Circulating Levels of MiRNAs From 320 Family in Subjects With Lipodystrophy: Disclosing Novel Signatures of the Disease.

Lipodystrophy (LD) indicates a group of rare disorders, with generalized or partial loss of white adipose tissue (WAT) often associated with metabolic derangements. Heterogeneity/wide spectrum of the disease and lack of biomarkers make diagnosis often difficult. MicroRNAs are important to maintain a correct WAT function and WAT is a source of circulating miRNAs (cmiRs). miRNAs from 320 family were previously detected in the WAT and variably associated to the metabolic syndrome. Our aim was then to investigate if LD can result in altered abundance of cmiRs-320. We collected samples from a cohort of LD subjects of various subtypes and from age matched controls. Use of quantitative PCR determined that cmiRs- 320a-3p, 320b, 320c, 320e are upregulated, while 320d is downregulated in LD. CmiRs-320 power as classifiers was more powerful in the most extreme and defined forms of LD, including the generalized and the Dunnigan subtypes. cmiR-320a-3p showed significant inverse relationships with plasma leptin (P < 0.0001), typically low in LD. The hepatic enzymes gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and the marker of inflammation C-reactive protein (CRP) were inversely related to cmiR 320d (P < 0.05, for CRP and GGT; P < 0.01, for AST and ALT). Gene ontology analysis revealed cell-cell adhesion as a process regulated by 320 miRNAs targets, thus disclosing a novel route to investigate origin of WAT loss/dysfunction. In conclusion, cmiRs-320 constitute novel biomarkers of LD, abundance of miR320a-3p is inversely associated to indicators related to WAT function, while downregulation of cmiR-320d predicts an altered hepatic profile and higher inflammation.

Interactive design of patient-oriented video-games for rehabilitation: concept and application.

PURPOSE: Serious video-games are innovative tools used to train the motor skills of subjects affected by neurological disorders. They are often developed to train a specific type of patients and the rules of the game are standardly defined. A system that allows the therapist to design highly patient-oriented video-games, without specific informatics skills, is proposed. METHOD: The system consists of one personal computer, two screens, a Kinect™ sensor and a specific software developed here for the design of the video-games. It was tested with the collaboration of three therapists and six patients, and two questionnaires were filled in by each patient to evaluate the appreciation of the rehabilitative sessions. RESULTS: The therapists learned easily how to use the system, and no serious difficulties were encountered by the patients. The questionnaires showed an overall good satisfaction by the patients and highlighted the key-role of the therapist in involving the patients during the rehabilitative session. CONCLUSIONS: It was found that the proposed system is effective for developing patient-oriented video-games for rehabilitation. The two main advantages are that the therapist is allowed to (i) develop personalized video-games without informatics skills and (ii) adapt the game settings to patients affected by different pathologies. Implications for rehabilitation Virtual reality and serious video games offer the opportunity to transform the traditional therapy into a more pleasant experience, allowing patients to train their motor and cognitive skills. Both the therapists and the patients should be involved in the development of rehabilitative solutions to be highly patient-oriented. A system for the design of rehabilitative games by the therapist is described and the feedback of three therapists and six patients is reported.

Conceptual design and implantation of an external fixator with improved mobility for knee rehabilitation.

A hinged external fixator is used to allow early knee rehabilitation in case of injury or trauma, as an alternative approach to immobilization. It is mainly adopted for the treatment of dislocations, which involve tearing of the ligaments, and it basically consists of two links connected to each other by a revolute joint. Each link is fixed to the femur and tibia via pin fixation, and the revolute joint is approximately aligned to the knee flexion-extension (FE) axis. The advantage in its implantation is to protect ligament reconstruction, while allowing for an aggressive rehabilitation. Traditional fixators only accommodate the functional flexion movement in a limited range, i.e. where the anatomical movement is closer to a planar circular trajectory. This paper presents the conceptual design and implantation procedure of a double-axis fixator, which accommodates both FE and longitudinal internal-external rotation. The procedure is based on accurate knee kinematics measurements and on computer-aided multibody simulations to assist clinicians in the implantation. An experimental test is presented using an artificial knee, and guidelines are given for in vitro studies. The proposed technique may allow for a better understanding of knee kinematics and have the potential advantage to increase the range of motion in postoperative rehabilitation.

On the estimate of the two dominant axes of the knee using an instrumented spatial linkage.

