The neural correlates of mental fatigue and reward processing: A task-based fMRI study.

Increasing time spent on the task (i.e., the time-on-task (ToT) effect) often results in mental fatigue. Typical effects of ToT are decreasing levels of task-related motivation and the deterioration of cognitive performance. However, a massive body of research indicates that the detrimental effects can be reversed by extrinsic motivators, for example, providing rewards to fatigued participants. Although several attempts have been made to identify brain areas involved in mental fatigue and related reward processing, the neural correlates are still less understood. In this study, we used the psychomotor vigilance task to induce mental fatigue and blood oxygen-level-dependent functional magnetic resonance imaging to investigate the neural correlates of the ToT effect and the reward effect (i.e., providing extra monetary reward after fatigue induction) in a healthy young sample. Our results were interpreted in a recently proposed neurocognitive framework. The activation of the right middle frontal gyrus, right insula and right anterior cingulate gyrus decreased as fatigue emerged and the cognitive performance dropped. However, after providing an extra reward, the cognitive performance, as well as activation of these areas, increased. Moreover, the activation levels of all of the mentioned areas were negatively associated with reaction times. Our results confirm that the middle frontal gyrus, insula and anterior cingulate cortex play crucial roles in cost-benefit evaluations, a potential background mechanism underlying fatigue, as suggested by the neurocognitive framework.

Gray Matter Changes Following Mild COVID-19: An MR Morphometric Study in Healthy Young People.

BACKGROUND: Although COVID-19 is primarily an acute respiratory infection, 5%-40% of patients develop late and prolonged symptoms with frequent neurological complaints, known as long COVID syndrome. The presentation of the disease suggests that COVID infection may cause functional and/or morphological central nervous system alterations, but studies published in the literature report contradictory findings. PURPOSE: To investigate the chronic effects of COVID-19 on cerebral grey matter in a group of young patients without comorbidities, with mild course of COVID infection and no medical complaints at the time of examination. STUDY TYPE: Prospective. POPULATION: Thirty-eight young (age = 26.6 ± 5.0 years; male/female = 14/24), adult participants who recovered from mild COVID infection without a history of clinical long COVID and 37 healthy control subjects (age = 25.9 ± 2.8 years; male/female = 14/23). FIELD STRENGTH/SEQUENCE: Three Tesla, 3D T1-weighted magnetization-prepared rapid gradient-echo, 2D T2-weighted turbo spin-echo. ASSESSMENT: MRI-based morphometry and volumetry along with neuropsychological testing and self-assessed questionnaire. STATISTICAL TESTS: Fisher's exact test, Mann-Whitney U-test, and multiple linear regression analyses were used to assess differences between COVID and healthy control groups. P < 0.05 was used as cutoff for significance. RESULTS: In the COVID group, significantly lower bilateral mean cortical thickness (left/right-hemisphere: 2.51 ± 0.06 mm vs. 2.56 ± 0.07 mm, η2 p = 0.102/2.50 ± 0.06 mm vs. 2.54 ± 0.07 mm, η2 p = 0.101), lower subcortical gray matter (57881 ± 3998 mm3 vs. 60470 ± 5211 mm3 , η2 p = 0.100) and lower right olfactory bulb volume (52.28 ± 13.55 mm3 vs. 60.98 ± 15.8 mm3 , η2 p = 0.078) were found. In patients with moderate to severe anosmia, cortical thickness was significantly lower bilaterally, as compared to patients without olfactory function loss (left/right-hemisphere: 2.50 ± 0.06 mm vs. 2.56 ± 0.05 mm, η2 = 0.173/2.49 ± 0.06 mm vs. 2.55 ± 0.05 mm, η2 = 0.189). Using further exploratory analysis, significantly reduced cortical thickness was detected locally in the right lateral orbitofrontal cortex in the COVID group (2.53 ± 0.10 mm vs. 2.60 ± 0.09 mm, η2 p = 0.112). DATA CONCLUSION: Even without any subjective or objective neurological complaints at the time of the MR scan, subjects in the COVID group showed gray matter alterations in cortical thickness and subcortical gray matter volume. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY: Stage 3.

Co-Channel Compatibility Analysis of IMT Networks and Digital Terrestrial Television Broadcasting in the Frequency Range 470-694 MHz Based on Monte Carlo Simulation.

