The mitotic kinase Aurora--a promotes distant metastases by inducing epithelial-to-mesenchymal transition in ERα(+) breast cancer cells.

In this study, we demonstrate that constitutive activation of Raf-1 oncogenic signaling induces stabilization and accumulation of Aurora-A mitotic kinase that ultimately drives the transition from an epithelial to a highly invasive mesenchymal phenotype in estrogen receptor α-positive (ERα(+)) breast cancer cells. The transition from an epithelial- to a mesenchymal-like phenotype was characterized by reduced expression of ERα, HER-2/Neu overexpression and loss of CD24 surface receptor (CD24(-/low)). Importantly, expression of key epithelial-to-mesenchymal transition (EMT) markers and upregulation of the stemness gene SOX2 was linked to acquisition of stem cell-like properties such as the ability to form mammospheres in vitro and tumor self-renewal in vivo. Moreover, aberrant Aurora-A kinase activity induced phosphorylation and nuclear translocation of SMAD5, indicating a novel interplay between Aurora-A and SMAD5 signaling pathways in the development of EMT, stemness and ultimately tumor progression. Importantly, pharmacological and molecular inhibition of Aurora-A kinase activity restored a CD24(+) epithelial phenotype that was coupled to ERα expression, downregulation of HER-2/Neu, inhibition of EMT and impaired self-renewal ability, resulting in the suppression of distant metastases. Taken together, our findings show for the first time the causal role of Aurora-A kinase in the activation of EMT pathway responsible for the development of distant metastases in ERα(+) breast cancer cells. Moreover, this study has important translational implications because it highlights the mitotic kinase Aurora-A as a novel promising therapeutic target to selectively eliminate highly invasive cancer cells and improve the disease-free and overall survival of ERα(+) breast cancer patients resistant to conventional endocrine therapy.

Indolactam V/GLP-1-mediated differentiation of human iPS cells into glucose-responsive insulin-secreting progeny.

Nuclear reprogramming of somatic tissue enables derivation of induced pluripotent stem (iPS) cells from an autologous, non-embryonic origin. The purpose of this study was to establish efficient protocols for lineage specification of human iPS cells into functional glucose-responsive, insulin-producing progeny. We generated human iPS cells, which were then guided with recombinant growth factors that mimic the essential signaling for pancreatic development. Reprogrammed with four stemness factors, human fibroblasts were here converted into authentic iPS cells. Under feeder-free conditions, fate specification was initiated with activin A and Wnt3a that triggered engagement into definitive endoderm, followed by priming with fibroblast growth factor 10 (FGF10) and KAAD-cyclopamine. Addition of retinoic acid, boosted by the pancreatic endoderm inducer indolactam V (ILV), yielded pancreatic progenitors expressing pancreatic and duodenal homeobox 1 (PDX1), neurogenin 3 (NGN3) and neurogenic differentiation 1 (NEUROD1) markers. Further guidance, under insulin-like growth factor 1 (IGF-1), hepatocyte growth factor (HGF) and N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT), was enhanced by glucagon-like peptide-1 (GLP-1) to generate islet-like cells that expressed pancreas-specific markers including insulin and glucagon. Derived progeny demonstrated sustained expression of PDX1, and functional responsiveness to glucose challenge secreting up to 230 pM of C-peptide. A pancreatogenic cocktail enriched with ILV/GLP-1 offers a proficient means to specify human iPS cells into glucose-responsive hormone-producing progeny, refining the development of a personalized platform for islet-like cell generation.

Inhibition of HIV-1 replication by simian restriction factors, TRIM5alpha and APOBEC3G.

Old World monkey TRIM5alpha targets incoming human immunodeficiency virus type 1 (HIV-1) viral capsid, whereas the apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like (APOBEC)3 family hypermutate/degrade viral sequences. Here, we show the potentials of simian TRIM5alpha and APOBEC3G as therapeutic sequences for AIDS gene therapy. Both rhesus and African green monkey (agm) TRIM5alpha efficiently restrict HIV-1 vectors with divergent Gag from different HIV-1 subtypes. Human T cells genetically engineered to express agm-TRIM5alpha block or delay HIV-1 replication. Although agm-APOBEC3G expression alone only transiently suppresses HIV-1 replication, co-expression of agm-APOBEC3G and agm-TRIM5alpha successfully block the virus replication for more than 5 weeks.

