Exosomal proteins as prognostic biomarkers in non-small cell lung cancer.

BACKGROUND: Use of exosomes as biomarkers in non-small cell lung cancer (NSCLC) is an intriguing approach in the liquid-biopsy era. Exosomes are nano-sized vesicles with membrane-bound proteins that reflect their originating cell. Prognostic biomarkers are needed to improve patient selection for optimal treatment. We here evaluate exosomes by protein phenotyping as a prognostic biomarker in NSCLC. METHODS: Exosomes from plasma of 276 NSCLC patients were phenotyped using the Extracellular Vesicle Array; 49 antibodies captured the proteins on the exosomes, and a cocktail of biotin-conjugated antibodies binding the general exosome markers CD9, CD81 and CD63 was used to visualise the captured exosomes. For each individual membrane-bound protein, results were analysed based on presence, in a concentration-dependent manner, and correlated to overall survival (OS). RESULTS: The 49 proteins attached to the exosomal membrane were evaluated. NY-ESO-1, EGFR, PLAP, EpCam and Alix had a significant concentration-dependent impact on inferior OS. Due to multiple testing, NY-ESO-1 was the only marker that maintained a significant impact on inferior survival (hazard rate (HR) 1.78 95% (1.78-2.44); p = 0.0001) after Bonferroni correction. Results were adjusted for clinico-pathological characteristics, stage, histology, age, sex and performance status. CONCLUSION: We illustrate the promising aspects associated with the use of exosomal membrane-bound proteins as a biomarker and demonstrate that they are a strong prognostic biomarker in NSCLC.

Does smoking, age or gender affect the protein phenotype of extracellular vesicles in plasma?

Extracellular vesicles (EVs) are involved in several diseases, which have formed the basis for the potential use of EV analyses in a clinical setting. The protein phenotype of EVs can provide information on the functionality of the vesicles and may be used for identification of disease-related biomarkers. With this extensive study of 161 healthy individuals it was elucidated that certain markers of plasma EVs are influenced by demographic variations such as gender, age and smoking status. When the purpose is to use EVs as a diagnostic tool, it should be emphasized how important it is to choose the correct demographic group when comparing marker levels of plasma EVs.

Prolongation of the rat composite tissue allograft survival by the combination of tolerogenic immature dendritic cells and short-term treatment with FK506.

OBJECTIVE: Prevention of rejection in composite tissue allotransplantation without continuous immunosuppression is of paramount importance in the field of transplantation. Recently dendritic cells (DCs) have gained considerable attention as antigen-presenting cells that are also capable of tolerance induction. This study assessed the effect of interleukin (IL)-10-supplemented, alloantigen-pulsed immature tolerogenic DC to increase survival of orthotopic hind limb transplantations in rats. MATERIALS AND METHODS: Hind limbs from Sprague-Dawley (SD) hosts were transplanted to Fischer 344 (F344) rats. Peripheral blood mononuclear cells (PBMC) isolated from F344 were cultured to generate immature DCs (imDCs); IL-10 was added for tolerogenic DC induction. Flow cytometric analysis were performed to characterize the DC phenotype. IL-10-imDCs cocultured with donor mononuclear cells for 24 hours were reinjected into recipients subcutaneously 1 day before transplantation. Recipient animals were divided into 4 groups, each comprising 6 rats (n = 6): group I (untreated controls), group II (IL-10-imDCs alone), group III (FK-506 [Tacrolimus, 2 mg/kg] for 2 weeks postoperative), and group IV (recipient origin IL-10-imDCs combined with FK-506 for 2 weeks postoperative). Observation of graft appearance, cytokine production assays, and confocal immunofluorescence were performed at postoperative 1 week. RESULTS: The combination of IL-10-treated imDCs and FK-506 produced a significantly prolonged median allograft survival (46.7 days) compared with groups I (4.7), II (5.3), or III (26.3). Splenocytes isolated from rats treated with IL-10-imDCs plus and FK-506 produced significant amounts of IL-10 and IL-4 cytokines upon alloantigen stimulation, as confirmed using ELISPOT and ELISA. CONCLUSIONS: We demonstrated that IL-10-treated imDCs induced T-cell hyporesponsiveness, skewing the immune response to Th2 elements, resulting in long-term survival of composite tissue allografts.

Unilateral buttock reconstruction using contralateral inferior gluteal artery perforator flap with the aid of multi-detector CT.

SUMMARY: The applicability of inferior gluteal artery perforator (IGAP) flaps in breast reconstruction following breast cancer has been well described. However, the use of IGAP flaps in buttock augmentation has not been presented. We present the case of a female patient with buttock asymmetry and a deficiency of volume, who underwent buttock reconstruction based on a contralateral IGAP flap. Multidetector computed tomography (MDCT) was used to study donor and recipient areas, and allowed easy interpretation as it provided anatomical images and three-dimensional anatomy reconstructions. Based on a knowledge of individual anatomical perforating vessel distributions, safe perforator flaps can be designed. Moreover, we are convinced that buttock reconstruction using autologous tissue can be performed in a safe and reliable fashion using perforator flaps. Donor site morbidity was minimal and the muscle at the donor site was preserved. The contralateral buttock proved an excellent donor site for aesthetic unilateral buttock reconstruction and provided ample tissue in the described case.

