Quantitative Evaluation of Stance as a Sensitive Biomarker of Postural Ataxia Development in Preclinical SCA1 Mutation Carriers.

The aim of this study was to determine the time between the first detection of postural control impairments and the evident manifestation of ataxia in preclinical SCA1 individuals. Twenty five preclinical SCA1 mutation carriers: 13 with estimated disease onset ≤ 6 years (SCA1 +) aged 27.8 ± 8.1 years; 12 with expected disease onset > 6 years (SCA1-) aged 26.6 ± 3.1 years and 26 age and sex matched healthy controls (HCs) underwent static posturography during 5 years of observation. The movements of the centre of feet pressure (COP) during quiet standing with eyes open (EO) and closed (EC) were quantified by calculating the mean radius (R), developed surface area (A) and mean COP movement velocity (V). Ataxia was evaluated by use of the Scale for Assessment and Rating of Ataxia (SARA).SCA1 + exhibited significantly worse quality of stance with EC vs. SCA1- (p < 0.05 for V) and HCs (p < 0.001) even 5 to 6 years before estimated disease onset. There were no statistically significant differences between SCA1- and HCs. A slow increase in Cohen's d effect size was observed for VEO up to the clinical manifestation of ataxia. VEO and AEC recorded in preclinical SCA1 individuals correlated slightly but statistically significantly with SARA (r = 0.47).The study confirms that static posturography detects COP sway changes in SCA1 preclinical gene carriers even 5 to 6 years before estimated disease onset. The quantitative evaluation of stance in preclinical SCA is a sensitive biomarker for the monitoring of the disease progression and may be useful in clinical trials.

Quantitative Oculomotor Assessment in Hereditary Ataxia: Discriminatory Power, Correlation with Severity Measures, and Recommended Parameters for Specific Genotypes.

Characterizing bedside oculomotor deficits is a critical factor in defining the clinical presentation of hereditary ataxias. Quantitative assessments are increasingly available and have significant advantages, including comparability over time, reduced examiner dependency, and sensitivity to subtle changes. To delineate the potential of quantitative oculomotor assessments as digital-motor outcome measures for clinical trials in ataxia, we searched MEDLINE for articles reporting on quantitative eye movement recordings in genetically confirmed or suspected hereditary ataxias, asking which paradigms are most promising for capturing disease progression and treatment response. Eighty-nine manuscripts identified reported on 1541 patients, including spinocerebellar ataxias (SCA2, n = 421), SCA3 (n = 268), SCA6 (n = 117), other SCAs (n = 97), Friedreich ataxia (FRDA, n = 178), Niemann-Pick disease type C (NPC, n = 57), and ataxia-telangiectasia (n = 85) as largest cohorts. Whereas most studies reported discriminatory power of oculomotor assessments in diagnostics, few explored their value for monitoring genotype-specific disease progression (n = 2; SCA2) or treatment response (n = 8; SCA2, FRDA, NPC, ataxia-telangiectasia, episodic-ataxia 4). Oculomotor parameters correlated with disease severity measures including clinical scores (n = 18 studies (SARA: n = 9)), chronological measures (e.g., age, disease duration, time-to-symptom onset; n = 17), genetic stratification (n = 9), and imaging measures of atrophy (n = 5). Recurrent correlations across many ataxias (SCA2/3/17, FRDA, NPC) suggest saccadic eye movements as potentially generic quantitative oculomotor outcome. Recommendation of other paradigms was limited by the scarcity of cross-validating correlations, except saccadic intrusions (FRDA), pursuit eye movements (SCA17), and quantitative head-impulse testing (SCA3/6). This work aids in understanding the current knowledge of quantitative oculomotor parameters in hereditary ataxias, and identifies gaps for validation as potential trial outcome measures in specific ataxia genotypes.

Quantitative Speech Assessment in Ataxia-Consensus Recommendations by the Ataxia Global Initiative Working Group on Digital-Motor Markers.

Dysarthria is a common and debilitating symptom of many neurodegenerative diseases, including those resulting in ataxia. Changes to speech lead to significant reductions in quality of life, impacting the speaker in most daily activities. Recognition of its importance as an objective outcome measure in clinical trials for ataxia is growing. Its viability as an endpoint across the disease spectrum (i.e. pre-symptomatic onwards) means that trials can recruit ambulant individuals and later-stage individuals who are often excluded because of difficulty completing lower limb tasks. Here we discuss the key considerations for speech testing in clinical trials including hardware selection, suitability of tasks and their role in protocols for trials and propose a core set of tasks for speech testing in clinical trials. Test batteries could include forms suitable for remote short, sensitive and easy to use, with norms available in several languages. The use of artificial intelligence also could improve accuracy and automaticity of analytical pipelines in clinic and trials.

