Altered brain activity for phonological manipulation in dyslexic Japanese children.

Because of unique linguistic characteristics, the prevalence rate of developmental dyslexia is relatively low in the Japanese language. Paradoxically, Japanese children have serious difficulty analysing phonological processes when they have dyslexia. Neurobiological deficits in Japanese dyslexia remain unclear and need to be identified, and may lead to better understanding of the commonality and diversity in the disorder among different linguistic systems. The present study investigated brain activity that underlies deficits in phonological awareness in Japanese dyslexic children using functional magnetic resonance imaging. We developed and conducted a phonological manipulation task to extract phonological processing skills and to minimize the influence of auditory working memory on healthy adults, typically developing children, and dyslexic children. Current experiments revealed that several brain regions participated in manipulating the phonological information including left inferior and middle frontal gyrus, left superior temporal gyrus, and bilateral basal ganglia. Moreover, dyslexic children showed altered activity in two brain regions. They showed hyperactivity in the basal ganglia compared with the two other groups, which reflects inefficient phonological processing. Hypoactivity in the left superior temporal gyrus was also found, suggesting difficulty in composing and processing phonological information. The altered brain activity shares similarity with those of dyslexic children in countries speaking alphabetical languages, but disparity also occurs between these two populations. These are initial findings concerning the neurobiological impairments in dyslexic Japanese children.

[Early detection of specific reading disorder through response to intervention:a cohort study].

OBJECTIVE: The present study examined the effectiveness of response to intervention (RTI) for the early detection and intervention for specific reading disorder. METHOD: RTIs were applied to seventy-seven first graders (36 boys and 41 girls) for the early detection and intervention of specific reading disorders. The outcomes were examined when the children reached the third grade. RESULTS: Reading difficulties were detected in four children at the beginning of the first grade. Decoding training and vocabulary learning were conducted with these children as interventions for reading difficulties. Three children exhibited improvements in reading difficulties at the end of the first grade. The fourth child responded poorly and was diagnosed with specific reading disorder. The assessment of outcomes at the third grade showed that this child still had reading difficulties, whereas no other child displayed symptoms for being diagnosed with specific reading disorder. CONCLUSION: RTI is effective for the early detection and intervention of specific reading disorder.

Long-Term Resveratrol Intake for Cognitive and Cerebral Blood Flow Impairment in Carotid Artery Stenosis/Occlusion.

BACKGROUND AND PURPOSE: Carotid artery stenosis or occlusion (CASO) is a causative disease of vascular cognitive impairment (VCI) attributed to cerebral hypoperfusion, even without the development of symptomatic ischemic stroke. Preclinically, resveratrol has been demonstrated to play an important role in improving cognitive function in rodent CASO models. This study investigated the association between long-term resveratrol intake and improvements in cognitive and cerebral hemodynamic impairments in patients with CASO. METHODS: A retrospective cohort study was conducted on patients with asymptomatic carotid artery stenosis of ≥50% or occlusion who underwent 15O-gas positron emission tomography (15O-gas PET) and neuropsychological tests such as Montreal Cognitive Assessment (MoCA) and Alzheimer's Disease Assessment Scale-Cognitive Subscale 13 (ADAS-Cog) twice between July 2020 and March 2022 allowing >125-day interval. Patients were administered 30 mg/day resveratrol after the first 15O-gas PET and neuropsychological tests were compared with those who were not. RESULTS: A total of 79 patients were enrolled in this study; 36 received resveratrol and 43 did not. Over a mean follow-up of 221.2 and 244.8 days, long-term resveratrol treatment significantly improved visuospatial/executive function (P=0.020) in MoCA, and memory domain (P=0.007) and total score (P=0.019) in ADAS-Cog. Cerebral blood flow demonstrated improvement in the right frontal lobe (P=0.027), left lenticular nucleus (P=0.009), right thalamus (P=0.035), and left thalamus (P=0.010) on 15O-gas PET. No adverse events were reported. CONCLUSION: Long-term daily intake of oral resveratrol may prevent or treat VCI by improving the cerebral blood flow in asymptomatic patients with CASO.