Hygiene problems of residual limb and silicone liners in transtibial amputees wearing the total surface bearing socket.

OBJECTIVE: To determine whether washing the residual limb and silicone liner reduces the associated skin problems in transtibial amputees who wear a total surface bearing (TSB) socket. DESIGN: Case series. SETTING: General community. PATIENTS: Eighty-three transtibial amputees (65 men, 18 women; mean age, 53.4 yr) in western Japan who used or had used a TSB socket with a silicone liner in the previous 5 years. INTERVENTION: Participants took a self-administered questionnaire that included items concerning their personal profile, daily life activities, period of TSB use (yr), hours of TSB use daily, washing frequency of the residual limb and silicone liner, the method of washing, and any associated hygiene problems. MAIN OUTCOME MEASURES: Logistic regression analysis was applied to examine factors related to hygiene problems. Predicted values include hygiene problems (perspiration, eruptions, itching, odor) and explanatory values include TSB use, daily life activity, and washing of limb and prosthetic. RESULTS: Fifty-five subjects washed the residual limb, and 44 subjects washed the silicone liner every day. Itching, perspiration, eruption, and odor were frequent hygiene problems. Perspiration was noted less by women, eruption more by older subjects, and itching and odor more by younger subjects. Washing the silicone liner every day was associated with fewer reports of skin eruption. CONCLUSION: Keeping the residual limb and silicone liner clean is important to reduce skin problems, but hygiene problems of the residual limb and silicone liner still remain to be resolved.

Moisture permeability of the total surface bearing prosthetic socket with a silicone liner: is it superior to the patella-tendon bearing prosthetic socket?

The purpose of this study was to examine the moisture permeability properties of materials used for total surface bearing (TSB) socket with a silicone liner, a combination of Silicone Suction Socket or Icelandic Roll-On Silicone Socket (ICEROSS) and an acrylic plastic sheet (Degaplast), patella-tendon bearing (PTB) socket, a combination of Pe-Lite and Degaplast, and wooden socket made of poplar. Moisture permeability of the socket materials was measured as the diminution of water in a container after 12 hours in a climatic chamber. Eight containers with their open, top side were uncovered (no material) or sealed with one of the socket materials; the experiment was repeated four times. One-way analysis of variance followed by Bonferroni's test was applied to examine the differences in moisture permeability. Moisture permeability levels were as follows: no material, 85.9 +/- 1.3 g; poplar, 4.3 +/- 0.4 g; Silicone Suction Socket, 1.1 +/- 0.2 g; ICEROSS, 1.0 +/- 0.2 g; Pe-Lite, 0.8 +/- 0.1 g; 3S + Degaplast, 0.8 +/- 0.1 g; ICEROSS + Degaplast, 0.8 +/- 0.2 g; and Pe-Lite + Degaplast, 0.8 +/- 0.1 g. There were significant differences between the uncovered container and the others, and between poplar and the others (P < 0.05). We concluded that the TSB socket with a silicone liner is not superior to the PTB socket with regard to moisture permeability, and that it is necessary to develop a new prosthetic socket that allows heat release and drainage of sweat.

Subjective evaluations and objective measurements of the ischial-ramal containment prosthesis.

We examined 12 transfemoral amputees, 6 using the IRC socket and 6 the QL socket, to confirm whether the ischial-ramal containment (IRC) socket is truly superior to the quadrilateral (QL) socket. In subjective evaluation, the IRC group was significantly better in the total score and in items of comfort, that is, to sit on a chair and lumbar lordosis at heel off (Mann-Whitney test, P < 0.05), better but not significant in the items of comfortable to wear, comfortable to go up and down stairs, and truncal sway during stance phase. By computed tomography, the femur of the IRC group was kept in a position significantly more medial than that of the QL group (Mann-Whitney test, P < 0.05); but no significant difference in gluteal medial muscle atrophy ratios between the two groups was found (Mann-Whitney test, P > 0.05). By X-ray, the stump of the IRC group was maintained significantly more adducted during one foot standing on the prosthesis (Mann-Whitney test, P < 0.05), but the lateral force ratio during mid-stance of the IRC group was smaller, but not significantly, than that of the QL group. Physiological cost index (PCI), an indirect simple method for evaluating oxygen consumption of gait, had no significant difference between the two groups (Mann-Whitney test, P > 0.05), and a multiple regression analysis revealed that the stump length ratio and lateral force ratio during mid-stance were significant explanatory variables for predicting PCI (adjusted R square: 0.87, F-value: 11.85, P < 0.05). The results of this study have revealed that the advantage of the IRC socket is a tender feeling of the stump, but that the metabolic efficiency is not superior to the QL socket at the most comfortable speed.