Consistency of the results of neurophysiological examinations with clinical diagnosis formed by the referring physician in ambulatory medical care.

Purpose: Access to electroneurographic/electromyographic (ENG/EMG) examinations and the number of patients referred for electrodiagnostic (EDX) examination are increasing. We aimed to determine the accuracy of the initial clinical diagnosis made by outpatient medical care physicians who referred patients to the EMG laboratory. Methods: We analyzed referrals and EDX results of all patients who visited EMG laboratory of the Department of Clinical Neurophysiology, Institute of Psychiatry and Neurology in Warsaw in 2021. Examinations were performed in accordance with the standards and norms adopted in our laboratory by EMG-certified neurologists, regarding the initial diagnosis stated by referring physicians. Results: A total of 454 EDX results from 412 patients were analyzed. Most of patients (54.6%) were referred with diagnosis of carpal tunnel syndrome (CTS), followed by single nerves damage (18.7%), polyneuropathy (18.1%), tetany (7.0%), myasthenia gravis (1.3%) or myopathy (0.2%). The result of the ENG/EMG examination was: diagnosis confirmation (61.9%), a new clinically significant diagnosis or additional asymptomatic nerve damage (32.4%), and normal examination result (25.1%) of patients. Electrophysiological examination most often confirmed the referral diagnosis in patients with suspected CTS (75.4%), followed by single nerves damage (51.8%), polyneuropathy (48.8%), tetany (31.3%) and the least for myasthenia gravis and myopathy (0%). Conclusions: Our study showed frequent inconsistency of the EDX results with the clinical diagnosis formed by the referring physician. A high percentage of normal test results was noted. Initial diagnosis and the scope of EDX examination should be determined by detailed interview and physical examination.

Quantitative Oculomotor Assessment in Hereditary Ataxia: Systematic Review and Consensus by the Ataxia Global Initiative Working Group on Digital-motor Biomarkers.

Oculomotor deficits are common in hereditary ataxia, but disproportionally neglected in clinical ataxia scales and as outcome measures for interventional trials. Quantitative assessment of oculomotor function has become increasingly available and thus applicable in multicenter trials and offers the opportunity to capture severity and progression of oculomotor impairment in a sensitive and reliable manner. In this consensus paper of the Ataxia Global Initiative Working Group On Digital Oculomotor Biomarkers, based on a systematic literature review, we propose harmonized methodology and measurement parameters for the quantitative assessment of oculomotor function in natural-history studies and clinical trials in hereditary ataxia. MEDLINE was searched for articles reporting on oculomotor/vestibular properties in ataxia patients and a study-tailored quality-assessment was performed. One-hundred-and-seventeen articles reporting on subjects with genetically confirmed (n=1134) or suspected hereditary ataxia (n=198), and degenerative ataxias with sporadic presentation (n=480) were included and subject to data extraction. Based on robust discrimination from controls, correlation with disease-severity, sensitivity to change, and feasibility in international multicenter settings as prerequisite for clinical trials, we prioritize a core-set of five eye-movement types: (i) pursuit eye movements, (ii) saccadic eye movements, (iii) fixation, (iv) eccentric gaze holding, and (v) rotational vestibulo-ocular reflex. We provide detailed guidelines for their acquisition, and recommendations on the quantitative parameters to extract. Limitations include low study quality, heterogeneity in patient populations, and lack of longitudinal studies. Standardization of quantitative oculomotor assessments will facilitate their implementation, interpretation, and validation in clinical trials, and ultimately advance our understanding of the evolution of oculomotor network dysfunction in hereditary ataxias.

In silico assessment of risks associated with pesticides exposure during pregnancy.

