Organoids Modeling Stroke in a Petri Dish.

Stroke is a common neurological disorder, the second leading cause of death, and the third leading cause of disability. Unfortunately, the only approved drug for it is tissue plasminogen, but the therapeutic window is limited. In this context, preclinical studies are relevant to better dissect the underlying mechanisms of stroke and for the drug screening of potential therapies. Brain organoids could be relevant in this setting. They are derived from pluripotent stem cells or isolated organ progenitors that differentiate to form an organ-like tissue, exhibiting multiple cell types that self-organize to form a structure not unlike the organ in vivo. Brain organoids mimic many key features of early human brain development at molecular, cellular, structural, and functional levels and have emerged as novel model systems that can be used to investigate human brain diseases including stroke. Brain organoids are a promising and powerful tool for ischemic stroke studies; however, there are a few concerns that need to be addressed, including the lack of vascularization and the many cell types that are typically present in the human brain. The aim of this review is to discuss the potential of brain organoids as a novel model system for studying ischemic stroke, highlighting both the advantages and disadvantages in the use of this technology.

Brain Organoids: A Game-Changer for Drug Testing.

Neurological disorders are the second cause of death and the leading cause of disability worldwide. Unfortunately, no cure exists for these disorders, but the actual therapies are only able to ameliorate people's quality of life. Thus, there is an urgent need to test potential therapeutic approaches. Brain organoids are a possible valuable tool in the study of the brain, due to their ability to reproduce different brain regions and maturation stages; they can be used also as a tool for disease modelling and target identification of neurological disorders. Recently, brain organoids have been used in drug-screening processes, even if there are several limitations to overcome. This review focuses on the description of brain organoid development and drug-screening processes, discussing the advantages, challenges, and limitations of the use of organoids in modeling neurological diseases. We also highlighted the potential of testing novel therapeutic approaches. Finally, we examine the challenges and future directions to improve the drug-screening process.

The spatio-temporal architecture of everyday manual behavior.

In everyday activities, humans move alike to manipulate objects. Prior works suggest that hand movements are built by a limited set of basic building blocks consisting of a set of common postures. However, how the low dimensionality of hand movements supports the adaptability and flexibility of natural behavior is unknown. Through a sensorized glove, we collected kinematics data from thirty-six participants preparing and having breakfast in naturalistic conditions. By means of an unbiased analysis, we identified a repertoire of hand states. Then, we tracked their transitions over time. We found that manual behavior can be described in space through a complex organization of basic configurations. These, even in an unconstrained experiment, recurred across subjects. A specific temporal structure, highly consistent within the sample, seems to integrate such identified hand shapes to realize skilled movements. These findings suggest that the simplification of the motor commands unravels in the temporal dimension more than in the spatial one.

Early EEG responses to pre-electoral survey items reflect political attitudes and predict voting behavior.

Self-reports are conventionally used to measure political preferences, yet individuals may be unable or unwilling to report their political attitudes. Here, in 69 participants we compared implicit and explicit methods of political attitude assessment and focused our investigation on populist attitudes. Ahead of the 2019 European Parliament election, we recorded electroencephalography (EEG) from future voters while they completed a survey that measured levels of agreement on different political issues. An Implicit Association Test (IAT) was administered at the end of the recording session. Neural signals differed as a function of future vote for a populist or mainstream party and of whether survey items expressed populist or non-populist views. The combination of EEG responses and self-reported preferences predicted electoral choice better than traditional socio-demographic and ideological variables, while IAT scores were not a significant predictor. These findings suggest that measurements of brain activity can refine the assessment of socio-political attitudes, even when those attitudes are not based on traditional ideological divides.

Inkjet-printed fully customizable and low-cost electrodes matrix for gesture recognition.

