Genetic and functional diversity of ß-N-acetylgalactosamine-targeting glycosidases expanded by deep-sea metagenome analysis.

ß-N-Acetylgalactosamine-containing glycans play essential roles in several biological processes, including cell adhesion, signal transduction, and immune responses. ß-N-Acetylgalactosaminidases hydrolyze ß-N-acetylgalactosamine linkages of various glycoconjugates. However, their biological significance remains ambiguous, primarily because only one type of enzyme, exo-ß-N-acetylgalactosaminidases that specifically act on ß-N-acetylgalactosamine residues, has been documented to date. In this study, we identify four groups distributed among all three domains of life and characterize eight ß-N-acetylgalactosaminidases and ß-N-acetylhexosaminidase through sequence-based screening of deep-sea metagenomes and subsequent searching of public protein databases. Despite low sequence similarity, the crystal structures of these enzymes demonstrate that all enzymes share a prototype structure and have diversified their substrate specificities (oligosaccharide-releasing, oligosaccharide/monosaccharide-releasing, and monosaccharide-releasing) through the accumulation of mutations and insertional amino acid sequences. The diverse ß-N-acetylgalactosaminidases reported in this study could facilitate the comprehension of their structures and functions and present evolutionary pathways for expanding their substrate specificity.

Transthyretin Variant Amyloidosis with a TTR A97D (p.A117D) Mutation Manifesting Remarkable Asymmetric Neuropathy.

We herein report a 68-year-old Japanese man with sporadic variant transthyretin (ATTRv) amyloidosis harboring the novel variant A97D (p.A117D) in TTR. He had slow development of asymmetric neuropathy, unintentional weight loss, mild autonomic failure and mild cardiomyopathy. TTR amyloid deposition on the gastric duodenal mucosa was detected. In silico analyses predicted that TTR A97D (p.A117D) altered the structure and function of the TTR protein. ATTRv amyloidosis is often difficult to diagnose in non-endemic regions due to its diverse phenotypes, such as atypical peripheral nerve involvement and a rare family history.

Retained absolute pitch after selective amygdalohippocampectomy.

This study assessed the pre-operative chronic condition and effect of epilepsy surgery in a 21-year-old Japanese woman with drug-resistant right temporal lobe epilepsy (TLE). For this patient, it was crucially important to preserve language and her music capabilities, including absolute pitch (AP), which is found in the general population at less than 0.1%. The patient became seizure free, and her AP capability was preserved after selective amygdalohippocampectomy in the non-dominant right hemisphere. Most of the neuropsychological test (WAIS-III and WMS-R) scores remained in the normal range, except for low scores in verbal memory and markedly improved attention/concentration index. The patient's pre- and postoperative brain function related to language and music capabilities were investigated using functional magnetic resonance imaging (fMRI) based on two language tasks and a music task (listening to melodies). While task performance was similar in pre- and postoperative examinations, her brain activation patterns markedly differed. The most striking difference was during the music task: areas with significant activation existed in the bilateral frontal and temporal lobes before surgery, whereas postoperative activation was confined to a very limited region in the left angular gyrus. The authors speculate that the surgery triggered some change in functional organization in the brain, which contributed to preserving her capabilities.

Analyzing Neural Activity and Connectivity Using Intracranial EEG Data with SPM Software.

Measuring neural activity and connectivity associated with cognitive functions at high spatial and temporal resolutions is an important goal in cognitive neuroscience. Intracranial electroencephalography (EEG) can directly record electrical neural activity and has the unique potential to accomplish this goal. Traditionally, averaging analysis has been applied to analyze intracranial EEG data; however, several new techniques are available for depicting neural activity and intra- and inter-regional connectivity. Here, we introduce two analytical protocols we recently applied to analyze intracranial EEG data using the Statistical Parametric Mapping (SPM) software: time-frequency SPM analysis for neural activity and dynamic causal modeling of induced responses for intra- and inter-regional connectivity. We report our analysis of intracranial EEG data during the observation of faces as representative results. The results revealed that the inferior occipital gyrus (IOG) showed gamma-band activity at very early stages (110 ms) in response to faces, and both the IOG and amygdala showed rapid intra- and inter-regional connectivity using various types of oscillations. These analytical protocols have the potential to identify the neural mechanisms underlying cognitive functions with high spatial and temporal profiles.