Novel Quantitative Structure-Activity Relationship (QSAR) models of compounds' placenta (PL) permeability expressed as their log FM (fetus-to-mother blood concentration) values or binary PL1/0 (crossing/non-crossing) score were generated using a number of statistical tools: Multiple Linear Regression, Boosted Trees, Principal Component Analysis and Artificial Neural Networks, on the basis of molecular descriptors calculated by Mordred software and selected using Partial Least Squares (PLS) analysis. It was established that the most important predictor of both log FM and the binary PL1/0 score is Lipinski - a binary variable reflecting the compounds' ability to satisfy the criteria of drug-likeness according to the Lipinski's "Rule of 5". The quantitative (log FM) and qualitative (PL1/0) models of PL permeability were applied to 345 pesticides from different chemical families (triazines, carbamates, pyrethroids, organochlorine, organophosphorus and miscellaneous compounds). The ability of studied pesticides to cross the placenta was assessed; the basic physico-chemical parameters responsible for good or poor placenta transport of pesticides were identified and the relationships between the pesticides' PL permeability, blood-brain barrier (BBB) transfer and gastro-intestinal (GI) absorption were investigated. It was found (on the basis of logistic regression analysis) that the probability of a compound crossing the placenta (PL1) is inversely correlated with its lipophilicity and molar refractivity and positively correlated with the total count of oxygen and nitrogen atoms.

Immobilized Keratin HPLC Stationary Phase-A Forgotten Model of Transdermal Absorption: To What Molecular and Biological Properties Is It Relevant?

Chromatographic retention data collected on immobilized keratin (KER) or immobilized artificial membrane (IAM) stationary phases were used to predict skin permeability coefficient (log Kp) and bioconcentration factor (log BCF) of structurally unrelated compounds. Models of both properties contained, apart from chromatographic descriptors, calculated physico-chemical parameters. The log Kp model, containing keratin-based retention factor, has slightly better statistical parameters and is in a better agreement with experimental log Kp data than the model derived from IAM chromatography; both models are applicable primarily to non-ionized compounds.Based on the multiple linear regression (MLR) analyses conducted in this study, it was concluded that immobilized keratin chromatographic support is a moderately useful tool for skin permeability assessment.However, chromatography on immobilized keratin may also be of use for a different purpose-in studies of compounds' bioconcentration in aquatic organisms.

Immobilized artificial membrane-chromatographic and computational descriptors in studies of soil-water partition of environmentally relevant compounds.

Chromatographic retention factor log kIAM obtained from immobilized artificial membrane (IAM) HPLC with buffered, aqueous mobile phases and calculated molecular descriptors (molecular weight - log MW; molar volume - VM; polar surface area - PSA; total count of nitrogen and oxygen atoms -(N + O); count of freely rotable bonds - FRB; H-bond donor count - HD; H-bond acceptor count - HA; energy of the highest occupied molecular orbital - EHOMO; energy of the lowest unoccupied orbital - ELUMO; dipole moment - DM; polarizability - α) obtained for a group of 175 structurally unrelated compounds were tested in order to generate useful models of solutes' soil-water partition coefficient normalized to organic carbon log Koc. It was established that log kIAM obtained in the conditions described in this study is not sufficient as a sole predictor of the soil-water partition coefficient. Simple, potentially useful models based on log kIAM and a selection of readily available, calculated descriptors and accounting for over 88% of total variability were generated using multiple linear regression (MLR) and artificial neural networks (ANN). The models proposed in the study were tested on a group of 50 compounds with known experimental log Koc values by plotting the calculated vs. experimental values. There is a good close similarity between the calculated and experimental data for both MLR and ANN models for compounds from different chemical families (R2 ≥ 0.80, n = 50) which proves the models' reliability.

Affinity of Compounds for Phosphatydylcholine-Based Immobilized Artificial Membrane-A Measure of Their Bioconcentration in Aquatic Organisms.

The BCF (bioconcentration factor) of solutes in aquatic organisms is an important parameter because many undesired chemicals enter the ecosystem and affect the wildlife. Chromatographic retention factor log kwIAM obtained from immobilized artificial membrane (IAM) HPLC chromatography with buffered, aqueous mobile phases and calculated molecular descriptors obtained for a group of 120 structurally unrelated compounds were used to generate useful models of log BCF. It was established that log kwIAM obtained in the conditions described in this study is not sufficient as a sole predictor of bioconcentration. Simple, potentially useful models based on log kwIAM and a selection of readily available, calculated descriptors and accounting for over 88% of total variability were generated using multiple linear regression (MLR), partial least squares (PLS) regression and artificial neural networks (ANN). The models proposed in the study were tested on an external group of 120 compounds and on a group of 40 compounds with known experimental log BCF values. It was established that a relatively simple MLR model containing four independent variables leads to satisfying BCF predictions and is more intuitive than PLS or ANN models.

C9orf72 hexanucleotide repeat expansion found in suspected spinobulbar muscular atrophy (SBMA).