The use of surface electromyography (sEMG) is rapidly spreading, from robotic prostheses and muscle computer interfaces to rehabilitation devices controlled by residual muscular activities. In this context, sEMG-based gesture recognition plays an enabling role in controlling prosthetics and devices in real-life settings. Our work aimed at developing a low-cost, print-and-play platform to acquire and analyse sEMG signals that can be arranged in a fully customized way, depending on the application and the users' needs. We produced 8-channel sEMG matrices to measure the muscular activity of the forearm using innovative nanoparticle-based inks to print the sensors embedded into each matrix using a commercial inkjet printer. Then, we acquired the multi-channel sEMG data from 12 participants while repeatedly performing twelve standard finger movements (six extensions and six flexions). Our results showed that inkjet printing-based sEMG signals ensured significant similarity values across repetitions in every participant, a large enough difference between movements (dissimilarity index above 0.2), and an overall classification accuracy of 93-95% for flexion and extension, respectively.

Effects of cognitive workload on heart and locomotor rhythms coupling.

Different physiological signals could be coupled under specific conditions, in some cases related to pathologies or reductions in system complexity. Cardiac-locomotor synchronization (CLS) has been one of the most investigating coupling. The influence of a cognitive task on walking was investigated in dual-task experiments, but how different cognitive tasks may influence CLS has poorly been investigated. Twenty healthy subjects performed a dual-task walking (coupled with verbal fluency vs calculation) on a treadmill at three different speeds (comfortable speed CS; fast-speed: CS + 2 km/h; slow-speed: CS-2 km/h) while cardiac and walking rhythms were recorded using surface electrodes and a triaxial accelerometer, respectively. According to previous studies, we found a cognitive-motor interference for which cognitive performance was affected by motor exercise, but not vice-versa. We found a CLS at the baseline condition, at fast speed in both cognitive tasks, while at comfortable speed only for the verbal fluency task. In conclusion, the cardiac and locomotor rhythms were not coupled at slow speed and at comfortable speed during subtraction task. Cognitive performances generally increased at faster speed, when cardiac locomotor coupling was stronger.

Analogies and differences between cirrhotic cardiomyopathy and hepatopulmonary syndrome.

Cirrhotic cardiomyopathy and hepatopulmonary syndrome are two quite frequent clinical entities that may complicate the course of liver cirrhosis. The common pathophysiological origin and the same clinical presentation make them difficult to compare. Cirrhotic cardiomyopathy and hepatopulmonary syndrome may start with dyspnea and breathlessness but the former is characterized by a chronic cardiac dysfunction and the latter by a defect of oxygenation due to pulmonary shunts formation. The focus is to differentiate them as soon as possible since the treatment is different until the patient undergoes liver transplant that is the real unique cure for them.

Reliability and Agreement of Point and 2-D Shear-Wave Elastography in Assessing the Sciatic Nerve Stiffness.

Information is lacking about the reliability and agreement of different shear-wave elastographic modes in the peripheral nervous system evaluation. The aim of this observational study was to evaluate reproducibility and agreement of two different shear-wave elastographic modes for measuring the sciatic nerve stiffness in patients affected by osteoarthrosis. Two sets of three measurements were conducted bilaterally on the sciatic nerve of 20 patients with point and 2-D shear-wave elastography by a unique expert sonographer. This consecutive case series study was performed in 1 mo. No significant difference was found comparing the first with the second set of evaluations (p = 0.08 for point shear-wave elastography and 0.3 for 2-D shear-wave elastography). Correlation between the two sets of measurements was good and excellent (0.799 with point shear-wave elastography and 0.877 with 2-D shear-wave elastography). Intra-class coefficient correlation between the two sets of measurements was excellent for both shear-wave elastographic modes (0.869 and 0.938, respectively); no agreement between them was demonstrated (analysis of variance [ANOVA] test: p = 0.014).Despite the lack of agreement owing to the different procedures for measuring, both shear-wave elastographic modes allow reliable stiffness measurements of the sciatic nerve and may be used to evaluate stiffness changes.

Accuracy and performance of a new handheld ultrasound machine with wireless system.

