Simultaneous measurements of kinematics and fMRI: compatibility assessment and case report on recovery evaluation of one stroke patient

<p>Abstract</p> <p>Background</p> <p>Correlating the features of the actual executed movement with the associated cortical activations can enhance the reliability of the functional Magnetic Resonance Imaging (fMRI) data interpretation. This is crucial for longitudinal evaluation of motor recovery in neurological patients and for investigating detailed mutual interactions between activation maps and movement parameters.</p> <p>Therefore, we have explored a new set-up combining fMRI with an optoelectronic motion capture system, which provides a multi-parameter quantification of the performed motor task.</p> <p>Methods</p> <p>The cameras of the motion system were mounted inside the MR room and passive markers were placed on the subject skin, without any risk or encumbrance. The versatile set-up allows 3-dimensional multi-segment acquisitions including recording of possible mirror movements, and it guarantees a high inter-sessions repeatability.</p> <p>We demonstrated the integrated set-up reliability through compatibility tests. Then, an fMRI block-design protocol combined with kinematic recordings was tested on a healthy volunteer performing finger tapping and ankle dorsal- plantar-flexion. A preliminary assessment of clinical applicability and perspectives was carried out by pre- and post rehabilitation acquisitions on a hemiparetic patient performing ankle dorsal- plantar-flexion. For all sessions, the proposed method integrating kinematic data into the model design was compared with the standard analysis.</p> <p>Results</p> <p>Phantom acquisitions demonstrated the not-compromised image quality. Healthy subject sessions showed the protocols feasibility and the model reliability with the kinematic regressor. The patient results showed that brain activation maps were more consistent when the images analysis included in the regression model, besides the stimuli, the kinematic regressor quantifying the actual executed movement (movement timing and amplitude), proving a significant model improvement. Moreover, concerning motor recovery evaluation, after one rehabilitation month, a greater cortical area was activated during exercise, in contrast to the usual focalization associated with functional recovery. Indeed, the availability of kinematics data allows to correlate this wider area with a higher frequency and a larger amplitude of movement.</p> <p>Conclusions</p> <p>The kinematic acquisitions resulted to be reliable and versatile to enrich the fMRI images information and therefore the evaluation of motor recovery in neurological patients where large differences between required and performed motion can be expected.</p

Biomarker characterization in endometrial cancer in Europe: first survey data analysis from 69 pathological academic and hospital labs

Introduction. Endometrial carcinoma (EC) is the commonest gynecological cancer affecting women in Western populations. To predict patient risk, the 2020 edition of the World Health Organization (WHO) Classification of Tumors of the Female Genital Tract stressed the importance of integrated histo-molecular classification of the disease. This survey analysis poses attention on the most frequently used immunohistochemical and molecular markers adopted in daily categorization of ECs in European laboratories. Methods. We analyzed data collected through questionnaires administered to 40 Italian, 20 Spanish, 3 Swiss and 6 United Kingdom (UK) laboratories. We collected information regarding daily practice in EC evaluation, specifically concerning mismatch repair status (MMR) and microsatellite instability (MSI). Summary and descriptive statistical analyses were carried out to evaluate the current practice of each laboratory. Results. The results show that MMR status is mainly evaluated by using immunohistochemistry (IHC) on most EC samples. The most frequent approach for the analysis of MMR status is IHC of four proteins (PMS2, MSH6, MSH2, MLH1). MSI analysis by molecular methods is uncommon but useful as a supplemental tool in specific conditions. MLH1 promoter hypermethylation and BRAF V600 mutations analysis are performed in case of negative expression of MLH1/PMS2. Other markers (mainly p53 followed by POLE and PTEN) are investigated in particular in Spain and Switzerland in a consistent number of cases. Conclusion. Guidelines consultation and standardization of laboratory procedures are efficient means for EC prognostic risk stratification and improving the quality of care

Re-thinking the role of motor cortex: Context-sensitive motor outputs?

The standard account of motor control considers descending outputs from primary motor cortex (M1) as motor commands and efference copy. This account has been challenged recently by an alternative formulation in terms of active inference: M1 is considered as part of a sensorimotor hierarchy providing top-down proprioceptive predictions. The key difference between these accounts is that predictions are sensitive to the current proprioceptive context, whereas efference copy is not. Using functional electric stimulation to experimentally manipulate proprioception during voluntary movement in healthy human subjects, we assessed the evidence for context sensitive output from M1. Dynamic causal modeling of functional magnetic resonance imaging responses showed that FES altered proprioception increased the influence of M1 on primary somatosensory cortex (S1). These results disambiguate competing accounts of motor control, provide some insight into the synaptic mechanisms of sensory attenuation and may speak to potential mechanisms of action of FES in promoting motor learning in neurorehabilitation