INTRODUCTION: The expansion of a hexanucleotide GGGGCC repeat (G4C2) in the C9orf72 locus is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). In addition, C9orf72 expansion has also been detected in patients with a clinical manifestation of Parkinson's Disease (PD), Alzheimer's Disease (AD), Huntington's Disease (HD), and ataxic disorders. MATERIAL AND METHODS: A total of 1,387 patients with clinically suspected ALS, HD or spinal and bulbar muscular atrophy (SBMA) were enrolled, and the prevalence of C9orf72 expansions was estimated. RESULTS: The hexanucleotide expansion accounted for 3.7% of the ALS patients, 0.2% of the HD suspected patients with excluded HTT mutation, and 1.3% of the suspected SBMA patients with excluded mutation in AR gene. CONCLUSIONS: This is the first report revealing the presence of C9orf72 expansion in patients with a suspected SBMA diagnosis. Consequently, we advise testing for C9orf72 expansion in patients presenting with the SBMA phenotype and a genetically unsolved diagnosis.

IAM Chromatographic Models of Skin Permeation.

Chromatographic retention factor log kIAM obtained from IAM HPLC chromatography with buffered aqueous mobile phases and calculated molecular descriptors (surface area-Sa; molar volume-VM; polar surface area-PSA; count of freely rotable bonds-FRB; H-bond acceptor count-HA; energy of the highest occupied molecular orbital-EHOMO; energy of the lowest unoccupied orbital-ELUMO; and polarizability-α) obtained for a group of 160 structurally unrelated compounds were tested in order to generate useful models of solutes' skin permeability coefficient log Kp. It was established that log kIAM obtained in the conditions described in this study is not sufficient as a sole predictor of the skin permeability coefficient. Simple put, potentially useful models based on log kIAM and readily available calculated descriptors, accounting for 85 to 91% of the total variability, were generated using Multiple Linear Regression (MLR).The models proposed in the study were tested on a group of 20 compounds with known experimental log Kp values.

RP-18 TLC and Computational Descriptors of Skin Permeability of Sunscreens.

The relationships between the reversed-phase thin layer chromatographic retention parameters obtained on octadecyl-modified silica (RP-18) sorbent for mobile phases containing water and one of six water-miscible organic modifiers (acetone, methanol, acetonitrile, tetrahydrofurane, N,N-dimethylformamide, 1,4-dioxane) and skin permeability coefficients were studied for a group of 21 cosmetic raw materials, mainly organic sunscreens and preservatives. The correlations between the skin permeability coefficients log Kd calculated in silico using EpiSuite software and the RP-18 thin layer chromatographic retention parameters are mostly linear, especially for compounds of lower-to-medium lipophilicity. It was established that skin permeability coefficient models based on retention parameters collected for mobile phases containing acetone or dioxane (75% v/v), proposed for structurally unrelated cosmetic raw materials are also applicable to other actives, as shown using a test set of compounds whose in vivo log Kd data are available. Skin permeability models developed in this study have the benefit of being based on easily obtained, chromatographic descriptors and their applicability extends beyond cosmetic chemistry.

RP-18 TLC Chromatographic and Computational Study of Skin Permeability of Steroids.

The skin permeability of steroids, as investigated in this study, is important because some of these compounds are, or could, be used in preparations applied topically. Several models of skin permeability, involving thin layer chromatographic and calculated descriptors, were generated and validated using Kp reference values obtained in silico and then tested on a group of solutes whose experimental Kp values could be found (log Kpexp). The study established that the most applicable log Kp model is based on RP-18 thin layer chromatographic data (RM) and the calculated descriptors VM (molar volume) and PSA (polar surface area). Two less efficient, yet simple, equations based on PSA or VM combined with HD (H-donor count) can be used with caution for rapid, rough estimations of compounds' skin permeability prior to their chemical synthesis.

Evaluation of drug-likeness and ADME properties of sunscreens and preservatives using reversed-phase thin layer chromatographic retention data and calculated descriptors.