Interlaboratory evaluation of LC-MS-based biomarker assays.

Validation of biomarker assays is crucial for effective drug development and clinical applications. Interlaboratory reproducibility is vital for reliable comparison and combination of data from different centers. This review summarizes interlaboratory studies of quantitative LC-MS-based biomarker assays using reference standards for calibration curves. The following points are discussed: trends in reports, reference and internal standards, evaluation of analytical validation parameters, study sample analysis and normalization of biomarker assay data. Full evaluation of these parameters in interlaboratory studies is limited, necessitating further research. Some reports suggest methods to address variations in biomarker assay data among laboratories, facilitating organized studies and data combination. Method validation across laboratories is crucial for reducing interlaboratory differences and reflecting target biomarker responses.

Measurement of drug concentration in the stomach after intragastric administration of drug solution to healthy volunteers: analysis of intragastric fluid dynamics and drug absorption.

PURPOSE: To evaluate the time-profile of intragastric fluid volume in humans after intragastric administration of drug solution. METHODS: Eight healthy volunteers were intragastrically administered 150 mL of drug solution containing atenolol (non-absorbable marker) and salicylic acid, then, aliquots of gastric fluid (ca. 2 mL) were sampled for 2 h through the catheter. Rate constants for secretion and emptying of the fluid were obtained by fitting the time-course of atenolol concentration to the simple gastric fluid transit model. Absorption of salicylic acid from the stomach was estimated by comparing its gastric concentration with that of atenolol. RESULTS: Kinetic analysis of atenolol concentration in the stomach indicated a rapid emptying of the fluid with an average half-life of 4.2 min. Steady-state intragastric fluid volume in 8 volunteers was estimated as 4-133 mL with an average of 42 mL. Intragastric concentration (normalized by dose) of salicylic acid was always lower than that of atenolol, showing approximately 40% of salicylic acid was absorbed from the stomach before emptying to the intestine. CONCLUSIONS: This study provided valuable information on intragastric fluid dynamics and gastric drug absorption in humans to establish a better in vitro-in vivo correlation in oral drug absorption.

[Efficacy of decoding training for children with difficulty reading hiragana].

OBJECTIVE: The present study aimed to clarify the efficacy of decoding training focusing on the correspondence between written symbols and their readings for children with difficulty reading hiragana (Japanese syllabary). METHODS: Thirty-five children with difficulty reading hiragana were selected from among 367 first-grade elementary school students using a reading aloud test and were then divided into intervention (n=15) and control (n=20) groups. The intervention comprised 5 minutes of decoding training each day for a period of 3 weeks using an original program on a personal computer. Reading time and number of reading errors in the reading aloud test were compared between the groups. RESULTS: The intervention group showed a significant shortening of reading time (F(1,33)=5.40, p<0.05, two-way ANOVA) compared to the control group. However, no significant difference in the number of errors was observed between the two groups. Ten children in the control group who wished to participate in the decoding training were included in an additional study;as a result, improvement of the number of reading errors was observed (t= 2.863, p< 0.05, paired t test), but there was no improvement in reading time. CONCLUSIONS: Decoding training was found to be effective for improving both reading time and reading errors in children with difficulty reading hiragana.

Multi-laboratory evaluation of immunoaffinity LC-MS-based glucagon-like peptide-1 assay.

Background: Although the fit-for-purpose approach has been proposed for biomarker assay validation, practical data should be compiled to facilitate the predetermination of acceptance criteria. Methods: Immunoaffinity LC-MS was used to analyze glucagon-like peptide-1 as a model biomarker in six laboratories. Calibration curve, carryover, parallelism, precision, relative accuracy and processed sample stability were evaluated, and their robustness among laboratories was assessed. The rat glucagon-like peptide-1 concentrations in four blinded samples were also compared. Results: The obtained results and determined concentrations in the blinded samples at all laboratories were similar, with a few exceptions, and robust, despite the difference in optimization techniques among laboratories. Conclusion: The results provide insights into the predefinition of the acceptance criteria of immunoaffinity LC-MS-based biomarker assays.