Post-operative acute urinary retention after greenlight laser. Analysis of risk factors from a multicentric database

Purpose: Greenlight laser is a mini-invasive technique used to treat Benign Prostatic Obstruction (BPO). Some of the advantages of GreenLight photoselective vaporization (PVP) are shorter catheterization time and hospital stay compared to TURP. Post-operative acute urinary retention (pAUR) leads to patients' discomfort, prolonged hospital stay and increased health care costs. We analyzed risk factors for urinary retention after GreenLight laser PVP. Materials and methods: In a multicenter experience, we retrospectively analyzed the onset of early and late post-operative acute urinary retention in patients undergoing standard or anatomical PVP. The pre-, intra- and post-operative characteristics were compared betweene patients who started to void and the patients who developed post-operative urinary retention. Results: The study included 434 patients suitable for the study. Post-operative acute urinary retention occurred in 39 (9%). Patients with a lower prostate volume (P &lt; .001), an adenoma volume lower than 40 mL (P &lt; .001), and lower lasing time (P = .013) had a higher probability to develop pAUR at the univariate analysis. The multivariate logistic regression confirmed that lower lasing time (95% CI: 0.86-0.99, OR = 0.93, P = .046) and adenoma volume (95% CI: 0.89-0.98, OR = 0.94, P = .006) are correlated to pAUR. Furthermore IPSS ≥ 19 (95% CI: 1.19- 10.75, OR = 2.27, P = .023) and treatment with 5-ARI (95% CI: 1.05-15.03, OR = 3.98, P = .042) are risk factors for pAUR. Conclusion: In our series, post-operative acute urinary retention was related to low adenoma volume and lasing time, pre-operative IPSS ≥ 19 and 5-ARI intake. These data should be considered in deciding the best timing for urethral catheters removal

La confiance mutuelle dans l'espace pénal européen/Mutual Trust in the European Criminal Area

L'ouvrage constitue une réflexion approfondie sur la problématique de la confiance mutuelle dans l'espace pénal européen, un thème d’une brûlante actualité dès lors que l'importance de la confiance mutuelle dans cet espace va croissant

Legal migration and the fight against illegal immigration

This is the archive of a lecture given by Franco Frattini, Vice-President of the European Commission and Commissioner for Freedom, Security, and Justice. Moderator: Elizabeth Prodromou, Assistant Professor of International Relations, Boston University. This lecture originally aired on WBUR's World of Ideas.European Studies (Boston University; European Studies (Harvard University

National diplomacies and the EEAS

This contribution was delivered on the occasion of the EUI State of the Union in Florence on 9 May 2011

Amino acid oxidases in red biotechnologies: a target and a tool.

D-Amino acid oxidase (DAAO; EC 1.4.3.3) has been proposed to play a main role in the degradation of D-serine, an allosteric activator of the N-methyl-D-aspartate-type glutamate receptor in the human brain, associated with the onset of schizophrenia. To prevent excessive D-serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. The properties of rat DAAO (rDAAO) are unknown and various in vivo trials reported on the effects of DAAO inhibitors on D-serine concentration in rats. rDAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase-oxidase class of flavoproteins. rDAAO differs significantly from the human enzyme because it: 1)) possesses a different substrate specificity; 2) shows a lower kinetic efficiency (because of a low substrate affinity); 3) differs in affinity for binding of classical inhibitors; 4) is a stable monomer; 5) interacts with the mammalian protein modulator pLG72 yielding a \uf07e 100 kDa complex in addition to the \uf07e 200 kDa, as formed by the human DAAO. Interestingly, the concentration of endogenous D-serine in U87 glioblastoma cells was not affected by transfection with rDAAO whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of rat as model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans. During past years, a number of variants of D-amino acid oxidase from the yeast Rhodotorula gracilis (RgDAAO) with altered substrate specificity (e.g., active on acidic, or hydrophobic, or on all D-amino acids) both by rational design and directed evolution methods have been produced in our laboratory. RgDAAO is the most suitable biotechnological tool for the detection of D-amino acids and in this work we evaluated the capability of some mutant forms of this flavoenzyme previously produced in our laboratory in order to determine D-amino acid content in different biological samples. The kinetic constants for a number of natural and unnatural D-amino acids have been investigated. This information constitutes the basis for considering potential analytical applications of these variants of RgDAAO. Glycine is implicated in several physiological functions, e.g. as a biosynthetic precursor or neurotransmitter in the central nervous system. Glycine is an important coagonist of NMDA receptor and it is putatively involved in schizophrenia susceptibility and other neurological diseases, such as congenital nonketotic hyperglycinemia. With the final aim to produce an optimized enzyme that can be employed in a specific biosensor for glycine detection, glycine oxidase from Bacillus subtilis (GO) was engineered to improve its kinetic efficiency on this small amino acid. Based on in silico analysis, site saturation mutagenesis was independently performed at positions Met49, Gly51, Ala54, Met95, Tyr241, His244, Tyr246, Met261, Arg302, Arg329 and Asn330. The GO variants were screened by employing a rapid colorimetric assay on 96 well-plates based on the determination of hydrogen peroxide produced on glycine as substrate: seven GO variants were selected. Significant alteration of kinetic parameters was observed for H244K and H244R GO variants: kcat increased about twice and Km decreased 3-5-fold, yielding a 7-12-fold higher kinetic efficiency on glycine, as compared to the wild-type GO. Screening of GO variants at position 49 also identified an improved enzyme (M49I) showing a 1.4-fold decreased Km. Combination of information gathered from the site saturation mutagenesis approach could be useful to obtain an evolved GO variant suitable for biotechnological applications