RP-18 TLC chromatography was used to evaluate the pharmacokinetic properties (volume of distribution, VD; plasma protein binding, %PPB; the ability to cross the blood-brain barrier expressed as log PS and log BB) of several cosmetic raw materials - sunscreen and preservatives. The majority of these compounds are intended for topical use on skin and their drug-likeness and the ability to cross biological barriers are undesired. The retention parameters RM0, S, PC1 and RM75 % obtained for mobile phases containing six organic modifiers (methanol, acetonitrile, THF, acetone, dioxane, DMF) were used as the sole descriptors or combined with calculated physicochemical properties (PSA, MW, VM) of studied compounds. The chromatographic parameters considered in this study are, generally speaking, good predictors of the compounds' pharmacokinetic properties VD, %PPB and log PS. RM75 % and the novel parameters derived from it (RM75 %/MW and RM75 %/VM) can be considered time- and cost-effective alternatives to the chromatographic parameters obtained by extrapolation or interpolation methods. In the case of some pharmacokinetic properties investigated in this study additional descriptors (PSA) have a significant influence on the quality of correlations.

RP-18 TLC retention data and calculated physico-chemical parameters as predictors of soil-water partition and bioconcentration of organic sunscreens.

RP-18 TLC chromatography was used to evaluate the impact on the environment (mobility in soil expressed as soil-water partition coefficient, log Koc; bioconcentration factor in aquatic organisms, log BCF) of several cosmetic raw materials - sunscreens, preservatives and vitamins. The retention parameters RM0 (RM extrapolated to zero concentration of an organic modifier in a mobile phase), S (slope), PC1 (1st principal component) and RM75% (single TLC run parameter for mobile phases containing 75% (v/v) of an organic modifier) obtained for six organic modifiers (methanol, acetonitrile, THF, acetone, dioxane, DMF) were used as the sole descriptors or combined with calculated physico-chemical properties (PSA - polar surface area; MW - molecular weight; VM - molar volume) of studied compounds. The chromatographic parameters considered in this study are, generally speaking, good predictors of the compounds' mobility in soil or the affinity for aquatic organisms. The parameters RM0, S and RM75% obtained for THF, dioxane and acetone may be used to investigate even very lipophilic compounds. RM75% is of a little bit limited use but it should be considered a time- and cost-effective alternative to the chromatographic parameters obtained by extrapolation or interpolation methods. In the case of some environmental parameters investigated in this study additional descriptors (PSA) have a significant influence on the quality of correlations.

Application of RP-18 TLC Retention Data to the Prediction of the Transdermal Absorption of Drugs.

Several chromatographic parameters (RM0 and S obtained from RP-18 TLC with methanol-pH 7.4 phosphate buffer mobile phases by extrapolation to zero concentration of methanol; Rf and RM obtained from RP-18 TLC with acetonitrile-pH 7.4 phosphate buffer 70:30 v/v as a mobile phase) and calculated molecular descriptors (molecular weight-MW; molar volume-VM; polar surface area-PSA; total count of nitrogen and oxygen atoms-(N+O); H-bond donor count-HD; H-bond acceptor count-HA; distribution coefficient-log D; total energy-ET; binding energy-Eb; hydration energy-Eh; energy of the highest occupied molecular orbital-EHOMO; energy of the lowest unoccupied orbital-ELUMO; electronic energy-Ee; surface area-Sa; octanol-water partition coefficient-log P; dipole moment-DM; refractivity-R, polarizability-α) and their combinations (Rf/PSA, RM/MW, RM/VM) were tested in order to generate useful models of solutes' skin permeability coefficient log Kp. It was established that neither RM0 nor S obtained in the conditions used in this study is a good predictor of the skin permeability coefficient. The chromatographic parameters Rf and Rf/PSA were also unsuitable for this purpose. A simple and potentially useful, purely computational model based on (N+O), log D and HD as independent variables and accounting for ca. 83% of total variability was obtained. The evaluation of parameters derived from RM (RM, RM/MW, RM/VM) as independent variables in log Kp models proved that RM/VM is the most suitable descriptor belonging to this group. In a search for a reliable log Kp model based on this descriptor two possibilities were considered: a relatively simple model based on 5 independent variables: (N+O), log D, RM/VM, ET and Eh and a more complex one, involving also Eb, MW and PSA.

Clinical correlates of rhinovirus infection in preschool asthma.

