle cells via thyroid hormone nuclear receptors, leading to prevention of vascular calcification in vivo.