Hungarian adaptation of the cerebellar cognitive affective/Schmahmann Syndrome Scale.

Recent studies have reported that cerebellar lesions can cause cognitive, behavioral, and affective symptoms. This constellation is called the cerebellar cognitive affective syndrome (CCAS). A bedside instrument, the CCAS-Scale, has been developed to screen for this clinical presentation. The aim of this study is to adapt the CCAS-Scale to Hungarian according to international cross-cultural guidelines. In cooperation with the senior author of the original CCAS-Scale, we defined a five-step adaptation protocol (license number 6758-1/2021). Step 1: translation of the scale from English to Hungarian by two separate teams. Step 2: comparison of the two translated versions, synthesis (preliminary version). Step 3: back translation by an independent professional translator. Step 4: authorization, revision, and correction. Step 5: pre-testing the scale, measuring the test times. Following our protocol, we produced the CCAS-H and the instructions booklet. We pre-tested healthy (n = 10) and cerebellar stroke patients (n = 10) and finalized the scale. Although not significantly, but cerebellar patients reached lower raw scores compared with healthy subjects. Testing times differed significantly between the two groups. A meticulous validation protocol was outlined to assess the validity and reliability of the newly adapted test. CCAS-H is a quick and adequate scale to examine the cerebellar-cognitive affective syndrome, which will be available for Hungarian professionals. Our main challenge was to define the stimuli and cues with adequate psycholinguistic and psychometric properties. As a next step, we are gathering data for the validation with the help of six other Hungarian Neurology departments.

Aspiration Risk Screening With Tongue Pressure Measurement in Acute Stroke: A Diagnostic Accuracy Study Using STARD Guidelines.

Introduction: Dysphagia can affect more than 50% of stroke patients in the acute phase. Aspiration pneumonia is a serious complication that can be prevented with dysphagia screening and assessment. Measurement of tongue elevation pressure is suggested to be a useful tool in aspiration risk screening. Objective: This study aimed to assess the diagnostic accuracy of maximum anterior tongue elevation strength (Pmax) in acute stroke care. Method: In this prospective study, data were collected in a neurology department (stroke center) where patients formed a consecutive case series. The sample consisted of thirty stroke patients who failed an initial dysphagia screening. Patients underwent anterior tongue elevation strength measurement (index test) during bedside dysphagia assessment by a speech-language pathologist and flexible endoscopic evaluation of swallowing (reference test) by an otorhinolaryngologist on the same day. Outcome variables (index values in kPa, reference values interpreted on the penetration-aspiration scale) were used for estimating measures of diagnostic accuracy in aspiration risk screening. Results: Ten patients aspirated on instrumental evaluation. At the cut-off point of ≤ 34 kPa the analysis showed 90% sensitivity, 35% specificity, 41% positive predictive value, and 88% negative predictive value. The area under the curve (AUC) for Pmax was AUC = 0.700 (95% CI [0.500-0.900]). Conclusion: Although individuals with low anterior tongue elevation strength tend to have a higher risk of aspiration, this variable alone is not capable of screening aspiration in acute stroke. In combination with a thorough noninstrumental bedside examination, it might have the potential to reduce the number of false positive cases. Further studies in this area would be worthwhile.

Application of Normal-Phase Silica Column in Hydrophilic Interaction Liquid Chromatography Mode for Simultaneous Determination of Underivatized Amino Acids from Human Serum Samples via Liquid Chromatography-Tandem Mass Spectrometry.

In neonatal screening, amino acids have a significant diagnostic role. Determination of their values may identify abnormal conditions. Early diagnosis and continuous monitoring of amino acid disorders results in a better disease outcome. An easy and simple LC-MS/MS method was developed for the quantitation of underivatized amino acids. Amino acids were separated using a normal-phase HPLC column having a totally porous silica stationary phase and using classical reversed-phase eluents. Mass spectrometry in multiple reaction monitoring mode was used for the analysis, providing high selectivity and sensitivity. A standard addition calibration model was applied for quantitation using only one isotope-labeled internal standard for all amino acids. Five calibration points were used for quantitation, and the method was successfully validated. The slopes of the calibration curves of the individual amino acids in parallel measurements were found to be similar. Since the measured slopes were reproducible, one serum sample could represent every series of serum samples of a given day. The method was tested on human serum samples and adequate results were obtained. This new method can be easily applied in clinical laboratories.