Highlights of the 13th Japan Bioanalysis Forum Symposium.

Approximately 280 people from pharmaceutical industries, contractors, academic institutions and regulatory authorities attended the 13th Japan Bioanalysis Forum Symposium. The symposium was held via web to prevent the spread of COVID-19 from the 28 February to 2 March 2022. The theme of the symposium was 'All for One Goal', and the event has provided an opportunity for open discussion among researchers with different backgrounds but who share a common goal: "to deliver more effective and safe pharmaceuticals to patients as quickly as possible". The speakers focused on hot topics in bioanalysis, including chromatography, biomarker analysis, cell and gene therapy, COVID-19 and antidrug antibody. This symposium provided a great opportunity for the participants to have meaningful discussions, even though 'on the web' was a limited space.

A perspective for biowaivers of human bioequivalence studies on the basis of the combination of the ratio of AUC to the dose and the biopharmaceutics classification system.

The ratio of AUC to the dose (AUC/dose) was previously found as a parameter that predicts a risk of bioinequivalence of oral drug products. On the basis of the combination of this parameter and the biopharmaceutics classification system (BCS), a perspective for biowaivers of human bioequivalence studies is discussed. Databases of bioequivalence studies using immediate-release solid oral dosage forms were disclosed by 6 Japanese generic pharmaceutical companies, and the number of subjects required for demonstrating bioequivalence between generic and reference products was plotted as a function of AUC/dose for each BCS category. A small variation in the number of subjects was constantly observed in bioequivalence studies using dosage forms containing an identical BCS class 1 or class 3 drug, even though formulations of the generic product differ between companies. The variation was extremely enlarged when the drugs were substituted with BCS class 2 drugs. Rate-determining steps in oral absorption of highly water-soluble BCS class 1 and class 3 drugs are independent of formulations when there is no significant difference in the in vitro dissolution profiles between formulations. The small variation observed for both BCS categories indicates that the number of subjects converges into one value for each drug. Our analysis indicates the appropriateness of biowaiver of bioequivalence studies for immediate-release solid oral dosage forms containing not only BCS class 1 drugs but also class 3 drugs.

[Reading aloud as rehabilitation method for children with dyslexia detected at the first grade in their primary school].

We provided reading aloud instructions to a child who was diagnosed with dyslexia in a regular class of 69 first graders, comprising 33 boys and 36 girls, during a test of reading sentences aloud. The instructions consisted of a 2-step approach, i.e., decoding instruction and vocabulary instruction. First, a decoding instruction, which emphasized an important point in effortless decoding, was presented to the child. Next, a vocabulary instruction, which aimed to facilitate word-form recognition, was provided. We found that, the decoding instruction was effective in decreasing the number of reading errors, and that the vocabulary instruction was effective against reducing the time taken to read aloud.

A multilaboratory validation study of LC/MS biomarker assays for three lysophosphatidylcholines.

Aim: Although the fit-for-purpose approach has been proposed for validation procedures and acceptance criteria for biomarker assays, practical biomarker assays to facilitate clinical application and regulatory documents on biomarker assays remain limited. Materials & methods: We assigned six independent laboratories and selected three lysophosphatidylcholines (LPCs): LPC(16:0), LPC(18:0) and LPC(18:1) as model biomarkers. Using LC-MS, the following key validation parameters were evaluated: calibration curve, carryover, parallelism, precision and relative accuracy and these values were similar among all laboratories. Further, we determined LPC levels in six lots of rat plasma at unknown concentrations and compared them among the laboratories. Conclusion: Our multilaboratory validation and reproducibility data are useful for the development of future biomarker assay validation procedures, as well as regulatory documents.