According to Resolution 235 adopted during the World Radiocommunication Conference 2015 (WRC-15) and the preliminary agenda item 1.5 of WRC-2023, the International Telecommunication Union Radiocommunication Sector (ITU-R) has been entrusted to carry out sharing and compatibility studies between broadcasting and mobile services in the frequency range 470-694 MHz in Region 1. This study specifically focuses on evaluating electromagnetic compatibility in potential co-channel sharing scenarios between Digital Terrestrial Television Broadcasting (DTTB) and International Mobile Telecommunications (IMT) systems within the 470-694 MHz frequency band that may arise in the foreseeable future. To assess the conditions for coexistence, a Monte Carlo simulation method implemented through the SEAMCAT software version 5.4.2 is employed, examining six potential interference scenarios. The simulation results yield the minimum coordination distance required between IMT and DTTB services in the 470-694 MHz band based on the protection criteria to ensure harmonious coexistence while maintaining satisfactory performance. Furthermore, the study investigates the impact of various factors such as transmitter power, antenna heights, coverage radius, antenna discrimination, and antenna tilt angle on the separation distances. The focus lies primarily on critical interference scenarios across neighboring countries' borders. The simulation outcomes confirm that sharing the same frequency band between IMT and DTTB networks would result in significant mutual interference. Nevertheless, carefully analyzing diverse parameters and assumptions helped provide recommendations to reduce the required separation distances. These findings are valuable for broadcasters, mobile operators, and regulators in establishing the technical coexistence conditions for DTTB and IMT in the new band.

Arabidopsis RETINOBLASTOMA RELATED directly regulates DNA damage responses through functions beyond cell cycle control.

The rapidly proliferating cells in plant meristems must be protected from genome damage. Here, we show that the regulatory role of the Arabidopsis RETINOBLASTOMA RELATED (RBR) in cell proliferation can be separated from a novel function in safeguarding genome integrity. Upon DNA damage, RBR and its binding partner E2FA are recruited to heterochromatic γH2AX-labelled DNA damage foci in an ATM- and ATR-dependent manner. These γH2AX-labelled DNA lesions are more dispersedly occupied by the conserved repair protein, AtBRCA1, which can also co-localise with RBR foci. RBR and AtBRCA1 physically interact in vitro and in planta Genetic interaction between the RBR-silenced amiRBR and Atbrca1 mutants suggests that RBR and AtBRCA1 may function together in maintaining genome integrity. Together with E2FA, RBR is directly involved in the transcriptional DNA damage response as well as in the cell death pathway that is independent of SOG1, the plant functional analogue of p53. Thus, plant homologs and analogues of major mammalian tumour suppressor proteins form a regulatory network that coordinates cell proliferation with cell and genome integrity.

Provocation and prediction of visual peripersonal neglect-like symptoms in preoperative planning and during awake brain surgery.

Neglect is a severe neuropsychological/neurological deficit that usually develops due to lesions of the posterior inferior parietal area of the right hemisphere and is characterized by a lack of attention to the left side. Our case is a proven right-handed, 30-year-old female patient with a low-grade glioma, which was located in the temporo-opercular region and also in the superior temporal gyrus of the right hemisphere. Upon presurgical planning, the motor, language, and visuospatial functions were mapped. In order to achieve this, the protocol for routine magnetic resonance imaging and navigated transcranial magnetic stimulation has been expanded, accordingly.

Application of Structural Entropy and Spatial Filling Factor in Colonoscopy Image Classification.

For finding colorectal polyps the standard method relies on the techniques and devices of colonoscopy and the medical expertise of the gastroenterologist. In case of images acquired through colonoscopes the automatic segmentation of the polyps from their environment (i.e., from the bowel wall) is an essential task within computer aided diagnosis system development. As the number of the publicly available polyp images in various databases is still rather limited, it is important to develop metaheuristic methods, such as fuzzy inference methods, along with the deep learning algorithms to improve and validate detection and classification techniques. In the present manuscript firstly a fuzzy rule set is generated and validated. The former process is based on a statistical approach and makes use of histograms of the antecedents. Secondly, a method for selecting relevant antecedent variables is presented. The selection is based on the comparision of the histograms computed from the measured values for the training set. Then the inclusion of the Rényi-entropy-based structural entropy and the spatial filling factor into the set of input variables is proposed and assessed. The beneficial effect of including the mentioned structural entropy of the entropies from the hue and saturation (H and S) colour channels resulted in 65% true positive and 60% true negative rate of the classification for an advantageously selected set of antecedents when working with HSV images.