Hungarian adaptation of an international swallowing screening method / Egy nemzetközi nyelészavarszurési módszer hazai adaptálása

Introduction: Swallowing disorders caused by stroke can affect half of the cases in the acute phase. The guidelines for nutrition therapy for stroke patients recommend several screening methods for swallowing disorders. The Gugging Swallowing Screen (GUSS) is one of the most widely used ones but has not been available in Hungarian until now. Objective: Adaptation and validation of the GUSS to Hungarian in acute stroke patients (GUSS-H). Method: Our research design was two-phased: for the adaptation, a five-step protocol was composed according to international guidelines. The second phase was the validation of the GUSS-H. For external validity, data from patients (n = 31) were compared to the reference values of the fiberoptic endoscopic evaluation of swallowing (FEES) for both dysphagia and aspiration risk. Internal validity was obtained by comparing data from two independent evaluators (n = 20). Results: According to the FEES results, dysphagia prevalence was 45%, aspiration prevalence was 32.3% in our sample. Inter-rater reliability was strong on both GUSS-H scores and severity of dysphagia (𝜅 = 0.899, p<0.001; 𝜅 = 0.801, p<0.001). The diagnostic accuracy of the test showed great results for both the risk of dysphagia and aspiration (sensitivity: 93%, 90%; specificity: 65%, 57%; positive predictive value: 68%, 50%; negative predictive value: 92%, 92%). Discussion: Compared to the original GUSS and other bedside screenings, GUSS-H performed better than average in terms of sensitivity and negative predictive value. It could predict the risk of dysphagia and aspiration, make recommendations for instrumental evaluation and dysphagia diet. Conclusion: Swallowing screening is one of the first steps of nutritional therapy for acute stroke patients which needs an interdisciplinary setting. With our study, GUSS-H is now available to Hungarian professionals.

Aspirin Mediates Its Antitumoral Effect Through Inhibiting PTTG1 in Pituitary Adenoma.

CONTEXT: DNA demethylation and inhibitory effects of aspirin on pituitary cell proliferation have been demonstrated. OBJECTIVE: Our aim was to clarify the molecular mechanisms behind the aspirin-related effects in pituitary cells. METHODS: DNA methylome and whole transcriptome profile were investigated in RC-4B/C and GH3 pituitary cell lines upon aspirin treatment. Effects of aspirin and a demethylation agent, decitabine, were further tested in vitro. PTTG1 expression in 41 human PitNET samples and whole genome gene and protein expression data of 76 PitNET and 34 control samples (available in Gene Expression Omnibus) were evaluated. RESULTS: Aspirin induced global DNA demethylation and consequential transcriptome changes. Overexpression of Tet enzymes and their cofactor Uhrf2 were identified behind the increase of 5-hydroxymethylcytosine (5hmC). Besides cell cycle, proliferation, and migration effects that were validated by functional experiments, aspirin increased Tp53 activity through p53 acetylation and decreased E2f1 activity. Among the p53 controlled genes, Pttg1 and its interacting partners were downregulated upon aspirin treatment by inhibiting Pttg1 promoter activity. 5hmC positively correlated with Tet1-3 and Tp53 expression, and negatively correlated with Pttg1 expression, which was reinforced by the effect of decitabine. Additionally, high overlap (20.15%) was found between aspirin-regulated genes and dysregulated genes in PitNET tissue samples. CONCLUSION: A novel regulatory network has been revealed, in which aspirin regulated global demethylation, Tp53 activity, and Pttg1 expression along with decreased cell proliferation and migration. 5hmC, a novel tissue biomarker in PitNET, indicated aspirin antitumoral effect in vitro as well. Our findings suggest the potential beneficial effect of aspirin in PitNET.

Medical nutrition therapy of stroke patients with dysphagia ­ 2021 / A dysphagia táplálásterápiája stroke-betegek esetében - 2021.