Highlights of the 12th Japan Bioanalysis Forum Symposium.

Approximately 300 people associated with pharmaceutical industries, contractors, academic institutions and regulatory authorities attended the 12th Japan Bioanalysis Forum Symposium. The webinar was conducted from 9 to 11 March 2021. The theme of the symposium was 'for the next generation', and the event provided 'an opportunity for young researchers in bioanalysis (including students)' and 'an opportunity to discuss new frontiers of bioanalysis'. The speakers focused on hot topics of bioanalysis, including biomarker analysis, patient centric sampling, virtual clinical trials, gene therapy, cancer genome medicine and therapeutic middle molecules. The symposium presented a platform for the discussion of the prospects and challenges facing bioanalysts working in the field of pharmacokinetics. This report presents the key issues discussed.

Smaller insula and inferior frontal volumes in young adults with pervasive developmental disorders.

Enlarged head circumference and increased brain weight have been reported in infants with pervasive developmental disorders (PDD), and volumetric studies suggest that children with PDD have abnormally enlarged brain volumes. However, little is known about brain volume abnormalities in young adults with PDD. We explored gray matter (GM) volume in young adults with PDD. T1-weighted volumetric images were acquired with a 3-T magnetic resonance scanner from 32 males with high-functioning PDD (23.8+/-4.2 years; Full Scale Intelligence Quotient [FSIQ]=101.6+/-15.6) and 40 age-matched normal male control subjects (22.5+/-4.3 years; FSIQ=109.7+/-7.9). Regional GM volumes were compared between the two groups using voxel-based morphometry (VBM) with the Diffeomorphic Anatomical Registration using Exponentiated Lie algebra (DARTEL). Compared with the control group, the high-functioning PDD group showed significantly less GM in the right insula, the right inferior frontal gyrus, and the right inferior parietal lobule. A conservative threshold confirmed considerably smaller volumes in the right insula and inferior frontal gyrus. In these areas, negative correlations were found between Autism Spectrum Quotient scores and GM volume, although no significant correlations were found between each subject's FSIQ and GM volume. No regions showed greater GM volumes in the high-functioning PDD group. The insular cortex, which works as a relay area for multiple neurocognitive systems, may be one of the key regions underlying the complex clinical features of PDD. These smaller GM volumes in high-functioning PDD subjects may reflect the clinical features of PDD itself, rather than FSIQ.

Incidental findings of brain magnetic resonance imaging study in a pediatric cohort in Japan and recommendation for a model management protocol.

BACKGROUND: The increasing use of magnetic resonance imaging (MRI) in brain researches has led to growing concern over incidental findings (IFs). To establish a practical management protocol for IFs, it is useful to know the actual prevalence and problems of IF management. In the present study, we report the prevalence proportion and some handling problems of IFs in healthy Japanese children, and suggest a management protocol from ethical and practical standpoints. METHODS: Between 2006 and 2008, 120 healthy children aged 5-8 years participated in a structural MRI study conducted in a pediatric cohort in Japan. All MRI images were reviewed by a pediatric neurologist, and detected IFs were classified into 4 categories. RESULTS: IFs of all categories were detected in 40 of the 110 participants (36.4%) for whom T2-weighted or 3D-T1-weighted images were available. Findings of sinusitis and/or otitis media were most frequent (26.4%). Excluding these findings, the prevalence of IFs was still 10.9% (12 findings): 9 findings were categorized as "no referral" (8.2%), 2 as "routine referral" (1.8%), 1 as "urgent referral" (0.9%), and 0 as "immediate referral" (0.0%). In "routine referral" category, only one participant was referred for further examinations. CONCLUSIONS: Although the prevalence of IFs was high, the proportion of those requiring further examination was low. This result revealed a fairly high false-positive rate and suggested that evaluating equivocal findings was the most difficult part of IF management. A management protocol needs to include a process to properly assess the clinical importance of findings.