Amino acid oxidases in red biotechnologies: a target and a tool.

D-Amino acid oxidase (DAAO; EC 1.4.3.3) has been proposed to play a main role in the degradation of D-serine, an allosteric activator of the N-methyl-D-aspartate-type glutamate receptor in the human brain, associated with the onset of schizophrenia. To prevent excessive D-serine degradation, novel drugs for schizophrenia treatment based on DAAO inhibition were designed and tested on rats. The properties of rat DAAO (rDAAO) are unknown and various in vivo trials reported on the effects of DAAO inhibitors on D-serine concentration in rats. rDAAO was efficiently expressed in Escherichia coli. The recombinant enzyme was purified as an active, 40 kDa monomeric flavoenzyme showing the basic properties of the dehydrogenase-oxidase class of flavoproteins. rDAAO differs significantly from the human enzyme because it: 1)) possesses a different substrate specificity; 2) shows a lower kinetic efficiency (because of a low substrate affinity); 3) differs in affinity for binding of classical inhibitors; 4) is a stable monomer; 5) interacts with the mammalian protein modulator pLG72 yielding a 100 kDa complex in addition to the 200 kDa, as formed by the human DAAO. Interestingly, the concentration of endogenous D-serine in U87 glioblastoma cells was not affected by transfection with rDAAO whereas it was significantly decreased when expressing the human homologue. These results raise doubt on the use of rat as model system for testing new drugs against schizophrenia and indicate a different physiological function of DAAO in rodents and humans. During past years, a number of variants of D-amino acid oxidase from the yeast Rhodotorula gracilis (RgDAAO) with altered substrate specificity (e.g., active on acidic, or hydrophobic, or on all D-amino acids) both by rational design and directed evolution methods have been produced in our laboratory. RgDAAO is the most suitable biotechnological tool for the detection of D-amino acids and in this work we evaluated the capability of some mutant forms of this flavoenzyme previously produced in our laboratory in order to determine D-amino acid content in different biological samples. The kinetic constants for a number of natural and unnatural D-amino acids have been investigated. This information constitutes the basis for considering potential analytical applications of these variants of RgDAAO. Glycine is implicated in several physiological functions, e.g. as a biosynthetic precursor or neurotransmitter in the central nervous system. Glycine is an important coagonist of NMDA receptor and it is putatively involved in schizophrenia susceptibility and other neurological diseases, such as congenital nonketotic hyperglycinemia. With the final aim to produce an optimized enzyme that can be employed in a specific biosensor for glycine detection, glycine oxidase from Bacillus subtilis (GO) was engineered to improve its kinetic efficiency on this small amino acid. Based on in silico analysis, site saturation mutagenesis was independently performed at positions Met49, Gly51, Ala54, Met95, Tyr241, His244, Tyr246, Met261, Arg302, Arg329 and Asn330. The GO variants were screened by employing a rapid colorimetric assay on 96 well-plates based on the determination of hydrogen peroxide produced on glycine as substrate: seven GO variants were selected. Significant alteration of kinetic parameters was observed for H244K and H244R GO variants: kcat increased about twice and Km decreased 3-5-fold, yielding a 7-12-fold higher kinetic efficiency on glycine, as compared to the wild-type GO. Screening of GO variants at position 49 also identified an improved enzyme (M49I) showing a 1.4-fold decreased Km. Combination of information gathered from the site saturation mutagenesis approach could be useful to obtain an evolved GO variant suitable for biotechnological applications