Working memory deficit in drug-resistant epilepsy with an amygdala lesion.

This study compared temporal lobe epilepsy (TLE) patients with amygdala lesion (AL) without hippocampal sclerosis (HS) (TLE-AL) with patients with TLE and HS without AL (TLE-HS). Both subtypes of TLE arose from the right hemisphere. The TLE-AL group exhibited a lower Working Memory Index (WMI) on the Wechsler Adult Intelligence Scale, third edition (WAIS-III), indicating that the amygdala in the right hemisphere is involved in memory-related function. [18F]fluorodeoxyglucose positron emission topography (FDG-PET) showed glucose hypometabolism limited to the right uncus for the TLE-AL group. The results suggest the importance of considering cognitive functions in the non-dominant hemisphere to prevent impairment after surgery.

Bidirectional electric communication between the inferior occipital gyrus and the amygdala during face processing.

Faces contain multifaceted information that is important for human communication. Neuroimaging studies have revealed face-specific activation in multiple brain regions, including the inferior occipital gyrus (IOG) and amygdala; it is often assumed that these regions constitute the neural network responsible for the processing of faces. However, it remains unknown whether and how these brain regions transmit information during face processing. This study investigated these questions by applying dynamic causal modeling of induced responses to human intracranial electroencephalography data recorded from the IOG and amygdala during the observation of faces, mosaics, and houses in upright and inverted orientations. Model comparisons assessing the experimental effects of upright faces versus upright houses and upright faces versus upright mosaics consistently indicated that the model having face-specific bidirectional modulatory effects between the IOG and amygdala was the most probable. The experimental effect between upright versus inverted faces also favored the model with bidirectional modulatory effects between the IOG and amygdala. The spectral profiles of modulatory effects revealed both same-frequency (e.g., gamma-gamma) and cross-frequency (e.g., theta-gamma) couplings. These results suggest that the IOG and amygdala communicate rapidly with each other using various types of oscillations for the efficient processing of faces. Hum Brain Mapp 38:4511-4524, 2017. © 2017 Wiley Periodicals, Inc.

Characteristics of EEG Seizure-Onset Patterns Recorded From Subdural Electrodes Over MRI-Visible Frontal Focal Cortical Dysplasia Type IIb Lesions.

PURPOSE: Focal cortical dysplasia (FCD) is intrinsically epileptogenic, and an MRI-visible lesion typically constitutes the core part of the epileptogenic zone. We aimed to identify ictal EEG patterns that represent the epileptogenic zone by using subdural electrodes placed over the MRI-visible FCD lesion. METHODS: We selected seven patients with frontal lobe epilepsy caused by pathologically proven FCD type IIb who underwent preoperative intracranial EEG evaluation with subdural electrodes followed by resection surgery with seizure-free outcome. The characteristics of ictal EEG patterns, interictal/ictal high-frequency oscillations, and ictal direct current shifts from intracranial electrodes placed over the MRI-visible lesion were analyzed. RESULTS: Two seizure-onset patterns (low voltage fast activity and fast spike activity) were identified. Low voltage fast activity was seen in all patients with a lateral frontal lesion, and it was always preceded by preictal spikes. Fast spike activity occurred only in patients with a mesial frontal lesion. Interictal/ictal high-frequency oscillations and ictal direct current shifts were seen in all patients. CONCLUSIONS: The epileptogenic zone of frontal FCD type IIb may be characterized by EEG seizure-onset patterns consisting of low voltage fast activity and fast spike activity accompanied by ictal high-frequency oscillations and ictal direct current shifts. Further study is needed to determine whether other seizure-onset patterns exist in patients with FCD type IIb.

Rapid gamma oscillations in the inferior occipital gyrus in response to eyes.