Complete correction of enzymatic deficiency and neurochemistry in the GM1-gangliosidosis mouse brain by neonatal adeno-associated virus-mediated gene delivery.

GM1-gangliosidosis is a glycosphingolipid (GSL) lysosomal storage disease caused by autosomal recessive deficiency of lysosomal acid beta-galactosidase (betagal), and characterized by accumulation of GM1-ganglioside and GA1 in the brain. Here we examined the effect of neonatal intracerebroventricular (i.c.v.) injection of an adeno-associated virus (AAV) vector encoding mouse betagal on enzyme activity and brain GSL content in GM1-gangliosidosis (betagal(-/-)) mice. Histological analysis of betagal distribution in 3-month-old AAV-treated betagal(-/-) mice showed that enzyme was present at high levels throughout the brain. Biochemical quantification showed that betagal activity in AAV-treated brains was 7- to 65-fold higher than in wild-type controls and that brain GSL levels were normalized. Cerebrosides and sulfatides, which were reduced in untreated betagal(-/-) mice, were restored to normal levels by AAV treatment. In untreated betagal(-/-) brains, cholesterol was present at normal levels but showed abnormal cellular distribution consistent with endosomal/lysosomal localization. This feature was also corrected in AAV-treated mice. The biochemical and histological parameters analyzed in this study showed that normal brain neurochemistry was achieved in AAV-treated betagal(-/-) mice. Therefore we show for the first time that neonatal AAV-mediated gene delivery of lysosomal betagal to the brain may be an effective approach for treatment of GM1-gangliosidosis.

A new surgical treatment of keloid: keloid core excision.

Keloids and hypertrophic scars result from excessive collagen deposition, the cause of which is not yet known. Unlike hypertrophic scars, keloids frequently persist at the site of injury, often recur after excision and always overgrow the boundaries of the original wound. There have been many trials to control keloids, but most of them have been unsuccessful. The authors propose a new surgical technique to treat keloids and name it keloid core extirpation. They excise the inner fibrous core from the keloid and cover the defect with a keloid rind flap, which is arterialized by the subcapsular vascular plexus. The authors treated 24 keloids of the ear, trunk, face, and genitalia with keloid core excision. Four cases of partial rind flap congestion or necrosis occurred. Those patients who healed primarily after surgery showed no evidence of keloid recurrence as long as they were followed. The authors have found the keloid core extirpation technique to be excellent in preventing keloid recurrence, with no adjuvant therapy after surgery.

Refinement in aesthetic contouring of the prominent mandibular angle.

Oriental women, in general, greatly desire a more delicate and feminine facial shape. This can be obtained by contouring the prominent mandibular angles that give a strong, masculine image. Western authors regarded masseteric muscular hypertrophy the main cause of a square facial appearance, so they usually corrected it by partially excising the masseter muscle. In the authors' view, a square facial appearance in the Oriental is not due to masseteric hypertrophy but to a posterior projection and lateral flaring of the mandibular angle. However, it is sometimes difficult to make the square face narrow and ovoid by using only the conventional curved-angle osteotomy. We divided patients, whose chief complaint was a square facial appearance, into three groups after clinical, photographic, and radiographic evaluation. We applied different contouring methods to each of the three groups and obtained cosmetically improved facial appearance in both lateral and frontal views.

Profiloplasty of the lower face by maxillary and mandibular anterior segmental osteotomies.

Maxillary and mandibular anterior segmental osteotomies (ASO) are ways to correct disharmony in the lip (contour, lip seal, and profile) and occasional dentoalveolar malocclusion. We performed 23 maxillary setback ASO, three maxillary advancement ASO, 21 mandibular setback ASO, and six mandibular advancement ASO in 28 patients to improve their lower facial profile. Other combined operative procedures include nine LeFort 1 osteotomies, four bilateral mandibular ramus osteotomies, two genioplasties, four mandibular angle contouring procedures, two reduction malar plasties, two piriform augmentations with bone graft, and a facelift for correcting of long faces, asymmetric faces, severe malocclusions, microgenias, prominent mandibular angles, prominent malar eminences, piriform recessions, and an aging face. Twenty five patients were satisfied with the results. Two patients complained of an over-recessed lip region, and one of septal deviation. There were no other significant complications or relapses throughout the followup period (9-30 months). Maxillary and mandibular ASO are effective, selective, relatively safe, and simple methods for correcting lower facial profile disharmony to attain a satisfactory aesthetic facial contour.