A novel family of expression vectors with multiple affinity tags for wheat germ cell-free protein expression.

BACKGROUND: Cell-free protein expression has become a widely used alternative of in vivo, cell-based systems in functional and structural studies of proteins. The wheat germ-based method outstands from the commercially available eukaryotic in vitro translation systems by its flexibility, high translation efficiency and success rate of properly folded eukaryotic protein synthesis. The original T7 promoter containing pEU3-NII vector was improved previously by addition of a ligation-independent cloning site, His6- and GST-tags, and a TEV protease cleavage site to facilitate the creation of recombinant plasmids, permit affinity purification, and enable production of purified, tag-free target proteins, respectively. RESULTS: Here, we describe a further development of pEU3-NII vector by inserting the rare-cutting, NotI restriction enzyme cleavage site to simplify vector linearization step prior to in vitro transcription. Additionally, His12, FLAG, and Halo affinity tag coding vectors have been created to increase detection sensitivity, specificity of interaction studies, and provide covalently linkable ligands for pull-down assays, respectively. Finally, the presented GST-His6, and GST-biotin double-tagging vectors could broaden the range of possibilities of protein-protein interaction studies. CONCLUSIONS: The new generation of pEU3-NII vector family allows a more rapid production of translationally active mRNA and wheat germ cell-free expression of target proteins with a wide variety of affinity tags thus enables designing flexible and diverse experimental arrangement for in vitro studies of proteins.

Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system.

BACKGROUND: Elaboration of the epigenetic regulation of chromatin is a long-standing aim in molecular and cellular biology. Hence, there is a great demand for the development of in vitro methods to reconstitute chromatin that can be used directly for biochemical assays. The widely used wheat germ cell-free protein expression method provides broad applications to investigate the function and structure of eukaryotic proteins. Such advantages, including high translation efficiency, flexibility, and possible automatization, are beneficial for achieving native-like chromatin substrates for in vitro studies. RESULTS: We describe a novel, single-step in vitro chromatin assembly method by using the wheat germ cell-free protein synthesis. We demonstrated that both Drosophila and human chromatins can be reconstituted in the course of the in vitro translation of core histones by the addition of chromatin assembly factors, circular plasmid, and topoisomerase I in an ATP-dependent manner. Drosophila chromatin assembly was performed in 4 h at 26 °C, in the presence of premixed mRNAs encoding the core histones, dAcf1/dISWI chromatin remodeling complex, and nucleosome assembly protein, dNAP1. Similarly, the human chromatin was assembled by co-expressing the human core histones with Drosophila chromatin remodeling factor, dISWI, and chromatin chaperone, dNLP, for 6 h at 26 °C. The presence of reconstituted chromatin was monitored by DNA supercoiling assay, also the regular spacing of nucleosomes was assessed by Micrococcal nuclease assay. Furthermore, Drosophila linker histone H1-containing chromatin was reconstituted, affirming that the in vitro assembled chromatin is suitable for downstream applications. CONCLUSIONS: The method described in this study allows the assembly of Drosophila and human chromatins, possibly in native-like form, by using a wheat germ cell-free protein expression. Although both chromatins were reconstituted successfully, there were unexpected differences with respect to the required ratio of histone-coding mRNAs and the reaction time. Overall, our new in vitro chromatin reconstitution method will aid to characterize the unrevealed structure, function, and regulation of chromatin dynamics.

Ultrasonic Sensor Fusion Inverse Algorithm for Visually Impaired Aiding Applications.

Depth mapping can be carried out by ultrasound measuring devices using the time of flight method. Ultrasound measurements are favorable in such environments, where the light or radio frequency measurements can not be applied due to the noise level, calculation complexity, reaction time, size and price of the device, accuracy or electromagnetic compatibility. It is also usual to apply and fusion ultrasound sensors with other types of sensors to increase the precision and reliability. In the case of visually impaired people, an echolocation based aid for determining the distance and the direction of obstacles in the surroundings can improve the life quality by giving the possibility to move alone and individually in unlearnt or rapidly changing environments. In the following considerations, a model system is presented which can provide rather reliable position and distance to multiple objects. The mathematical model based on the time of flight method with a correction: it uses the measured analog sensor signals, which represent the probability of the presence of an obstacle. The device consists of multiple receivers, but only one source. The sensor fusion algorithm for this setup and the results of indoor experiments are presented. The mathematical model allows the usage, processing, and fusion of the signals of up to an infinite number of sensors. In addition, the positions of the sensors can be arbitrary, and the mathematical model does not restrict them to be placed in regular formations.