Uremic serum residue decreases SN-38 sensitivity through suppression of organic anion transporter polypeptide 2B1 in LS-180 colon cancer cells.

Pharmacokinetics of SN-38 in patients with end-stage kidney disease (ESKD) is partially varied because of fluctuations in transporters expression and/or function by high protein bound-uremic toxins concentration. The fluctuations may induce variations in anticancer drugs sensitivity to cancer cells. We aimed to clarify the variations in sensitivity of SN-38 to cancer patients with ESKD and investigate this mechanism, by human colon cancer cells exposed to uremic serum residue. LS180 cells were exposed to normal or uremic serum residue (LS/NSR or LS/USR cells) for a month. IC50 values of SN-38 in LS/NSR or LS/USR cells were calculated from viability of each cells treated SN-38. mRNA expression and intracellular SN-38 accumulation was evaluated by RT-PCR and HPLC-fluorescence methods, respectively. The IC50 value in LS/USR cells was higher than that in LS/NSR cells. Organic anion transporter polypeptide (OATP) 2B1 mRNA expression was lower in LS/USR cells than in LS/NSR cells, and SN-38 accumulation in LS/USR cells was lower than that in LS/NSR cells. Only co-treatment baicalin, which is OATP2B1 inhibitor, almost negated the difference in SN-38 accumulation between LS/NSR and LS/USR. Anticancer effects of substrates of OATP2B1, such as SN-38, were reduced in ESKD patients at the same plasma substrate concentration.

Effects of Uremic Serum Residue on OATP1B1- and OATP1B3-Mediated Pravastatin Uptake in OATP-Expressing HEK293 Cells and Human Hepatocytes.

Patients with end-stage renal disease have increased plasma concentrations of statins, which is a risk factor for rhabdomyolysis, as well as elevated levels of uremic toxins (UTs). We investigated the effects of uremic serum residue and UTs on organic anion-transporting peptide (OATP1B1)- and OATP1B3-mediated pravastatin uptake. We evaluated the effects of normal serum residue with four UTs (hippuric acid, 3-carboxy-4-methyl-5-propyl-2-furan propionate, indole-3-acetic acid, and 3-indoxyl sulfate) and uremic serum residue on pravastatin uptake by OATP1B1- or OATP1B3-expressing HEK293 cells. Furthermore, we assessed the contribution of each transporter using cryopreserved human hepatocytes. Uremic serum residue and UTs significantly inhibited OATP1B1-mediated pravastatin uptake. Uremic serum residue accelerated OATP1B3-mediated pravastatin uptake, while UTs had no effect. There was no difference in pravastatin uptake between uremic- and normal serum residue-treated hepatocytes. The results suggest that the effects of uremic serum on pravastatin hepatic uptake may be considered negligible in end-stage renal disease patients.

The 10th Japan Bioanalysis Forum Symposium conference: a report.

The Japan Bioanalysis Forum Symposium was held on 12-14 February 2019 (Yokohama, Japan), in celebration of its 10th anniversary, and over 370 participants from pharmaceutical industries, contractors, academia and regulatory authorities from home and abroad came together in Yokohama. The 3-day symposium particularly aimed to foster collaboration with the scientists surrounding bioanalysts, according to the theme 'Open to the Public.' The symposium also included a broad range of pioneering programs, such as lectures by speakers from DMPK/metabolomics fields, discussions of future bioanalysis and poster presentations by publicly offered presenters as well as the regular ones we had organized. This report summarizes the major topics as a conference report.

Reading ability and phonological awareness in Japanese children with dyslexia.