We verified the accuracy and performance of a new handheld ultrasound machine, in comparison to a high-end sonographic machine. We performed bilateral measurements of the following tendon districts (supraspinatus, flexor of the middle finger, patellar and Achilles) and of the cross sectional area of the median nerve in 21 patients using a musculoskeletal ultrasound linear scanner of a handheld sonographic machine and a high-end sonographic machine. Two tail T test was used to evaluate whether there were differences in the measurements between the two sonographic machines. Agreement was evaluated by Pearson's correlation. The mean time requested for the examinations was 18 and 9 minutes for the handheld and high-end sonographic machines, respectively. No significant differences were found between the measurements obtained with the handheld ultrasound machine and those with the high-end sonographic machine (p value ranging between 0.31 and 0.97, according to the examined district), whereas, a moderate correlation was found (r coefficient ranging between 0.43 and 0.77, according to the examined district). Although the examination with the handheld ultrasound machine took more time, it showed adequate accuracy and performance; this palmar tool might be also useful in operating rooms.

Strain Ultrasound Elastography in the Achilles Tendon of Ankylosing Spondylitis Patients Treated With Anti-TNF-α: A Preliminary Study.

BACKGROUND/AIM: To compare patients affected by ankylosing spondylitis (AS) treated with anti-TNF-α for two years with controls in terms of Achilles tendon stiffness, ultrasound structure and thickness. PATIENTS AND METHODS: B-mode ultrasound evaluation and strain ultrasound elastography were performed in longitudinal and transverse planes on 22 Achilles tendons of 11 AS patients and 26 of 13 controls. RESULTS: There were no significant differences in thickness and stiffness of the Achilles tendon between AS patients and controls, except for an increased thickness in the middle third of the tendon in the AS patients (p=0.04). The Achilles tendon stiffness ratio of AS patients was 1.02±0.36 vs. 1.14±0.38 in the controls (p=0.2). CONCLUSION: AS patients had an Achilles tendon thickness greater than controls at the middle third, but no difference in the stiffness was found among them. Strain ultrasound elastography may be useful to exclude early changes in mechanical properties of tendons.

Inhibition of Chemoresistance in Primary Tumor Cells by Camellia sinensis non fermentatum Extract Noviphenone (NPE®).

BACKGROUND/AIM: Despite improvements in cancer therapy, life expectancy after tumor recurrence remains low. Relapsed cancer is characterized by drug resistance, often mediated through overexpression of multidrug resistance (MDR) genes. Camellia sinensis non fermentatum extract is known for its anticancer properties in several cancer cell lines and might improve cancer therapy outcome after tumor recurrence. MATERIALS AND METHODS: Embryonal rhabdomyosarcoma cell lines, alveolar rhabdomyosarcoma cell lines and primary rhabdomyosarcoma MAST139 cells were used to test NPE® effects on cell viability in combination with chemotherapeutic agents. Cell viability was measured by the WST-1 assay and CV staining. Gene expression levels of chemotherapy-induced efflux pumps and their activity was assessed upon NPE® treatment by measuring doxorubicin retention through evaluation of the autofluorescence signal. RESULTS: Administration of increasing doxorubicin concentrations triggered immediate adaptation to the drug, which was surprisingly overcome by the addition of NPE®. Investigating the mechanism of immediate adaptation, MDR1 gene overexpression was observed upon doxorubicin treatment. Although NPE® did not alter pump gene expression, it was able to reduce pump activity, thus allowing the chemotherapeutic agent to stay inside the cells to exert its full anticancer activity. CONCLUSION: NPE® might improve chemotherapeutic treatment by re-sensitizing relapsed tumors to anticancer drugs. Fighting MDR represents the key to overcome tumor relapse and improve the overall survival of cancer patients.

Reliability of quantitative point shear-wave ultrasound elastography on vastus medialis muscle and quadriceps and patellar tendons.