Influence of hemispheric white matter lesions and migraine characteristics on cortical thickness and volume.

BACKGROUND/AIM: Migraine-related intracerebral white matter lesions (WMLs) are likely to be microvascular in nature and can be found in all hemispheric lobes. The aim of this study was to investigate migraine patients with or without WMLs to see the effects of these tissue damages on cortical thickness and volume. The role of migraine characteristics (duration of headache, attack frequency, estimated lifetime attack number, aura) was also tested. METHODS: As study participants, 161 female migraine patients (63 with aura; 52 with WMLs) and 40 age-matched healthy female subjects were enrolled in the study. None of the included migraine patients' headache or aura (where present) was unilaterally side-locked. Patients and controls were all right handed. Except for migraine, patients were free of any medical comorbidity. Cortical reconstruction and segmentation were performed on the 3D T1-weighted images using Freesurfer 5.3 image analysis suite. The automatic cortical parcellation was based on Freesurfer's Desikan-Killiany-Tourville atlas, which has 31 cortical regions per hemisphere. The segmented regions were divided into five lobes (frontal, parietal, temporal, occipital, insula). Since the left and right differences in lobar and insular volumes/thicknesses were not different among our groups, volume and cortical thickness were calculated for corresponding bilateral lobes. RESULTS: There was no significant difference in age between the whole migraine and the control groups. Migraineurs with WMLs (L+ patients) were significantly older than lesion-free (L-) patients (P = 0.0003) and controls (P = 0.018). Disease duration (P = 0.003), the total number of migraine attacks (P = 0.022) and the rate of aura (P = 0.0003) were significantly higher in L+ patients than in L- patients. Cortical thickness and volume measurements of lobes were not statistically different between the three groups (L+, L-, control). Age showed a significant negative association with both thickness and volume in each examined lobe (P < 0.001). Intracranial volume (ICV) showed a significant positive association with all regional volumes (P < 0.001). There were no significant group*age, group*ICV, or age*ICV interactions. None of the migraine characteristics were selected by stepwise linear regression as significant predictors of cortical thickness or volume. Only age (for both thickness and volume) and ICV (for volume) were identified as significant predictors (P < 0.001). When the L + group was divided into two subgroups by median split of total and lobar lesion number and volume, the cortical measures did not show any significant difference between the groups with low vs. high lesion number/volume by stepwise linear regression. CONCLUSIONS: In a female migraine group, we found that the WMLs and clinical migraine characteristics have no effect on cortical thickness and volume of bilateral lobes. Lobar cortical thicknesses were equivalent within the range of ±0.1 mm. Only age and ICV proved to be significant predictors; the former for both cortical thickness and volume, while the latter for cortical volume.

On Structural Entropy and Spatial Filling Factor Analysis of Colonoscopy Pictures.

Colonoscopy is the standard device for diagnosing colorectal cancer, which develops from little lesions on the bowel wall called polyps. The Rényi entropies-based structural entropy and spatial filling factor are two scale- and resolution-independent quantities that characterize the shape of a probability distribution with the help of characteristic curves of the structural entropy-spatial filling factor map. This alternative definition of structural entropy is easy to calculate, independent of the image resolution, and does not require the calculation of neighbor statistics, unlike the other graph-based structural entropies.The distant goal of this study was to help computer aided diagnosis in finding colorectal polyps by making the Rényi entropy based structural entropy more understood. The direct goal was to determine characteristic curves that can differentiate between polyps and other structure on the picture. After analyzing the distribution of colonoscopy picture color channels, the typical structures were modeled with simple geometrical functions and the structural entropy-spatial filling factor characteristic curves were determined for these model structures for various parameter sets. A colonoscopy image analying method, i.e., the line- or column-wise scanning of the picture, was also tested, with satisfactory matching of the characteristic curve and the image.

Characterization of auxin transporter PIN6 plasma membrane targeting reveals a function for PIN6 in plant bolting.

