Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

Tokyo Women's Medical University School of Medicine

We developed an inexpensive LIF dual-channel detection system and applied it to a ligase detection reaction (LDR)/CGE method to identify the allelic composition of low-abundance point mutations in a large excess of wild-type DNA in a single reaction with a high degree of certainty. Ligation was performed in a tube with a nonlabeled common primer and multiplex discriminating primers, each labeled with a different standard fluorophore. The discriminating primers were directed against three mutant variations in codon 12 of the K-ras oncogene that have a high diagnostic value for colorectal cancer. LDR products generated from a particular K-ras mutation through successful ligation events were separated from remaining discriminating primers by CGE, followed by LIF detection using the new system, which consists of two photomultiplier tubes, each with a unique optical filter. Each fluorophore label conjugated to the corresponding LDR product produced a distinct fluorescence signal intensity ratio from the two detection channels, allowing spectral discrimination of the three labels. The ability of this system to detect point mutations in a wild-type sequence-dominated population, and to disclose their allelic composition, was thus demonstrated successfully.

Development of a ligase detection reaction/CGE method using a LIF dual-channel detection system for direct identification of allelic composition of mutated DNA in a mixed population of excess wild-type DNA.

We applied a facile LIF dual-channel monitoring system recently developed and reported by our group to the polymerase chain reaction/ligase detection reaction/CGE method for detecting low-abundance point mutations present in a wild-type sequence-dominated population. Mutation discrimination limits and signaling fidelity of the analytical system were evaluated using three mutant variations in codon 12 of the K-ras oncogene that have high diagnostic value for colorectal cancer. We demonstrated the high sensitivity of the present method by detecting rare mutations present among an excess of wild-type alleles (one mutation among ~100 normal sequences). This method also simultaneously interrogated the allelic compositions of the test samples with high specificity through spectral discrimination of the dye-tagged ligase detection reaction products using the dual-channel monitoring system.

Discriminative detection of low-abundance point mutations using a PCR/ligase detection reaction/capillary gel electrophoresis method and fluorescence dual-channel monitoring.

Chronic expanding haematoma (CEH) is a rare type of haematoma that enlarges slowly and continuously without coagulation. It can occur following surgery because of shear stress-induced bleeding in the scar tissue between the subcutaneous fat and fascia. We present three cases of large chronic CEH that were successfully treated with triamcinolone injections. Three female patients developed large chronic CEH at 9 months, 5 years, and 6 years, respectively, after latissimus dorsi flap harvesting for breast reconstruction. Although the condition did not improve after multiple sessions of haematoma aspiration in the first two patients, it resolved following a single 40-mg triamcinolone injection along with appropriate compression dressing for several weeks. In the third patient, triamcinolone was injected immediately after the initial aspiration of the haematoma, and the condition improved considerably. There were no side effects in any of the patients. To the best of our knowledge, this is the first report of successful treatment of large CEH using triamcinolone. Therefore, we suggest that triamcinolone injections be considered for the treatment of CEH.

Management of chronic expanding haematoma using triamcinolone after latissimus dorsi flap harvesting.

We examined whether brachial-ankle pulse wave velocity (baPWV) and ankle-brachial pressure index (ABI) are predictors for mortality in diabetic patients after lower extremity amputation. This was an observational historical cohort study of 102 Japanese diabetic patients after first non-traumatic lower extremity amputation, with a mean age of 63 years (standard deviation 12 years). The end-point was all-cause mortality. During the mean follow-up period of 3.3 years, 44 patients reached the end-point. In both univariate and multivariate analyses, baPWV (m/s) (hazard ratio [HR] 1.05 and 1.04, both P < 0.01, respectively), but not ABI (HR 0.38 and 0.89, P = 0.08 and 0.86, respectively), was a significant predictor for the end-point. When baPWV (above or below the median [21.8 m/s]) and ABI (normal [0.9-1.4] or not) were analyzed as categorical variables, the results were similar. In conclusion, baPWV, but not ABI, might be a predictor for all-cause mortality in diabetic patients after lower extremity amputation.

Brachial-ankle pulse wave velocity, but not ankle-brachial index, predicts all-cause mortality in patients with diabetes after lower extremity amputation.

Congenital brachymetatarsia most commonly involves the fourth ray and may be combined with metacarpal shortening. Numerous reports have demonstrated the usefulness of gradual lengthening of the metatarsals. However, very few studies have investigated methods of recovering the lost regenerative bone. The patient was a 16-year-old girl with bilateral brachymetatarsia of the fourth metatarsal. After a long consolidation period after gradual lengthening of the fourth metatarsal by 20 mm, the patient had an hourglass-shaped regenerated bone. Therefore, we grafted a bioabsorbable hydroxyapatite and collagen composite. Six months after the surgery, well-regenerated bone could be recognized on radiographic evaluation and was resistant to refracture. Callus distraction is a method that aims to avoid donor site morbidity. The strength of artificial bone is often a problem. Bioabsorbable hydroxyapatite and collagen composite compensates for the lack of regenerated bone and distraction gap and prevents refracture, thus preventing complications after metatarsal distraction.

Treatment of Complications after Distraction Osteogenesis for Brachymetatarsia of the Fourth Metatarsal.

YAP (yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif) are part of a classical pathway that controls contact inhibition in the Hippo pathway. YAP and TAZ were recently reported to act as nuclear relays of mechanical signals that communicate extracellular matrix rigidity and cell shape. However, the role of YAP/TAZ signaling in keloid formation is unclear. Here, we used immunohistochemistry to investigate YAP/TAZ expression in keloid and nonaffected lesions. YAP/TAZ expression in keloid fibroblasts had a greater tendency to localize to the nucleus relative to that seen in fibroblasts from unaffected tissues. Meanwhile, keratinocytes or endothelial cells from either keloid or unaffected tissues showed no significant differences in YAP/TAZ expression patterns. These results suggest that YAP/TAZ nuclear localization in keloid fibroblasts might activate Hippo signaling and may play an important role in gene expression that affects keloid formation and stiffness.

Mariko Hamada

Diabetic Center

Tokyo Women's Medical University School of Medicine

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Mariko Hamada

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Tokyo Women's Medical University School of Medicine

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Diabetic Center