The implications of DNA methylation for amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a complex and serious neurodegenerative disorder that develops in consequence of the progressive loss of the upper and lower motor neurons. Cases of ALS are classified as sporadic (sALS), or familial (fALS). Over 90% of cases are sALS, while roughly 10% are related to inherited genetic mutations (fALS). Approximately 70% of the genetic mutations that contribute to fALS have been identified. On the other hand, the majority of the sALS cases have an undetermined genetic contributor and few mutations have been described, despite the advanced genetic analysis methods. Also, several factors contribute to the onset and progression of ALS. Numerous lines of evidence indicate that epigenetic changes are linked to aging, as well as neurodegenerative disorders, such as ALS. In most cases, they act as the heritable regulation of transcription by DNA methylation, histone modification and expression of noncoding RNAs. Mechanisms involving aberrant DNA methylation could be relevant to human ALS pathobiology and therapeutic targeting. Despite advances in research to find factors associated with ALS and more effective treatments, this disease remains complex and has low patient survival. Here, we provide a narrative review of the role of DNA methylation for this complex neurodegenerative disorder.

Neurochemical changes in basal ganglia affect time perception in parkinsonians.

BACKGROUND: Parkinson's disease is described as resulting from dopaminergic cells progressive degeneration, specifically in the substantia nigra pars compacta that influence the voluntary movements control, decision making and time perception. AIM: This review had a goal to update the relation between time perception and Parkinson's Disease. METHODOLOGY: We used the PRISMA methodology for this investigation built guided for subjects dopaminergic dysfunction in the time judgment, pharmacological models with levodopa and new studies on the time perception in Parkinson's Disease. We researched on databases Scielo, Pubmed / Medline and ISI Web of Knowledge on August 2017 and repeated in September 2017 and February 2018 using terms and associations relevant for obtaining articles in English about the aspects neurobiology incorporated in time perception. No publication status or restriction of publication date was imposed, but we used as exclusion criteria: dissertations, book reviews, conferences or editorial work. RESULTS/DISCUSSION: We have demonstrated that the time cognitive processes are underlying to performance in cognitive tasks and that many are the brain areas and functions involved and the modulators in the time perception performance. CONCLUSIONS: The influence of dopaminergic on Parkinson's Disease is an important research tool in Neuroscience while allowing for the search for clarifications regarding behavioral phenotypes of Parkinson's disease patients and to study the areas of the brain that are involved in the dopaminergic circuit and their integration with the time perception mechanisms.

Genetic influence alters the brain synchronism in perception and timing.

BACKGROUND: Studies at the molecular level aim to integrate genetic and neurobiological data to provide an increasingly detailed understanding of phenotypes related to the ability in time perception. MAIN TEXT: This study suggests that the polymorphisms genetic SLC6A4 5-HTTLPR, 5HTR2A T102C, DRD2/ANKK1-Taq1A, SLC6A3 3'-UTR VNTR, COMT Val158Met, CLOCK genes and GABRB2 A/C as modification factor at neurochemical levels associated with several neurofunctional aspects, modifying the circadian rhythm and built-in cognitive functions in the timing. We conducted a literature review with 102 studies that met inclusion criteria to synthesize findings on genetic polymorphisms and their influence on the timing. CONCLUSION: The findings suggest an association of genetic polymorphisms on behavioral aspects related in timing. However, order to confirm the paradigm of association in the timing as a function of the molecular level, still need to be addressed future research.

Bioimpedance patterns and bioelectrical impedance vector analysis (BIVA) of road cyclists.

Bioelectrical impedance vector-analysis (BIVA) describes cell-mass, cell function and hydration status of an individual or a group. The goal of the present investigation was to provide bioelectrical impedance data for 525 male road cyclists (155 professionals, 79 elite, 59 elite-youth, and 232 amateurs) at the time of their optimal performance level. Data were plotted on the resistance-reactance (R-Xc) graph to characterize cyclists group vectors using BIVA. Compared to the general male population, the mean vector position of the road cyclists indicates a higher body cell mass (BCM) and phase angle (p<0.001). The vector position of the high-performance, compared to the amateur cyclists showed similar patterns with higher BCM and phase angles and higher reactance values for the high-performance athletes (p<0.001). The bio-impedance data were used to calculate the 50%, 75%, and 95% tolerance ellipses of each group of cyclists. The characteristic vector positions of the road cyclists indicate normal hydration and greater muscle mass and function of the high-performance cyclists compared to amateur cyclists and the normal population. The cyclists specific tolerance ellipses, particularly the high-performance cyclists might be used for classifying a cyclist according to the individual vector position and to define target vector regions for lower level cyclists.