AIM: The aim of this work was to evaluate the reliability of pSWE in assessing the stiffness of the vastus medialis muscle and of the quadriceps and patellar tendons. MATERIAL AND METHODS: For this purpose, 18 subjects (9 males and 9 females of 57±22 years) in good clinical conditions were included in this study. pSWE examination was conducted by a unique expert operator with more than ten years of experience in musculoskeletal ultrasound. Two sets of five measurements for each muscle and tendon district were bilaterally performed at the same manner, at least fifteen minutes apart. The mean value of the measurements of each set was statistically compared with that of the other set. RESULTS: No significant differences were found comparing the mean value of the measurements of the two sets of evaluation performed in muscle and tendon areas (vastus medialis muscle: p=0.285; quadriceps tendon: p=0.979; patellar tendon: p=0.187). The intraclass correlation coefficient was excellent for all areas (vastus medialis muscle: 0.969; quadriceps and patellar tendons: 0.995 and 0.989, respectively). CONCLUSION:  The pSWE technique demonstrated that it was a reliable method for measuring stiffness in vastus medialis muscle and quadriceps and patellar tendon in subjects who had undergone orthopedic surgery. This opens the possibility of many applications in monitoring stiffness before and after surgery and during rehabilitation.

Differences in carotid atherosclerosis between patients with ankylosing spondylitis treated with tumor necrosis factor-α antagonists and healthy matched controls.

An increased vascular risk is present in patients with ankylosing spondylitis (AS). In this report, we evaluate the presence and grade of atherosclerosis in patients with AS, uninterruptedly treated with tumor necrosis factor-α (TNF-α) antagonists for 2 years, in comparison to that in a nontreated group of healthy controls.Fourteen patients with AS and 14 healthy controls underwent carotid sonography to measure intima-media thickness (IMT) and to evaluate the presence of plaque. Bath Ankylosing Spondylitis Disease Activity Index, Bath Ankylosing Spondylitis Metrology Index, Bath Ankylosing Spondylitis Functional Index scores, erythrocyte sedimentation rate, C-reactive protein, glycemia, total cholesterol, and triglyceride levels were also recorded.Patients with AS showed significantly lower values of mean and maximum IMT at the level of the common carotid (P = .02 and .04, respectively) and the carotid bulb (P = .0006 and .0005, respectively) compared to those of healthy controls. They also had a number of carotid plaques significantly lower than that of healthy controls (P = .02). No differences were found in IMT values at the level of internal carotid between the 2 populations.The significantly lower carotid atherosclerosis found in patients with AS treated with TNF antagonists than in healthy controls shows the important complementary role of this treatment in reducing vascular disease progression probably by decreasing inflammation.

Diagnostic approach to neuropsychiatric lupus erythematosus: what should we do?

Neuropsychiatric systemic lupus erythematosus is a diagnostic challenge due to the multifarious neurological and psychiatric manifestations that define it but, when suspected, diagnostic imaging can give a fundamental help. The advancements and variety of neuroimaging techniques allow us to perform more and more accurate evaluations of structure, perfusion, and metabolism of the brain and to detect cerebral and spinal lesions. Moreover, vascular districts of the neck and the brain, as well as the electrical brain and peripheral muscle activity may be accurately investigated, thus giving us a wide panoramic view. Although magnetic resonance is recognized as a fundamental neuroimaging technique to reach a correct diagnosis, the juxtaposition of other diagnostic techniques has improved the possibility to make diagnoses but has also increased the confusion about deciding which of them to use and when. Our aim was to combine the number of available techniques with the need to simplify the diagnostic path. Therefore, through the construction of an algorithm from an evidence based approach, we believe we are providing some added improvements to facilitate and expedite the diagnosis of NPSLE.

An ultrasound automated method for non-invasive assessment of carotid artery pulse wave velocity.

To validate the clinical applicability and feasibility of an automated ultrasound (US) method in measuring the arterial stiffness of patients with chronic inflammatory rheumatic diseases, comparing automated measurements performed by a rheumatologist without experience in vascular sonography with those obtained by a sonographer experienced in vascular US, using a standardized manual method. Twenty subjects affected by different chronic inflammatory rheumatic disorders were consecutively recruited. For each patient, the arterial stiffness of both common carotids was manually calculated. Subsequently, the measure of the pulse wave velocity (PWV) was obtained using an US device called Radio Frequency - Quality Arterial Stiffness (RF-QAS), provided by the same US system (ie, My Lab 70 XVG, Esaote SpA, Genoa, Italy) equipped with a 4-13 MHz linear probe. The reliability comparison between the two US methods was calculated using the intraclass correlation coefficient (ICC). ICC between the values obtained with the two methods for calculating the arterial stiffness resulted 0.789. A significant positive correlation between the two methods was also established with Pearson's (r=0.62, p<0.0001) and Spearman's analysis (r=0.66, p=0.001). A significant performance comparison was seen using Bland-Altman plot. The acquisition of the arterial stiffness parameter with the automated method required about 2 min for each patient. Clinical applicability of this US automated method to assess PWV at common carotid level by a rheumatologist is reliable and feasible in comparison with a conventional manual method.