Eyes are an indispensable communication medium for human social interactions. Although previous neuroscientific evidence suggests the activation of the inferior occipital gyrus (IOG) during eye processing, the temporal profile of this activation remains unclear. To investigate this issue, we analyzed intracranial electroencephalograms of the IOG during the presentation of eyes and mosaics, in either averted or straight directions. Time-frequency statistical parametric mapping analyses revealed greater gamma-band activation in the right IOG beginning at 114 ms in response to eyes relative to mosaics, irrespective of their averted or straight direction. These results suggest that gamma oscillations in the right IOG are involved in the early stages of eye processing, such as eye detection.

Gamma Oscillations in the Temporal Pole in Response to Eyes.

The eyes of an individual act as an indispensable communication medium during human social interactions. Functional neuroimaging studies have revealed that several brain regions are activated in response to eyes and eye gaze direction changes. However, it remains unclear whether the temporal pole is one of these regions. Furthermore, if the temporal pole is activated by these stimuli, the timing and manner in which it is activated also remain unclear. To investigate these issues, we analyzed intracranial electroencephalographic data from the temporal pole that were obtained during the presentation of eyes and mosaics in averted or straight directions and their directional changes. Time-frequency statistical parametric mapping analyses revealed that the bilateral temporal poles exhibited greater gamma-band activation beginning at 215 ms in response to eyes compared with mosaics, irrespective of the direction. Additionally, the right temporal pole showed greater gamma-band activation beginning at 197 ms in response to directional changes of the eyes compared with mosaics. These results suggest that gamma-band oscillations in the temporal pole were involved in the processing of the presence of eyes and changes in eye gaze direction at a relatively late temporal stage compared with the posterior cortices.

Etiology of Sudden Cardiac Arrest in Patients with Epilepsy: Experience of Tertiary Referral Hospital in Sapporo City, Japan.

It has been reported that epilepsy patients had higher risk of sudden death than that of the general population. However, in Japan, there is very little literature on the observational research conducted on sudden fatal events in epilepsy. We performed a single-center, retrospective study on all the out-of-hospital cardiac arrest (OHCA) patients treated in our emergency department between 2007 and 2013. Among the OHCA patients, we extracted those with a history of epilepsy and then analyzed the characteristics of the fatal events and the background of epilepsy. From 1,823 OHCA patients, a total of 10 cases were enrolled in our study. The median age was 34 years at the time of the incident [9-52 years; interquartile range (IQR), 24-45]. We determined that half of our cases resulted from external causes of death such as drowning and suffocation and the other half were classified as sudden unexpected death in epilepsy (SUDEP). In addition, asphyxia was implicated as the cause in eight cases. Only the two near-drowning patients were immediately resuscitated, but the remaining eight patients died. The median age of first onset of epilepsy was 12 years (0.5-30; IQR, 3-21), and the median disease duration was 25 years (4-38; IQR, 6-32). Patients with active epilepsy accounted for half of our series and they were undergoing poly anti-epileptic drug therapy. The fatal events related to epilepsy tended to occur in the younger adult by external causes. An appropriate therapeutic intervention and a thorough observation were needed for its prevention.

Involvement of flocculin in negative potential-applied ITO electrode adhesion of yeast cells.

The purpose of this study was to develop novel methods for attachment and cultivation of specifically positioned single yeast cells on a microelectrode surface with the application of a weak electrical potential. Saccharomyces cerevisiae diploid strains attached to an indium tin oxide/glass (ITO) electrode to which a negative potential between -0.2 and -0.4 V vs. Ag/AgCl was applied, while they did not adhere to a gallium-doped zinc oxide/glass electrode surface. The yeast cells attached to the negative potential-applied ITO electrodes showed normal cell proliferation. We found that the flocculin FLO10 gene-disrupted diploid BY4743 mutant strain (flo10Δ /flo10Δ) almost completely lost the ability to adhere to the negative potential-applied ITO electrode. Our results indicate that the mechanisms of diploid BY4743 S. cerevisiae adhesion involve interaction between the negative potential-applied ITO electrode and the Flo10 protein on the cell wall surface. A combination of micropatterning techniques of living single yeast cell on the ITO electrode and omics technologies holds potential of novel, highly parallelized, microchip-based single-cell analysis that will contribute to new screening concepts and applications.