Összefoglaló. A neurológiai betegek körében a dysphagia elofordulása gyakori, és több oka van. Az utóbbi évek kutatásai a közvetlen neurológiai kórokok (beleértve a gyakori stroke) szerepét is részletesen feltárták. Felismerték az ún. néma aspiráció jelentoségét: ez gyakran áll az (aspirációs) pneumonia hátterében, amely sokszor halálos szövodmény lehet. Az ún. poststroke pneumonia fogalma gyökeresen más értelmezésbe helyezte a stroke-ot követo tüdogyulladások megítélését, jellegzetessége alapján egyértelmuen a stroke közvetlen cerebralis hatásaként alakul ki. Egyértelmuvé vált a nyelészavar korai felismerésének és ellátásának szükségessége. A stroke-betegek megfelelo tápláltsági állapota az eredményes rehabilitációnak, a szövodményszám csökkentésének, a rövidebb kórházi kezelésnek, az alacsonyabb mortalitásnak a záloga. A dysphagia a betegség kimenetelének független elorejelzoje lehet, különösen az elso három hónapban. A nyelészavar malnutritióval, kiszáradással és a kórházi tartózkodás hosszabb idotartamával jár együtt, emeli a gyógyszerköltségeket. A stroke-beteg ellátásának egyik elso eleme a dysphagia szurése. Táplálásterápiára akkor szorul a stroke-beteg, amikor magas a kóros tápláltsági állapot kialakulásának kockázata, és per os táplálással nem fedezheto biztonságosan a megfelelo energia-, tápanyag- és folyadékbevitel. A táplálásterápia módját, eszközeit, az energia- és tápanyagbeviteli célértékeket az orvos határozza meg, az alapbetegség súlyosságától, a társbetegségektol és a laborértékektol függoen. Az étrend minden esetben individuális és progresszív, azaz alkalmazkodik a beteg állapotához és annak változásához. A dietetikus feladata a megfelelo diéta összeállítása mellett a beteg, a hozzátartozó és a kezeloszemélyzet oktatása, az állapot követése, a beteg tápláltsági állapotának, tápanyagbeviteli értékeinek gyakori elemzése, szükség esetén tápszerek ajánlása. Orv Hetil. 2021; 162(40): 1601-1609. Summary. Among neurological patients, the incidence of dysphagia is common and has several causes. Research in recent years has explored the role of direct neurological pathogens (including frequent strokes). The frequency of 'silent aspiration', which often underlies (aspirational) pneumonia and can be a fatal complication, has been recently discovered. The concept of 'post-stroke pneumonia' has drastically changed the assessment of post-stroke pneumonia. Based on its characteristics, it clearly develops as a direct cerebral effect of stroke. The need for early detection and early care of swallowing disorder has become clear. Adequate nutritional status of stroke patients is the key to successful rehabilitation, reduction of complications, shorter hospitalization, and lower mortality. Dysphagia can be an independent predictor of disease outcome, especially in the first three months. Swallowing disorder is associated with malnutrition, dehydration and longer lengths of hospital stay, increasing drug costs. One of the first elements in the care of a stroke patient is screening for dysphagia. The stroke patient needs nutritional therapy when the risk for abnormal nutritional condition is high or if the condition is already present, or when oral nutrition does not safely cover adequate energy, nutrient and fluid intake. The method and means of nutritional therapy, the goals of energy and nutrient intake are determined by the doctor, depending on the severity of the underlying disease, comorbidities and laboratory values.The diet is individual and progressive in each case. The dietitian's task is not only to compile a proper diet, but also to educate the patients and relatives. The dietitian is responsible for monitoring the patient's nutritional status. Orv Hetil. 2021; 162(40): 1601-1609.

Three-component synthesis, utilization and biological activity of phosphinoyl-functionalized isoindolinones.

A new method for the synthesis of 3-oxoisoindolin-1-ylphosphine oxides bearing same or different substituents on the phosphorus atom is described. The one-pot three-component reaction of 2-formylbenzoic acid, primary amines and achiral or P-stereogenic secondary phosphine oxides provided the target compounds under catalyst-free, mild conditions and for short reaction times. The deoxygenation of a 3-oxoisoindolin-1-ylphosphine oxide was also studied, and the phosphine obtained could be converted to a sulphide and to a platinum complex. The crystal structures of a selected phosphine oxide and the corresponding platinum species were investigated by X-ray diffraction analysis. The biological activity, such as in vitro cytotoxicity on different cell lines and antibacterial activity of the 3-oxoisoindolin-1-ylphosphine oxides was also investigated. Based on the IC50 values obtained, several derivatives showed moderate activity against the HL-60 cell line and two compounds containing 3,5-dimethylphenyl groups on the phosphorus atom showed promising activity against Bacillus subtilis bacteria.

Improved Inhibitory and Absorption, Distribution, Metabolism, Excretion, and Toxicology (ADMET) Properties of Blebbistatin Derivatives Indicate That Blebbistatin Scaffold Is Ideal for drug Development Targeting Myosin-2.