Auxin gradients are sustained by series of influx and efflux carriers whose subcellular localization is sensitive to both exogenous and endogenous factors. Recently the localization of the Arabidopsis thaliana auxin efflux carrier PIN-FORMED (PIN) 6 was reported to be tissue-specific and regulated through unknown mechanisms. Here, we used genetic, molecular and pharmacological approaches to characterize the molecular mechanism(s) controlling the subcellular localization of PIN6. PIN6 localizes to endomembrane domains in tissues with low PIN6 expression levels such as roots, but localizes at the plasma membrane (PM) in tissues with increased PIN6 expression such as the inflorescence stem and nectary glands. We provide evidence that this dual localization is controlled by PIN6 phosphorylation and demonstrate that PIN6 is phosphorylated by mitogen-activated protein kinases (MAPKs) MPK4 and MPK6. The analysis of transgenic plants expressing PIN6 at PM or in endomembrane domains reveals that PIN6 subcellular localization is critical for Arabidopsis inflorescence stem elongation post-flowering (bolting). In line with a role for PIN6 in plant bolting, inflorescence stems elongate faster in pin6 mutant plants than in wild-type plants. We propose that PIN6 subcellular localization is under the control of developmental signals acting on tissue-specific determinants controlling PIN6-expression levels and PIN6 phosphorylation.

Kinase-Associated Phosphoisoform Assay: a novel candidate-based method to detect specific kinase-substrate phosphorylation interactions in vivo.

BACKGROUND: Protein kinases are important components of signalling pathways, and kinomes have remarkably expanded in plants. Yet, our knowledge of kinase substrates in plants is scarce, partly because tools to analyse protein phosphorylation dynamically are limited. Here we describe Kinase-Associated Phosphoisoform Assay, a flexible experimental method for directed experiments to study specific kinase-substrate interactions in vivo. The concept is based on the differential phosphoisoform distribution of candidate substrates transiently expressed with or without co-expression of activated kinases. Phosphorylation status of epitope-tagged proteins is subsequently detected by high-resolution capillary isoelectric focusing coupled with nanofluidic immunoassay, which is capable of detecting subtle changes in isoform distribution. RESULTS: The concept is validated by showing phosphorylation of the known mitogen-activated protein kinase (MAPK) substrate, ACS6, by MPK6. Next, we demonstrate that two transcription factors, WUS and AP2, both of which are shown to be master regulators of plant development by extensive genetic studies, exist in multiple isoforms in plant cells and are phosphorylated by activated MAPKs. CONCLUSION: As plant development flexibly responds to environmental conditions, phosphorylation of developmental regulators by environmentally-activated kinases may participate in linking external cues to developmental regulation. As a counterpart of advances in unbiased screening methods to identify potential protein kinase substrates, such as phosphoproteomics and computational predictions, our results expand the candidate-based experimental toolkit for kinase research and provide an alternative in vivo approach to existing in vitro methodologies.

Biexponential diffusion alterations in the normal-appearing white matter of glioma patients might indicate the presence of global vasogenic edema.

PURPOSE: To investigate normal-appearing white matter (NAWM) microstructure of glioma patients with biexponential diffusion analysis in order to reveal the nature of diffusion abnormalities and to assess whether they are region-specific or global. MATERIALS AND METHODS: Twenty-four newly diagnosed glioma patients (grade II-IV) and 24 matched control subjects underwent diffusion-weighted imaging at 3T. Diffusion parameters were calculated using monoexponential and biexponential models. Apparent diffusion coefficient (ADC) values were measured in the entire NAWM of the hemisphere contralateral and ipsilateral to the tumor. In the contralateral NAWM, regional ADC values were assessed in the frontal, parietal, occipital, and temporal NAWM. RESULTS: ADCmono and ADCfast were significantly higher than control values in all investigated regions except the temporal NAWM (P < 0.04). ADCslow was significantly increased in the total contralateral, frontal, and parietal NAWM (P < 0.03), while pslow was decreased in both total hemispheric NAWM and the parietal NAWM of glioma patients compared to controls (P < 0.04). ADCmono , ADCfast , ADCslow , and pslow were significantly different among the NAWM of the four lobes of the contralateral hemisphere in both groups (P < 0.0001), and these regional differences were similar in patients and controls (P > 0.05). Hemispheric ADCmono and pslow differences were different between groups (P < 0.05). CONCLUSION: Globally altered diffusion parameters suggest the presence of global vasogenic edema in the NAWM of glioma patients, which is further supported by the finding that regional differences in patients follow those found in controls. Alternatively, some tumor infiltration might contribute to diffusion abnormalities in the NAWM, especially in the tumor-affected hemisphere. J. Magn. Reson. Imaging 2016;44:633-641.