The dopaminergic system dynamic in the time perception: a review of the evidence.

Dopaminergic system plays a key role in perception, which is an important executive function of the brain. Modulation in dopaminergic system forms an important biochemical underpinning of neural mechanisms of time perception in a very wide range, from milliseconds to seconds to longer daily rhythms. Distinct types of temporal experience are poorly understood, and the relationship between processing of different intervals by the brain has received little attention. A comprehensive understanding of interval timing functions should be sought within a wider context of temporal processing, involving genetic aspects, pharmacological models, cognitive aspects, motor control and the neurological diseases with impaired dopaminergic system. Particularly, an unexplored question is whether the role of dopamine in interval timing can be integrated with the role of dopamine in non-interval timing temporal components. In this review, we explore a wider perspective of dopaminergic system, involving genetic polymorphisms, pharmacological models, executive functions and neurological diseases on the time perception. We conclude that the dopaminergic system has great participation in impact on time perception and neurobiological basis of the executive functions and neurological diseases.

Proprioceptive neuromuscular facilitation increases alpha absolute power in the dorsolateral prefrontal cortex and superior parietal cortex.

The physiotherapist's clinical practice includes proprioceptive neuromuscular facilitation (PNF), which is a treatment concept that accelerates the response of neuromuscular mechanisms through spiral and diagonal movements. The adaptations that occur in the nervous system following PNF are still poorly described in the literature. Thus, this study had a goal to investigate the electrophysiological changes in the fronto-parietal circuit during PNF and movement in sagittal and diagonal patterns. This study included 30 female participants, who were divided into three groups (control, PNF, and flexion groups). Electroencephalogram measurements were determined before and after tasks were performed by each group. For the statistical analysis, a two-way ANOVA was performed for the factors group and time. Interactions between the two factors were investigated using a one-way ANOVA. A value of p < 0.004 was considered significant. The results showed an increase in alpha absolute power in the left dorsolateral prefrontal cortex and upper left parietal cortex of the PNF group, suggesting these areas work together to execute a motor action. The PNF group showed a greater alpha absolute power compared with the other groups, indicating a specific cortical demand for planning and attention, reinforcing its use for the rehabilitation of individuals.

Effects of vestibular rehabilitation in the elderly: a systematic review.

BACKGROUND: Aging is characterized by gradual physiological changes in body systems. Changes in the vestibular system can occur and cause dizziness, vertigo and imbalance, symptoms that are common in the elderly. Vestibular rehabilitation is a therapeutic resource that has been widely used to improve this condition. AIMS: To complete a systematic review of the effects of vestibular rehabilitation on the elderly. METHODS: A search for relevant publications was conducted in SCIELO, PUBMED, MEDLINE, COCHRANE and LILACS databases. Clinical trials and cohort studies that were written in the English language and published over the course of the last 10 years were selected. The methodological quality of the studies was assessed using the PEDro scale. A critical analysis of the studies was composed. RESULTS: Eight studies that involved subjects who were over the age of 60 were selected for inclusion in the systematic review. The most common vestibular dysfunction identified was complaints about dizziness and imbalance. The Dizziness Handicap Inventory was the most frequently used assessment instrument, and the treatment protocol that prevailed was that suggested by Cawthorne and Cooksey. DISCUSSION: The PEDro scale showed that only one article was of an acceptable methodological quality and presented satisfactory outcome measures. This was due, in part, to a lack of a hidden randomization, masking of the subject, evaluators and therapists, and lack of outcome measures, which can reduce the quality of the evidence presented in this study. CONCLUSION: Clinical trials indicate that vestibular rehabilitation represents an effective means of treating elderly patients with vestibular disorders; however, evidence of its effectiveness remains lacking.

Intra-observer reliability in three-dimensional kinematic analysis of sacroiliac joint mobility.