Blebbistatin, para-nitroblebbistatin (NBleb), and para-aminoblebbistatin (AmBleb) are highly useful tool compounds as they selectively inhibit the ATPase activity of myosin-2 family proteins. Despite the medical importance of the myosin-2 family as drug targets, chemical optimization has not yet provided a promising lead for drug development because previous structure-activity-relationship studies were limited to a single myosin-2 isoform. Here we evaluated the potential of blebbistatin scaffold for drug development and found that D-ring substitutions can fine-tune isoform specificity, absorption-distribution-metabolism-excretion, and toxicological properties. We defined the inhibitory properties of NBleb and AmBleb on seven different myosin-2 isoforms, which revealed an unexpected potential for isoform specific inhibition. We also found that NBleb metabolizes six times slower than blebbistatin and AmBleb in rats, whereas AmBleb metabolizes two times slower than blebbistatin and NBleb in human, and that AmBleb accumulates in muscle tissues. Moreover, mutagenicity was also greatly reduced in case of AmBleb. These results demonstrate that small substitutions have beneficial functional and pharmacological consequences, which highlight the potential of the blebbistatin scaffold for drug development targeting myosin-2 family proteins and delineate a route for defining the chemical properties of further derivatives to be developed. SIGNIFICANCE STATEMENT: Small substitutions on the blebbistatin scaffold have beneficial functional and pharmacological consequences, highlighting their potential in drug development targeting myosin-2 family proteins.

Study on the Microwave-Assisted Batch and Continuous Flow Synthesis of N-Alkyl-Isoindolin-1-One-3-Phosphonates by a Special Kabachnik-Fields Condensation.

A simple and efficient microwave (MW)-assisted method was elaborated for the catalyst-free synthesis of isoindolin-1-one-3-phosphonates by the three-component condensation of 2-formylbenzoic acid, aliphatic primary amines and various dialkyl phosphites. The batch and the continuous flow reactions were optimized in respect of the temperature, the reaction time and the molar ratio of the starting materials. To evaluate the potential of MW irradiation, comparative thermal experiments were also carried out. In order to obtain "real time" information about the condensation, the special Kabachnik-Fields reaction of 2-formylbenzoic acid, butylamine and diethyl phosphite was monitored by in situ FT-IR spectroscopy. The novel title compounds could be prepared in high yields at low temperature under a short reaction time. A suitable method could also be developed for the preparation of the isoindolin-1-one-3-phosphonates at a "few g" scale by using a continuous flow MW reactor.

Synthesis and In Vitro Cytotoxicity and Antibacterial Activity of Novel 1,2,3-Triazol-5-yl-phosphonates.

Novel 1,2,3-triazol-5-yl-phosphonates were prepared by the copper(I)-catalyzed domino reaction of phenylacetylene, organic azides and dialkyl phosphites. The process was optimized on the synthesis of the dibutyl (1-benzyl-4-phenyl-1H-1,2,3-triazol-5-yl)phosphonate in respect of the catalyst, the base and the solvent, as well as of the reaction parameters (molar ratio of the starting materials, atmosphere, temperature and reaction time). The method elaborated could be applied to a range of organic azides and dialkyl phosphites, which confirmed the large scope and the functional group tolerance. The in vitro cytotoxicity on different cell lines and the antibacterial activity of the synthesized 1,2,3-triazol-5-yl-phosphonates was explored. According to the IC50 values determined, only modest antibacterial effect was detected, while some derivatives showed moderate activity against human promyelocytic leukemia HL-60 cells.

Synthesis of (1,2,3-triazol-4-yl)methyl Phosphinates and (1,2,3-Triazol-4-yl)methyl Phosphates by Copper-Catalyzed Azide-Alkyne Cycloaddition.

An efficient and practical method was developed for the synthesis of new (1,2,3triazol4yl)methyl phosphinates and (1,2,3-triazol-4-yl)methyl phosphates by the copper(I)catalyzed azide-alkyne cycloaddition (CuAAC) of organic azides and prop-2-ynyl phosphinate or prop-2-ynyl phosphate. The synthesis of (1benzyl-1H-1,2,3-triazol-4-yl)methyl diphenylphosphinate was optimized with respect to the reaction parameters, such as the temperature, reaction time, and catalyst loading. The approach was applied to a range of organic azides, which confirmed the wide scope and the substituent tolerance of the process. The method elaborated represents a novel approach for the synthesis of the target compounds.

Simultaneous determination of ten nonsteroidal anti-inflammatory drugs from drinking water, surface water and wastewater using micro UHPLC-MS/MS with on-line SPE system.