This article presents the validation of a technique to assess the appropriateness of a 2 degree-of-freedom model for the human knee, and, in which case, the dominant axes of flexion/extension and internal/external longitudinal rotation are estimated. The technique relies on the use of an instrumented spatial linkage for the accurate detection of passive knee kinematics, and it is based on the assumption that points on the longitudinal rotation axis describe nearly circular and planar trajectories, whereas the flexion/extension axis is perpendicular to those trajectories through their centers of rotation. By manually enforcing a tibia rotation while bending the knee in flexion, a standard optimization algorithm is used to estimate the approximate axis of longitudinal rotation, and the axis of flexion is estimated consequently. The proposed technique is validated through simulated data and experimentally applied on a 2 degree-of-freedom mechanical joint. A procedure is proposed to verify the fixed axes assumption for the knee model. The suggested methodology could be possibly valuable in understanding knee kinematics, and in particular for the design and implant of customized hinged external fixators, which have shown to be effective in knee dislocation treatment and rehabilitation.

Validation of a calibration technique for 6-DOF instrumented spatial linkages.

This paper presents a practical and effective approach to the calibration of instrumented spatial linkages for biomechanical applications. A 6-DOF mechanical linkage with rotational transducers is designed and in-house manufactured for this purpose. In order to assess the validity of the proposed calibration technique and to distinguish between geometrical and electrical parameters uncertainties, high-precision optical encoders are used and calibration is addressed from a kinematic point of view only. The proposed technique is based on a closed-loop method, in which the end segments of the linkage are connected to each other by revolute joints. A parametrical model of the system is formulated using a standard link-to-link transformation matrices approach. Continuous data collection is carried out and a recursive identification of kinematic parameters is implemented by the use of an extended Kalman filter algorithm. Results shows that the proposed technique, despites its simplicity, is effective in improving the accuracy of the system up to its theoretically computed resolution, which limits the performance of the real system.

Genetic analysis of TTF-2 gene in children with congenital hypothyroidism and cleft palate, congenital hypothyroidism, or isolated cleft palate.

Homozygous null mice for thyroid transcription factor (TTF)-2 gene exhibit cleft palate and thyroid malformation. We performed a genetic analysis of the TTF-2 gene in 2 children with congenital hypothyroidism (CH) and cleft palate, 45 children with thyroid dysgenesis, 19 children with isolated cleft palate or cleft lip, 4 patients with thyroid hemiagenesis. The entire coding-region of the TTF-2 gene was analyzed by direct sequencing. Direct sequencing of the TTF-2 gene revealed polymorphisms in the length of the polyalanine tract. The most frequent stretch length was 14 residues and it was found in 50 of 70 (71%) and in 45 of 53 (85%) normal healthy controls. A polyalanine tract of 16 residues in the heterozygous state was seen in 18 of 70 (26%) cases and in 4 of 53 (7%) normal subjects. In 1 of 4 (25%) case of hemiagenesis a polyalanine tract of 16 residues in the homozygous state was observed. In 1 of 26 agenesis the polyalanine tract consisted of 12 residues in the heterozygous state. Direct sequencing also revealed the presence of two silent polymorphisms. No mutations were identified in the TTF-2 gene. In conclusion, our results show that no genetic alteration was present in the TTF-2 gene of these patients, suggesting that defects in the TTF-2 gene are a rare event.

Role for inner ring deiodination preventing transcutaneous passage of thyroxine.

Creams containing thyroid hormone are commonly employed for cosmetic purposes. To verify whether T(4) applied to the skin surface can enter the bloodstream either directly or as a metabolite, a cream containing L-T(4) [3,5,3',5'-tetraiodothyronine (T(4))] was self-applied by volunteers for 2 wk. No significant variations in urinary iodide, TSH, and serum (total and free) T(4) and T(3) concentrations were observed at any time relative to pretreatment values, whereas rT(3) concentrations increased significantly 6 and 12 h after cream application. The increased rT(3) concentration led us to investigate the presence of inner ring type III deiodinase (D3) activity in human skin. Using human surgical discard skin, we found that T(4) can be carried across human epidermis in a liposome cream. Substantial inner ring deiodination was suggested by the fact that only 10% of transferred thyroid hormone remained as T(4), and T(3) was not detected. We then measured D3 activity in a surgical skin specimen. The K(m) for T(3) was 1.74 nmol/liter, and the maximum velocity was 23.5 fmol/microg microsomal protein/h. In conclusion, our study indicates that normal human skin serves as a substantial, but incomplete, barrier to T(4) passage. D3 plays an important role in augmenting T(4) blockade by inactivating T(4) to rT(3).