[Purpose] Physical therapists, osteopathic practitioners, and chiropractors often perform manual tests to evaluate sacroiliac joint (SIJ) mobility. However, the available evidence demonstrates an absence of reliability in these tests and in investigations with kinematic analysis. The aim of this study was to verify the three-dimensional kinematic reliability in SIJ movement measurements. [Subjects] This cross-sectional study analyzed 24 healthy males, aged between 18 and 35 years. [Methods] Three-dimensional kinematic analysis was performed for measurements of posterior superior iliac displacement and greater trochanter (femur) displacement during hip flexion movement in an orthostatic position. The distance variations were measured from a reference point in 3 blocks. The intra-observer reliability was compared with the mean of three 3 blocks using the interclass correlation coefficient (ICC) and a 99% significance level. [Results] The measurements indicated a strong correlation among blocks: ICC = 0.94 for right side SIJ and ICC = 0.91 for left side SIJ. The mean displacement between the reference points was 7.7 mm on the right side and 8.5 mm on the left side. [Conclusion] Our results indicate that three-dimensional kinematic analysis can be used for SIJ mobility analyses. New studies should be performed for subjects with SIJ dysfunction to verify the effectiveness of this method.

Comparison of the effects of hamstring stretching using proprioceptive neuromuscular facilitation with prior application of cryotherapy or ultrasound therapy.

[Purpose] Stretching using proprioceptive neuromuscular facilitation involve physiological reflex mechanisms through submaximal contraction of agonists which activate Golgi organ, promoting the relaxation reflex. The aim of this study was to evaluate the effects of proprioceptive neuromuscular facilitation alone and with prior application of cryotherapy and thermotherapy on hamstring stretching. [Subjects and Methods] The sample comprised of 32 young subjects with hamstring retraction of the right limb. The subjects were randomly allocated to four groups: the control, flexibility PNF, flexibility PNF associated with cryotherapy, flexibility PNF in association with ultrasound therapy. [Results] After 12 stretching sessions, experimental groups showed significant improvements compared to the control group. Moreover, we did not find any significant differences among the experimental groups indicating PNF stretching alone elicits similar results to PNF stretching with prior administration of cryotherapy or thermotherapy. [Conclusion] PNF without other therapy may be a more practical and less expensive choice for clinical care.

Electrocortical Analysis of Patients with Intercostobrachial Pain Treated with TENS after Breast Cancer Surgery.

[Purpose] Among the physical therapeutic procedures to decrease pain, there is transcutaneous electrical neural stimulation (TENS). There is no consensus about its efficacy for oncological patients, especially for post-mastectomy pain and eletrocortical changes in somatosensory areas. The aim of this study was to analyze acute electrocortical changes after TENS treatment of patients with intercostobrachial post mastectomy pain. [Subjects] Eighteen patients were divided into acupuncture (A) and burst (B) group. [Methods] In this pre and post-intervention study each group was measured for EEG analysis in absulte power in alpha band (8-14 Hz). Outcomes variables were the alpha waveband in the sensorymotor cortex and pain pre-and-post TENS intervention. Data were analyzed using ANOVA to compare times (rest, 10 and 15 min), group and electrodes. Pain was analyzed using percentual pain evaluation (PPE) in both groups. [Results] Outcomes indicate main effects of time and electrodes because of slow (8-10 Hz) and fast alpha (10-12 Hz) wavebands decreased. PPE reduced 88.4% in A and 66.3% in G. [Conclusion] TENS promoted electrical modification in the parietal region and a decrease in pain. Future studies should investigate other wave must be proposed for other bands and use different methods of EEG analysis to elucidate the actual mechanisms behind the efficacy of TENS treatment.

Comparative analysis electroencephalographic of alpha, Beta and gamma bands of a healthy individual and one with hemiparesis.

[Purpose] The study analyzed the electroencephalographic (EEG) data of the central cortical areas, during execution of the motor gestures of feeding, activation of the system of mirror neurons, and imagery between a right hemiparetic volunteer (RHV) and a healthy volunteer (HV). [Subjects and Methods] The volunteers' EEG data were recorded with their eyes open for 4 minutes while they performed five experimental tasks. [Results] The alpha band, absolute power value of HV was lower than that of RHV. In the beta band, during the practice condition, there was an increase in the magnitude of the absolute power value of HV at T3, possibly because T3 is representative of secondary motor areas that work with cortical neurons related to planning and organizing sequence of movements performed by the hands. The gamma band is related to the state of preparation for movement and memory. The results of this study indicate that there was increased activation of the gamma frequency band of HV. [Conclusion] The findings of this study have revealed the changes in pattern characteristics of each band which may be associated with the brain injury of the hemiparetic patient.