Significance of Very-High-Frequency Oscillations (Over 1,000Hz) in Epilepsy.

OBJECTIVE: We previously reported ictal very-high-frequency oscillations (VHFO) of 1,000 to 2,500Hz recorded by subdural macroelectrodes using a 10-kHz sampling rate. The purpose of this study was to clarify the clinical significance of ictal VHFO in neocortical epilepsy. METHODS: This study included 13 patients with neocortical epilepsy who underwent subdural electrode implantation and had at least 1 seizure recorded at a 10-kHz sampling rate and were followed for more than 2 years postoperatively. Extent of resection was determined considering the seizure onset zone (SOZ) and irritative zone, structural lesion, and functional areas. Areas showing VHFO and those with HFO were not taken into consideration. The presence or absence of VHFO (>1,000 Hz), HFO (200-1,000Hz) and SOZ, and completeness of resection of these areas were compared with postoperative seizure outcome. RESULTS: Seven patients had favorable (Engel class Ia) and 6 had unfavorable outcomes (other classes). VHFO was recorded in 6 of 7 patients with a favorable outcome. On the contrary, VHFO was recorded in only 1 of 6 patients with unfavorable outcome. The presence of VHFO was significantly associated with favorable outcome. VHFO was recorded on a limited number of electrodes, and VHFO-generating areas were resected completely, whereas HFO-generating areas and/or SOZ were not always resected completely in both favorable and unfavorable outcome groups. INTERPRETATION: The presence of ictal VHFO may be predictive of favorable outcome. Ictal VHFO may be a more specific marker than ictal HFO or SOZ for identifying the core of epileptogenic zone.

Identification of HSP70-inducing activity in Arnica montana extract and purification and characterization of HSP70-inducers.

BACKGROUND: The expression of heat shock proteins (HSPs), particularly HSP70, is receiving considerable attention in the field of cosmetics, particularly given our recent report that ultraviolet-induced melanin production, skin damage and wrinkle formation were all suppressed in transgenic mice expressing HSP70. OBJECTIVE: In the present study, we searched for HSP70-inducers from a library of herbal extracts that have already been approved as quasi-pharmaceutical products in Japan. We selected an ethanol extract of Arnica montana (A. montana), based on its high HSP70-inducing activity and low cytotoxicity. METHODS: Cell viability was determined by MTT method and expression of HPS70 was monitored by immunoblotting analysis. RESULTS: From the extract, we purified and identified eight sesquiterpene lactones (AM1-8) as HSP70-inducers, among which AM-2 (helenalin 2-methylbutyrate) was selected due to its good HSP70-inducing properties and low cytotoxicity. Treatment of cultured mouse melanoma cells with AM-2 or A. montana extract up-regulated the expression of HSP70 in a dose-dependent manner. This treatment also activated heat shock factor-1, a transcription factor for hsp genes. Furthermore, pre-treatment of cells with AM-2 or A. montana extract decreased melanin production and expression and activity of tyrosinase. CONCLUSION: These results suggest that AM-2 and A. montana extract could be beneficial for use in hypopigmenting cosmetics as a consequence of their stimulatory effects on HSP70 expression.

Aneurinibacillus tyrosinisolvens sp. nov., a tyrosine-dissolving bacterium isolated from organics- and methane-rich seafloor sediment.