Total surface bearing below-knee prosthesis: advantages, disadvantages, and clinical implications.

OBJECTIVE: To investigate the total surface bearing (TSB) prosthesis for below-knee amputee patients and to determine its clinical indications. DESIGN: Case series. SETTING: University hospital and the Welfare Center for the Disabled. PATIENTS: Thirty-two subjects who could walk with the TSB prosthesis and had already used the patellar-tendon bearing (or Kondylen-Bettung Münster) socket. MAIN OUTCOME MEASURE: Subjective assessment of the TSB socket, consisting of an overall rating and ratings of 13 individual items, was performed, and results were analyzed with Fisher's exact test. RESULTS: Seventy-five percent of the subjects were satisfied overall with the TSB socket. Of the 13 items, comfort, ease to swing, pain, piston movement, tightness, skin irritation, appearance, and durability were regarded as good by more than 75% of the patients. Donning, perspiration, odor, and staining were regarded as poor by more than 20%. Items that were significantly related with overall satisfaction with the TSB socket were comfort, ease to swing, and piston movement, while donning was significantly related to dissatisfaction. CONCLUSION: The TSB socket is suitable for and preferred by many amputee subjects, but some problems remain to be solved.

Clinical evaluations of dorsiflexion assist controlled by spring ankle-foot orthosis for hemiplegic patients.

To examine the usefulness of Dorsiflexion Assist Controlled by Spring Ankle-Foot Orthoses (DACS-AFO), 5 hemiplegic patients in Brunnstrom's recovery stage IV were prescribed the DACS-AFO, after which their gait with the DACS-AFO was clinically evaluated. Clinical observation did not reveal that their gait with the DACS-AFO was better than that with an AFO which the patients usually used, except when one patient walked on a slope. No significant difference in 10 m walking time was found between when the DACS-AFO and shoehorn type AFO were used. Subjective evaluations by the patients indicated that the DACS-AFO was as good as a shoehorn type AFO in walking on flat and sloped surfaces, though its weight, noise, and appearance caused by the cylinder of the DACS-AFO were regarded as poor. In spite of the excellent biomechanical characteristics of the DACS-AFO, the patients did not adopt it. The subjects in this study had already walked well with their own shoehorn type AFO, and their selection may have been biased toward poor results. However, we did not conclude that the DACS-AFO is superior to the shoehorn type AFO for hemiplegic patients in the stage IV.

Properties of the flexible pressure sensor under laboratory conditions simulating the internal environment of the total surface bearing socket.

The purpose of this study was to investigate the properties of the flexible pressure sensor under laboratory conditions simulating the internal environment of the total surface bearing (TSB) socket to determine optimal conditions for measuring normal stresses on the stump. The equipment used in the study was the Pressure Distribution Sensor System for Sockets. In a climatic chamber maintained at 37 degrees C and 70% humidity the sensor sheet was mounted on a measuring apparatus loaded with three 10 kg weights, and output from the sensor was recorded. Because of sensor creep, a sample 60 seconds after loading was adopted as the measured output. Output was greater when weight was decreased than when weight was increased because of hysteresis (paired t-test, p<0.05). The sensor had temperature sensitivity but differences in output were not statistically significant (paired t-test, 0.10>p>0.05). There were no significant differences in output among five sensor sheets or among five sections of four sensor sheets (two-way ANOVA, p>0.05), but repeated loading on the same section of the sensor sheet increased output (two-way ANOVA, p<0.05). Reproducibility and sensitivity distribution of the sensor are considered satisfactory under laboratory conditions, but measurements of rapid and repetitive movements may not be accurate and comparing subtle changes in output from a single sensor is not suitable. The reliability of the sensor in a clinical setting for measuring normal stresses on the stump with the TSB socket should be examined.