Changes in Cortical Activity During Real and Imagined Movements: an ERP Study.

This study aims to compare the topographic distribution of cortical activation between real and imagined movement through event-related potential (ERP). We are specifically interested in identifying, the topographic distribution of activated areas, the intensity of activated areas, and the temporal occurrence of these activations on preparation and motor response phases. Twelve healthy and right handed subjects were instructed to perform a task under real and imagery conditions. The task was performed simultaneously to electroencephalographic (EEG) recording. When compared the conditions, we found a statistically significant difference in favor of real condition revealed by performing an unpaired t-test with multiple corrections of Bonferroni, demonstrating negative activity on electrode C3 and positive activity on the electrode C4 only in motor response phase. These findings revealed similar functional connections established during real and imagery conditions, suggesting that there are common neural substrate and similar properties of functional integration shared by conditions.

Differential Diagnosis or Etiology: A Case Report on Amyotrophic Lateral Sclerosis-like Neuropathy Associated with HIV Infection.

BACKGROUND: Retroviruses are described as a risk factor for chronic neuropathy. However, it is still unknown if they can work as amyotrophic lateral sclerosis triggers. Over the years, some cases of this association have been described with heterogenous disclosures. CASE REPRESENTATION: This study aimed to report a case of HIV and ALS-like neuropathy and briefly discuss peculiarities of clinical aspects, such as physiopathology and treatment options. The patient underwent neurological examination associated with blood tests, electromyography, analysis of cerebrospinal fluid, and imaging studies. DISCUSSION: A non-systematic review was performed in major databases regarding the topic. The case presented mixed upper and lower motor neuron signs and was framed as a probable case of ALS following the present criteria. CONCLUSION: After a short follow-up and viral load cleansing, neurological stabilization was achieved.

Proinflammatory Cytokine Expression in Apical Periodontitis from Diabetic Patients.

Diabetes mellitus has been regarded as a condition capable of influencing the evolution of periapical lesions. Therefore, this study evaluated the immunoexpression of IL-1ß, TNF-α, and IL-17 in apical periodontitis from type 2 diabetic patients through immunohistochemistry. Twenty-six periapical lesions were selected, and the images obtained through immunohistochemistry reactions were analyzed. The statistical significance was set at p < 0.05. IL-1ß expression was considered focal (54%), weak to moderate (38%), and strong (8%) in diabetics, and focal (46%), weak to moderate (31%), and strong (23%), in controls. TNF-α was focal (85%) and weak to moderate (15%) in diabetics, and focal (92%) and weak to moderate (8%), in controls. IL-17 was focal (8%), weak to moderate (46%), and strong (46%) in diabetics, and focal (62%), weak to moderate (8%), and strong (30%), in controls. The quantitative analysis revealed greater expression of IL-17, with a significant difference between IL-17 × IL-1ß × TNF-α(p=0.0009) in the diabetic group. The cytokines IL-1ß and TNF-α did not express statistical differences between the tested groups. The IL-17 showed higher immunoexpression in the diabetic group (p=0.047), which may suggest higher bone resorption activity in chronic apical periodontitis in this group of patients.

Differences between hemispheres and in saccade latency regarding volleyball athletes and non-athletes during saccadic eye movements: an analysis using EEG.