Portopulmonary hypertension and hepatorenal syndrome. Two faces of the same coin.

Portopulmonary hypertension and hepatorenal syndrome are both severe local hypertensive complications of liver cirrhosis and portal hypertension. Both are characterized by vasoconstrictive manifestations regarding pulmonary and renal vascular network, respectively. This review addresses the mechanisms underlying the development of vasoconstriction that leads to local vascular hypertension in the lung and in the kidney with the result of organ dysfunction. Potential therapeutic options are available for the management of these two syndromes as a bridge for liver transplantation; clinical efficacy depends in part on the time and rapidity of intervention and in part on how serious the chain of events is that has triggered the entire vasoconstrictive process.

Targeting the EWS-ETS transcriptional program by BET bromodomain inhibition in Ewing sarcoma.

Ewing sarcomas (ES) are highly malignant bone or soft tissue tumors. Genetically, ES are defined by balanced chromosomal EWS/ETS translocations, which give rise to chimeric proteins (EWS-ETS) that generate an oncogenic transcriptional program associated with altered epigenetic marks throughout the genome. By use of an inhibitor (JQ1) blocking BET bromodomain binding proteins (BRDs) we strikingly observed a strong down-regulation of the predominant EWS-ETS protein EWS-FLI1 in a dose dependent manner. This was further enhanced by co-treatment with an inhibitor of the PI3K pathway. Microarray analysis further revealed JQ1 treatment to block a typical ES associated expression program. The effect on this expression program was mimicked by RNA interference with BRD3 or BRD4 expression, indicating that the EWS-FLI1 mediated expression profile is at least in part mediated via such epigenetic readers. Consequently, contact dependent and independent proliferation of different ES lines was strongly inhibited. Mechanistically, treatment of ES resulted in a partial arrest of the cell cycle as well as induction of apoptosis. Tumor development was suppressed dose dependently in a xeno-transplant model in immune deficient mice, overall indicating that ES may be susceptible to treatment with epigenetic inhibitors blocking BET bromodomain activity and the associated pathognomonic EWS-ETS transcriptional program.

PI3K/AKT signaling modulates transcriptional expression of EWS/FLI1 through specificity protein 1.

Ewing sarcoma (ES) is the second most frequent bone cancer in childhood and is characterized by the presence of the balanced translocation t(11;22)(q24;q12) in more than 85% of cases, generating a dysregulated transcription factor EWS/FLI1. This fusion protein is an essential oncogenic component of ES development which is necessary for tumor cell maintenance and represents an attractive therapeutic target. To search for modulators of EWS/FLI1 activity we screened a library of 153 targeted compounds and identified inhibitors of the PI3K pathway to directly modulate EWS/FLI1 transcription. Surprisingly, treatment of four different ES cell lines with BEZ235 resulted in down regulation of EWS/FLI1 mRNA and protein by ~50% with subsequent modulation of target gene expression. Analysis of the EWS/FLI1 promoter region (-2239/+67) using various deletion constructs identified two 14 bp minimal elements as being important for EWS/FLI1 transcription. We identified SP1 as modulator of EWS/FLI1 gene expression and demonstrated direct binding to one of these regions in the EWS/FLI1 promoter by EMSA and ChIP experiments. These results provide the first insights on the transcriptional regulation of EWS/FLI1, an area that has not been investigated so far, and offer an additional molecular explanation for the known sensitivity of ES cell lines to PI3K inhibition.