Increased diffusion in the normal appearing white matter of brain tumor patients: is this just tumor infiltration?

Altered diffusion in the normal appearing white matter (NAWM) of glioma patients has been explained by tumor infiltration. The goal of the present study was to test this explanation indirectly by examining whether these alterations were also present in the contralateral NAWM of non-infiltrative tumors like meningiomas; and to search for other possible reasons for this abnormality. Twenty-seven patients with histologically verified glioma (grade II-IV), 22 meningioma patients and two groups of age- and sex-matched healthy controls underwent diffusion weighted imaging (DWI) on a 3T MR. All patients were examined before treatment. Apparent diffusion coefficient (ADC) values were calculated in the entire NAWM of the hemisphere contralateral to the tumor. ADC values of the NAWM were compared between groups with Mann-Whitney U-test and multiple linear regression. The relations of ADC in NAWM to glioma grade and to tumor volume were also investigated. ADC values of the contralateral NAWM were significantly higher in both glioma and meningioma patients compared to controls (P = 0.0006 and 0.0099, respectively). ADC value was higher in the NAWM of high grade gliomas than in low grade gliomas (P = 0.0181) and in healthy control subjects (P = 0.0003). ADC did not depend on tumor volume in any of the patient groups. Elevated ADC in the NAWM of both glioma and meningioma patients might indicate that the effect of infiltrating tumor cells is not the only reason for the alteration as it has been previously suggested. Although the role of mass effect was not proved, other mechanisms might also contribute to ADC elevation.

Age at onset and seizure frequency affect white matter diffusion coefficient in patients with mesial temporal lobe epilepsy.

In mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS), structural abnormalities are present not only in the hippocampus but also in the white matter with ipsilateral predominance. Although the timing of epilepsy onset is commonly associated with clinical and semiological dissimilarities, limited data exist regarding white matter diffusion changes with respect to age at epilepsy onset. The aim of this study was to investigate diffusion changes in the white matter of patients with unilateral MTLE-HS with respect to clinical parameters and to compare them with an age- and sex-matched healthy control group. Apparent diffusion coefficients (ADCs) were derived using monoexponential approaches from 22 (11 early and 11 late age at onset) patients with unilateral MTLE-HS and 22 age- and sex-matched control subjects after acquiring diffusion-weighted images on a 3T MRI system. Data were analyzed using two-tailed t-tests and multiple linear regression models. In the group with early onset MTLE-HS, ADC was significantly elevated in the ipsilateral hemispheric (p=0.04) and temporal lobe white matter (p=0.01) compared with that in controls. These differences were not detectable in late onset MTLE-HS patients. Apparent diffusion coefficient of the group with early onset MTLE-HS was negatively related to age at epilepsy onset in the ipsilateral hemispheric white matter (p=0.03) and the uncinate fasciculus (p=0.03), while in patients with late onset MTLE-HS, ADC was no longer dependent on age at epilepsy onset itself but rather on the seizure frequency in the ipsilateral uncinate fasciculus (p=0.03). Such diffusivity pattern has been associated with chronic white matter degeneration, reflecting myelin loss and higher extracellular volume which are more pronounced in the frontotemporal regions and also depend on clinical features. In the group with early onset MTLE-HS, the timing of epilepsy seems to be the major cause of white matter abnormalities while in late onset disease, it has a secondary role in provoking diffusion changes.

Activation of AtMPK9 through autophosphorylation that makes it independent of the canonical MAPK cascades.