BACKGROUND: The saccadic eye movement is responsible for providing focus to a visual object of interest to the retina. In sports like volleyball, identifying relevant targets quickly is essential to a masterful performance. The training improves cortical regions underlying saccadic action, enabling more automated processing in athletes. OBJECTIVE: We investigated changes in the latency during the saccadic eye movement and the absolute theta power on the frontal and prefrontal cortices during the execution of the saccadic eye movement task in volleyball athletes and non-athletes. We hypothesized that the saccade latency and theta power would be lower due to training and perceptual-cognitive enhancement in volleyball players. METHODS: We recruited 30 healthy volunteers: 15 volleyball athletes (11 men and 4 women; mean age: 15.08 ± 1.06 years) and 15 non-athletes (5 men and 10 women; mean age: 18.00 ± 1.46 years). All tasks were performed simultaneously with electroencephalography signal recording. RESULTS: The latency of the saccadic eye movement presented a significant difference between the groups; a shorter time was observed among the athletes, associated with the players' superiority in terms of attention level. During the experiment, the athletes observed a decrease in absolute theta power compared to non-athletes on the electrodes of each frontal and prefrontal area. CONCLUSION: In the present study, we observed the behavior of reaction time and absolute theta power in athletes and non-athletes during a saccadic movement task. Our findings corroborate the premise of cognitive improvement, mainly due to the reduction of saccadic latency and lower beta power, validating the neural efficiency hypothesis.

ANTECEDENTES: O movimento ocular sacádico é responsável por dar foco a um objeto visual de interesse para a retina. Em esportes como o vôlei, identificar alvos relevantes o mais rápido possível é essencial para se ter um desempenho magistral. O treinamento melhora as regiões corticais subjacentes à ação sacádica, e permite um processamento mais automatizado em atletas. OBJETIVO: Investigamos as mudanças na latência durante o movimento ocular sacádico e a potência teta absoluta nos córtices frontal e pré-frontal durante a execução da tarefa de movimento ocular sacádico em atletas e não atletas de voleibol. Nossa hipótese é a de que a latência sacádica e a potência teta seriam menores em atletas devido ao treinamento e ao aprimoramento perceptivo-cognitivo em jogadores de voleibol. MéTODOS: Ao todo, 30 voluntários saudáveis foram recrutados para este estudo: 15 atletas de voleibol (11 homens e 4 mulheres; idade média: 15,08 ± 1,06 anos) e 15 não atletas (5 homens e 10 mulheres; idade média: 18,00 ± 1,46 anos). Todas as tarefas foram realizadas simultaneamente com o registro do sinal eletroencefalográfico. RESULTADOS: O resultado da latência do movimento ocular sacádico apresentou diferença significativa entre os grupos, sendo observado menor tempo entre os atletas, associado à superioridade dos jogadores em termos de nível de atenção. Durante o experimento, nos eletrodos de cada área frontal e pré-frontal, observou-se uma diminuição na potência teta absoluta nos atletas em comparação aos não atletas. CONCLUSãO: Neste estudo, observou-se o comportamento do tempo de reação e da potência teta absoluta em atletas e não atletas durante uma tarefa de movimento sacádico. Nossos achados corroboram a premissa de melhora cognitiva, principalmente pela redução da latência sacádica e menor potência beta, o que valida a hipótese de eficiência neural.

Clinical efficacy of a xenogeneic collagen matrix in augmenting keratinized mucosa around implants: a randomized controlled prospective clinical trial.

AIM: The aim of this controlled randomized clinical trial was to evaluate the efficacy of a xenogeneic collagen matrix (CM) to augment the keratinized tissue around implants supporting prosthetic restorations at 6 months when compared with the standard treatment, the connective tissue autograft, CTG). MATERIALS AND METHODS: This randomized longitudinal parallel controlled clinical trial studied 24 patients with at least one location with minimal keratinized tissue (≤1 mm). MAIN OUTCOME MEASURE: The 6-month width of keratinized tissue. As secondary outcomes the esthetic outlook, the maintenance of peri-implant mucosal health and the patient morbidity were assessed pre-operatively and 1, 3, and 6 months post-operatively. RESULTS: At 6 months, Group CTG attained a mean width of keratinized tissue of 2.75 (1.5) mm, while the corresponding figure in Group CM was 2.8 (0.4) mm, the inter-group differences not being statistically significant. The surgical procedure in both groups did not alter significantly the mucosal health in the affected abutments. There was a similar esthetic result and significant increase in the vestibular depth in both groups as a result of the surgery. In the CM group it changed from 2.2 (3.3) to 5.1 (2.5) mm at 6 months. The patients treated with the CM referred less pain, needed less pain medication, and the surgical time was shorter, although these differences were not statistically significant when compared with the CTG group. CONCLUSIONS: These results prove that this new CM was as effective and predictable as the CTG for attaining a band of keratinized tissue.

Response letter: Neuropsychiatric presentation of Covid-19-related encephalitis: Case report.