A simple, accurate and sensitive micro UHPLC-MS/MS method was developed and validated for the simultaneous determination of 10 nonsteroidal anti-inflammatory drugs (NSAIDs) from different environmental matrices. The micro LC ‒ on-line SPE method described in this study allowed to determine the selected drugs at ultra-trace levels without the most commonly used complex off-line SPE sample preparation procedures. The presented method is capable of reaching satisfactory low LOQ values with analysing the sample directly after being diluted with water. In order to attain high sensitivity, mass spectrometry was carefully optimized for the analysis of the drugs. Fenoprofen, flurbiprofen and naproxen were found to produce CO2 loss during ionization, forming intense [M-H-CO2]- ions instead of [M-H]-. All the other compounds were analyzed through their [M+H]+ and [M-H]- ions. Effect of mobile phase pH on ionization was also studied. Lower pH resulted in higher ion intensities. For this reason, a reversed phase chromatographic separation was applied at pH 3.1 with formic acid at concentration of 0.01%. Matrix effects have been evaluated during validation and sample dilution was optimized focusing on the lowest achievable LOQ values. Analytes were determined from drinking water directly, from surface water and wastewater following dilution with purified water by 2 : 8 (v/v) and 1 : 9 (v/v), respectively. Finally, the method was applied to real sample analysis.

Simultaneous determination of thirteen different steroid hormones using micro UHPLC-MS/MS with on-line SPE system.

Ultratrace analysis of sample components requires excellent analytical performance in terms of limits of quantitation (LOQ). Micro UHPLC coupled to sensitive tandem mass spectrometry provides state of the art solution for such analytical problems. Using on-line SPE with column switching on a micro UHPLC-MS/MS system allowed to decrease LOQ without any complex sample preparation protocol. The presented method is capable of reaching satisfactory low LOQ values for analysis of thirteen different steroid molecules from human plasma without the most commonly used off-line SPE or compound derivatization. Steroids were determined by using two simple sample preparation methods, based on lower and higher plasma steroid concentrations. In the first method, higher analyte concentrations were directly determined after protein precipitation with methanol. The organic phase obtained from the precipitation was diluted with water and directly injected into the LC-MS system. In the second method, low steroid levels were determined by concentrating the organic phase after steroid extraction. In this case, analytes were extracted with ethyl acetate and reconstituted in 90/10 water/acetonitrile following evaporation to dryness. This step provided much lower LOQs, outperforming previously published values. The method has been validated and subsequently applied to clinical laboratory measurement.

Pushing quantitation limits in micro UHPLC-MS/MS analysis of steroid hormones by sample dilution using high volume injection.

Ultratrace analysis of sample components requires excellent analytical performance in terms of limits of quantitation (LoQ). Micro UHPLC coupling with sensitive tandem mass spectrometry provides state of the art solutions for such analytical problems. Decreased column volume in micro LC limits the injectable sample volume. However, if analyte concentration is extremely low, it might be necessary to inject high sample volumes. This is particularly critical for strong sample solvents and weakly retained analytes, which are often the case when preparing biological samples (protein precipitation, sample extraction, etc.). In that case, high injection volumes may cause band broadening, peak distortion or even elution in dead volume. In this study, we evaluated possibilities of high volume injection onto microbore RP-LC columns, when sample solvent is diluted. The presented micro RP-LC-MS/MS method was optimized for the analysis of steroid hormones from human plasma after protein precipitation with organic solvents. A proper sample dilution procedure helps to increase the injection volume without compromising peak shapes. Finally, due to increased injection volume, the limit of quantitation can be decreased by a factor of 2-5, depending on the analytes and the experimental conditions.

Adverse events in a newborn on valproate therapy due to loss-of-function mutations in CYP2C9.

An increased risk of valproate-induced toxicity has been reported in children, particularly in those younger than 2 years of age. Significant variations in valproate pharmacokinetics and shifts in the metabolic pathways towards CYP2C9-dependent metabolism seem to play some role in the age-related differences in the incidence of adverse events. We present the case of a premature patient with moderate hemorrhage in the subependymal region (grade II - intraventricular hemorrhage without ventricular dilatation), several myoclonic episodes in her right upper arm (series of jerks lasting milliseconds), and epileptiform abnormalities on the EEG (localized spike-and-wave in the left frontal region with preserved background activity who was treated with valproate. Serious side effects, consisting of bone marrow depression, hyperammonemia, and serum alkaline phosphatase elevation, were observed seventeen days after the beginning of valproate therapy. The toxic symptoms were likely the consequence of a reduced ability to metabolize valproate. The patient was demonstrated to carry two loss-of-function mutations in CYP2C9 (CYP2C9*3/*3) resulting in exaggerated blood concentrations of valproate. The present case highlights the importance of assaying inborn errors in CYP2C9 gene in pediatric patients to avoid valproate-evoked serious side effects.