Mitogen-activated protein kinases (MAPKs) are part of conserved signal transduction modules in eukaryotes that are typically organized into three-tiered kinase cascades. The activation of MAPKs in these pathways is fully dependent on the bisphosphorylation of the TXY motif in the T-loop by the pertinent dual-specificity MAPK kinases (MAPKKs). The Arabidopsis mitogen-activated protein kinase 9 (AtMPK9) is a member of an atypical class of MAPKs. Representatives of this MAPK family have a TDY phosphoacceptor site, a long C-terminal extension and lack the common MAPKK-binding docking motif. In the present paper, we describe multiple in vitro and in vivo data showing that AtMPK9 is activated independently of any upstream MAPKKs but rather is activated through autophosphorylation. We mapped the autophosphorylation sites by MS to the TDY motif and to the C-terminal regulatory extension. We mutated the phosphoacceptor sites on the TDY, which confirmed the requirement for bisphorylation at this site for full kinase activity. Next, we demonstrated that the kinase-inactive mutant form of AtMPK9 is not trans-phosphorylated on the TDY site when mixed with an active AtMPK9, implying that the mechanism of the autocatalytic phosphorylation is intramolecular. Furthermore, we show that in vivo AtMPK9 is activated by salt and is regulated by okadaic acid-sensitive phosphatases. We conclude that the plant AtMPK9 shows similarities to the mammalian atypical MAPKs, such as extracellular-signal-regulated kinase (ERK) 7/8, in terms of an MAPKK-independent activation mechanism.

THE RELATIONSHIP BETWEEN SERUM CHOLESTEROL AND VERBAL MEMORY MAY BE INFLUENCED BY BODY MASS INDEX (BMI) IN YOUNG HEALTHY WOMEN.

The objective of this study was to examine the relationship between verbal memory and total cholesterol (TC) levels related to body mass index (BMI) in healthy young women. Verbal memory was assessed using the Rey Auditory-Verbal Learning Test (RAVLT) while total serum cholesterol was measured by enzymatic colorimetric test. In order to analyze the potential significance of BMI subjects were divided into three groups according to their calculated BMI percentile values. No significant correlation was found when assessing the group as a whole. However a remarkable pattern of correlation emerged when assessing the BMI groups separately: a close-to-significant positive correlation was found for total learning score and TC in the low BMI group, no correlation emerged in the medium BMI group while a strong inverse correlation was found in the high BMI group. These findings indicate that the relationship between verbal memory and serum TC level is also influenced by BMI.

The Arabidopsis mitogen-activated protein kinase 6 is associated with γ-tubulin on microtubules, phosphorylates EB1c and maintains spindle orientation under nitrosative stress.

Stress-activated plant mitogen-activated protein (MAP) kinase pathways play roles in growth adaptation to the environment by modulating cell division through cytoskeletal regulation, but the mechanisms are poorly understood. We performed protein interaction and phosphorylation experiments with cytoskeletal proteins, mass spectrometric identification of MPK6 complexes and immunofluorescence analyses of the microtubular cytoskeleton of mitotic cells using wild-type, mpk6-2 mutant and plants overexpressing the MAP kinase-inactivating phosphatase, AP2C3. We showed that MPK6 interacted with γ-tubulin and co-sedimented with plant microtubules polymerized in vitro. It was the active form of MAP kinase that was enriched with microtubules and followed similar dynamics to γ-tubulin, moving from poles to midzone during the anaphase-to-telophase transition. We found a novel substrate for MPK6, the microtubule plus end protein, EB1c. The mpk6-2 mutant was sensitive to 3-nitro-l-tyrosine (NO2 -Tyr) treatment with respect to mitotic abnormalities, and root cells overexpressing AP2C3 showed defects in chromosome segregation and spindle orientation. Our data suggest that the active form of MAP kinase interacts with γ-tubulin on specific subsets of mitotic microtubules during late mitosis. MPK6 phosphorylates EB1c, but not EB1a, and has a role in maintaining regular planes of cell division under stress conditions.

Quantitative assessment of brain iron by R2* relaxometry in patients with cervical dystonia.

BACKGROUND: The pathophysiology of cervical dystonia is poorly understood. Increased brain iron deposition has been described in different movement disorders. Our aim was to investigate brain iron content in patients with cervical dystonia, using R2* relaxation rate, a validated MRI marker of brain iron level. METHODS: Twelve female patients with primary focal cervical dystonia (mean age: 45.4 ± 8.0 years) and 12 age-matched healthy female subjects (mean age: 45.0 ± 8.0 years) underwent 3T MRI to obtain regional R2* relaxation rates of the thalamus, caudate nucleus, putamen, and globus pallidus (GP). Regions of interest were delineated automatically on T1-weighted MRIs. RESULTS: R2* values in the putamen were positively correlated with age. Patients with cervical dystonia showed elevated R2* values in the GP. CONCLUSIONS: This pilot study provides the first quantitative support for increased brain iron deposition in cervical dystonia. Further studies are needed to explore the implications of this finding.