Covid-19-related encephalitis is a heterogeneous syndrome characterized by a combination of clinical, laboratory, and imaging features related to inflammation of the brain, where the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is presumably the causative agent. We reported a case of Covid-19-related encephalitis presenting with neuropsychiatric symptoms, including intense agitation. Reverse-transcriptase polymerase-chain-reaction in cerebrospinal fluid was positive for SARS-CoV-2. Our case expands the literature about neurologic manifestations of Covid-19 and emphasizes the possibility of prominent behavioral symptoms as the initial manifestation.

Neuropsychiatric presentation of Covid-19-related encephalitis: Case report.

Covid-19-related encephalitis is a heterogeneous syndrome characterized by a combination of clinical, laboratory, and imaging features related to inflammation of the brain, where the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is presumably the causative agent. We reported a case of Covid-19-related encephalitis presenting with neuropsychiatric symptoms, including intense agitation. Reverse-transcriptase polymerase-chain-reaction in cerebrospinal fluid was positive for SARS-CoV-2. Our case expands the literature about neurologic manifestations of Covid-19 and emphasizes the possibility of prominent behavioral symptoms as the initial manifestation.

Association between Body Composition and Dysphagia in Patients with Amyotrophic Lateral Sclerosis.

BACKGROUND: The influence of changes in body composition on swallowing in patients with Amyotrophic Lateral Sclerosis (ALS) is unknown. Understanding the interrelation between body compartments and dysphagia may establish specific treatments related to both nutritional aspects as to myofunctional ones designed to delay swallowing loss. AIM: The aim of the study was to evaluate the relationship between body composition and dysphagia during the course of the disease. METHODS: The protocol of this study included assessments carried out quarterly for one year and included: analysis of body composition by multi-frequency segmental bioimpedance, nutritional diagnosis, maximum strength test of the tongue and lips, analysis of swallowing using the Functional Oral Intake Score (FOIS) and Swallowing Rating Scale of the American Speech-Language-Hearing Association (ASHA), speech intelligibility and analysis of disease severity. To measure the degree of relationship between quantitative variables, Spearman's correlation was used. RESULTS: Thirty-four patients were evaluated, 28 Spinal Group and 6 Bulbar Group. The results did not show any significant differences in the analysis of body composition between the groups. Positive associations were found between body compartments and swallowing analysis. The phase angle showed a strong correlation between the FOIS scales (r = 0.74, p < 0.01), ASHA (r = 0.77, p < 0.01) and tongue (r = 0.66, p < 0.01). CONCLUSIONS: Changes in body compartments were related to swallowing functionality and speech intelligibility in ALS patients, emphasizing the importance of analyzing body compartments for decision making by the interdisciplinary team. Although these preliminary data were collected in a small sample size, they serve to motivate future studies in this area.

Repetitive transcranial magnetic stimulation changes cognitive/motor tasks performance: An absolute alpha and beta power study.

The voluntary movement demands integration between cognitive and motor functions. During the initial stages of motor learning until mastery of a new motor task, and during a demanding task that is not automatic, cognitive and motor functions can be perceived as independent from each other. Areas used for actually performing motor tasks are essentially the same used by Motor Imagery (MI). The main objective of this study was to investigate inhibition effects on cognitive functions of motor skills induced by low-frequency (1 Hz) Repetitive Transcranial Magnetic Stimulation (rTMS) at the sensory-motor integration site (Cz). In particular, the goal was to examine absolute alpha and beta power changes on frontal regions during Execution, Action observation, and Motor Imagery of finger movement tasks. Eleven healthy, right-handed volunteers of both sexes (5 males, 6 females; mean age 28 ± 5 years), with no history of psychiatric or neurological disorders, participated in the experiment. The execution task consisted of the subject flexing and extending the index finger. The action observation task involved watching a video of the same movement. The motor imagery task was imagining the flexion and extension of the index finger movement. After performing the tasks randomly, subjects were submitted to 15 min of low-frequency rTMS and performed the tasks again. All tasks were executed simultaneously with EEG signals recording. Our results demonstrated a significant interaction between rTMS and the three tasks in almost all analyzed regions showing that rTMS can affect the frontal region regarding Execution, Action observation, and Motor Imagery tasks.