BACKGROUND: Investigation of preschool asthma is important since not all children outgrow their illness during this age. Data are scarce on the role of rhinovirus (RV) infections in this patient group. OBJECTIVES: To investigate the role of RV infections in preschool asthma: (i) susceptibility factors, (ii) clinical course, and (iii) medium-term outcome. METHODS: A total of 130 asthmatic children aged 4-6 years from the multinational PreDicta cohort were prospectively followed for a 12-month period. Allergy tests and a standard health questionnaire were carried out at study entry. Respiratory virus presence in nasopharyngeal washes was studied at illness visits and at 3 scheduled visits. RESULTS: At study entry, mean age of the children was 5.3 years. Of 571 visits, 54% were positive for any virus and 39% for RV. Patient characteristics were only assessed with RV infection due to low number of other viruses. The use of supplementary vitamin D was inversely associated with RV infection (P < .05). RV infection was associated with more severe course of acute illness in terms of more severe nighttime coughing, more sleep disturbances, and more days with runny nose (all P < .05). RV infection was also associated with more severe disease course during the 12-month follow-up in terms of more nights with awakenings and more days of exercise-related symptoms (both P < .05). CONCLUSIONS: Vitamin D supplementation may have an anti-rhinovirus effect. Both short- and medium-term outcomes suggest RV infection to be an important clinical marker of instable preschool asthma.

Emerging or Underestimated Silica-Based Stationary Phases in Liquid Chromatography.

Several newly synthesized or forgotten silica-based stationary phases proposed for liquid chromatography are described, including non-endcapped, short-chain alkyl phases; hydrophilic and polar-endcapped stationary phases; polar-embedded alkyl phases; long-chain alkyl phases. Stationary phases with aromatic, cyanopropyl, diol and aminopropyl functionalities are also reviewed. Stationary phases of particular interest are biomolecular materials - based on immobilized cholesterol, aminoacids, peptides, proteins or lipoproteins. Packing materials involving macrocyclic chemistry (crown ethers; calixarenes; aza-macrocycles; oligo-and polysaccharides including these of marine origin - chitin- or chitosan-based; macrocyclic antibiotics) are discussed. Since many stationary phases developed for one type of applications (e.g. chiral separation) have been found useful in solving other analytical problems (e.g. drug's plasma protein binding ability), it seemed reasonable to discuss particular chemistries behind the stationary phases presented in this review rather than specific types of interactions or chromatographic modes.

Next-generation sequencing study reveals the broader variant spectrum of hereditary spastic paraplegia and related phenotypes.

Hereditary spastic paraplegias (HSPs) are clinically and genetically heterogeneous neurodegenerative disorders. Numerous genes linked to HSPs, overlapping phenotypes between HSP subtypes and other neurodegenerative disorders and the HSPs' dual mode of inheritance (both dominant and recessive) make the genetic diagnosis of HSPs complex and difficult. Out of the original HSP cohort comprising 306 index cases (familial and isolated) who had been tested according to "traditional workflow/guidelines" by Multiplex Ligation-dependent Probe Amplification (MLPA) and Sanger sequencing, 30 unrelated patients (all familial cases) with unsolved genetic diagnoses were tested using next-generation sequencing (NGS). One hundred thirty-two genes associated with spastic paraplegias, hereditary ataxias and related movement disorders were analysed using the Illumina TruSight™ One Sequencing Panel. The targeted NGS data showed pathogenic variants, likely pathogenic variants and those of uncertain significance (VUS) in the following genes: SPAST (spastin, SPG4), ATL1 (atlastin 1, SPG3), WASHC5 (SPG8), KIF5A (SPG10), KIF1A (SPG30), SPG11 (spatacsin), CYP27A1, SETX and ITPR1. Out of the nine genes mentioned above, three have not been directly associated with the HSP phenotype to date. Considering the phenotypic overlap and joint cellular pathways of the HSP, spinocerebellar ataxia (SCA) and amyotrophic lateral sclerosis (ALS) genes, our findings provide further evidence that common genetic testing may improve the diagnostics of movement disorders with a spectrum of ataxia-spasticity signs.

Aspects of Drug-Protein Binding and Methods of Analyzing the Phenomenon.

In recent decades, drug-protein interactions have been widely studied and several methods of analysis of these phenomena have been developed and improved. These can be classified into separation, physical, chromatographic and electrophoretic methods. This review depicts the assumptions and mechanisms of methods from each group, details their strengths and weaknesses, and presents examples of their usage from the literature. Equilibrium dialysis, ultrafiltration, Hummel-Dreyer method or high performance affinity chromatography are given as representative examples, but this issue is far more expanded. Nowadays, increasing attention is paid to the computational methods and molecular modeling which are convenient tools to estimate protein binding affinity based on the physicochemical properties of compounds. To gain a broader overview, the study also examines the protein binding ability and pharmacotherapy of drugs against a range of substrates such as plasma, skin, tissue and human milk.