UNLABELLED: In alphabetic languages, the deficit of the phonological awareness is considered as the core deficit in developmental dyslexia. However, the role of phonological awareness in the acquisition of reading Japanese kana, the transparent, mora-based phonogram, has not been understood completely. We examine the abilities of Japanese dyslexic children on different types of Japanese phonological tasks, and discussed which tasks significantly account for each aspect of reading ability. METHODS: Fifteen dyslexic children (9.53+/-1.52 years old), and 15 children with normal reading ability (9.17+/-0.90 years old) participated in this study. They performed three types of phonological awareness tasks. The mora counting task and the mora reversal task of words require phonological awareness at the mora level. The letter rhyming task, which resembles the task in English language, requires phonological awareness at the phoneme level. We evaluated the reading ability by the reading speed, the reading errors, and the number of pauses while reading sentences aloud. RESULTS: The task performances of the dyslexic group on all three phonological awareness tasks were significantly lower than those of the control group. Stepwise multiple regression analysis revealed that the mora counting task and the rhyming letter task most significantly explained the reading speed and number of reading pauses. The mora reversal task of words, together with the antegraded digit span, significantly explained the reading errors. CONCLUSIONS: Japanese dyslexics showed deficits of phonological awareness both at the mora and the phoneme levels. Phonological awareness must be crucial for acquiring the ability of decoding phonograms, including Japanese kana.

Evaluation of Trace Elements in Augmentation of Statin-Induced Cytotoxicity in Uremic Serum-Exposed Human Rhabdomyosarcoma Cells.

Patients with end-stage kidney disease (ESKD) are at higher risk for rhabdomyolysis induced by statin than patients with normal kidney function. Previously, we showed that this increase in the severity of statin-induced rhabdomyolysis was partly due to uremic toxins. However, changes in the quantity of various trace elements in ESKD patients likely contribute as well. The purpose of this study is to determine the effect of trace elements on statin-induced toxicity in rhabdomyosarcoma cells exposed to uremic serum (US cells) for a long time. Cell viability, apoptosis, mRNA expression, and intracellular trace elements were assessed by viability assays, flow cytometry, real-time RT-PCR, and ICP-MS, respectively. US cells exhibited greater simvastatin-induced cytotoxicity than cells long-time exposed with normal serum (NS cells) (non-overlapping 95% confidence intervals). Intracellular levels of Mg, Mn, Cu, and Zn were significantly less in US cells compared to that in NS cells (p < 0.05 or 0.01). Pre-treatment with TPEN increased simvastatin-induced cytotoxicity and eliminated the distinction between both cells of simvastatin-induced cytotoxicity. These results suggest that Zn deficiencies may be involved in the increased risk for muscle complaints in ESKD patients. In conclusion, the increased severity of statin-induced rhabdomyolysis in ESKD patients may be partly due to trace elements deficiencies.

Neural substrates of sarcasm: a functional magnetic-resonance imaging study.

The understanding of sarcasm reflects a complex process, which involves recognizing the beliefs of the speaker. There is a clear association between deficits in mentalizing, which is the ability to understand other people's behavior in terms of their mental state, and the understanding of sarcasm in individuals with autistic spectrum disorders. This suggests that mentalizing is important in pragmatic non-literal language comprehension. To highlight the neural substrates of sarcasm, 20 normal adult volunteers underwent functional magnetic-resonance imaging. We used scenario-reading tasks, in which sentences describing a certain situation were presented, followed by the protagonist's comments regarding that situation. Depending on the situation, the semantic content of the comments was classified as sarcastic, non-sarcastic, or contextually unconnected. As the combination of the first and second sentences represented discourse-level information that was not encoded in the individual sentences, sarcasm detection was represented as the differential activation induced by the second sentences. Sarcasm detection activated the left temporal pole, the superior temporal sulcus, the medial prefrontal cortex, and the inferior frontal gyrus (Brodmann's area [BA] 47). The left BA 47 was activated more prominently by sarcasm detection than by the first sentence. These findings indicate that the detection of sarcasm recruits the medial prefrontal cortex, which is part of the mentalizing system, as well as the neural substrates involved in reading sentences. The left BA 47 might therefore be where mentalizing and language processes interact during sarcasm detection.