A novel Gram-positive-staining, strictly aerobic and heterotrophic bacterium, designated strain LL-002T, was isolated from organics- and methane-rich seafloor sediment at a depth of 100 m in Kagoshima Bay, Kagoshima, Japan. Colonies were lustreless and translucent white in colour. The temperature, pH and salt concentration ranges for growth were 10-30 °C, pH 6.0-6.5 and 0-1 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that strain LL-002T belongs to the genus Aneurinibacillus of the family Paenibacillaceae. 16S rRNA gene sequence similarities between strain LL-002T and the type strains of species of the genus Aneurinibacillus were 92.8-95.7 %; the highest sequence identity was with the type strain of Aneurinibacillus migulanus. The DNA G+C content of strain LL-002T was 46.2 mol%. MK-7 was the predominant menaquinone. The predominant cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0, and the cell-wall peptidoglycan contained meso-diaminopimelic acid and glutamic acid, glycine and alanine in addition to muramic acid and glucosamine. The peptidoglycan type was A1γ. In DNA-DNA hybridization assays between strain LL-002T and the type strains of the other species of the genus Aneurinibacillus, the level of hybridization was 6.3-30.1 %. On the basis of its biological features and the 16S rRNA gene sequence comparison presented here, strain LL-002T is considered to represent a novel species of the genus Aneurinibacillus, for which the name Aneurinibacillus tyrosinisolvens sp. nov. is proposed; the type strain is LL-002T ( = NBRC 110097T = CECT 8536T).

Halarchaeum grantii sp. nov., a moderately acidophilic haloarchaeon isolated from a commercial salt sample.

Three moderately acidophilic, halophilic archaeal strains, MH1-243-3T, MH1-243-5 and MH1-243-6, were isolated from a commercial salt sample made from seawater in Okinawa, Japan. Cells of the three strains were pleomorphic and stained Gram-negative. Colonies of the strains were orange-red-pigmented. Strain MH1-243-3T was able to grow at 15-27 % (w/v) NaCl (optimum 24 °C), at pH 4.5-6.5 (pH 5.5) and at 35-50 °C (45 °C). Strains MH1-243-5 and MH1-243-6 grew within slightly different ranges (shown in text). The 16S rRNA gene sequences of the three strains were identical, and the closest phylogenetic relative was Halarchaeum salinum MH1-34-1T with 97.0 % similarity. The rpoB' gene sequences of the three strains were also identical, and the closest phylogenetic relative was Halarchaeum acidiphilum JCM 16109T with 92.0 % similarity. The DNA G+C content of MH1-243-3T, MH1-243-5 and MH1-243-6 was 65.2 mol%. The levels of DNA-DNA relatedness amongst the three strains were 84.1-99.8 %, while that between MH1-243-3T and H. salinum MH1-34-1T was 30.6 % and 31.6 % (reciprocally), and those between MH1-243-3T and type strains of other species in the genus Halarchaeum were 42.3-29.4 %. Based on the phenotypic, genotypic and phylogenetic analyses, it is proposed that the isolates should represent a novel species of the genus Halarchaeum, for which the name Halarchaeum grantii sp. nov. is proposed. The type strain is MH1-243-3T ( = JCM 19585T = KCTC 4142T), isolated from commercial sea salt produced in Okinawa, Japan. MH1-243-5 ( = JCM 19586) and MH1-243-6 ( = JCM 18422) are additional strains of the species.

Bidirectional neural connectivity between basal temporal and posterior language areas in humans.

OBJECTIVE: The basal temporal language area (BTL) is known to be involved in the semantic processing of language. To investigate the neural connectivity between BTL and the posterior language area (PL), we used cortico-cortical evoked potential (CCEP) technique. METHODS: Four patients with intractable epilepsy who underwent presurgical evaluation with subdural electrodes were examined. All patients were right-handed and left language dominance by Wada test. We directly stimulated 20 pairs of electrodes placed on BTL in patient 1-3, putative BTL in patient 4, and PL in patient 1-4. In patient 4, all electrodes on the left temporal basal area were stimulated. RESULTS: We could record 132 CCEP responses including 40 responses by the left basal temporal stimulation in patient 4. The waveforms from PL to BTL were triphasic, while those from BTL to PL were biphasic. The mean latency of the first negative peak (N1) was shorter at BTL (31.8-41.0ms; mean 35.1ms) than at PL (39.6-73.2ms; mean 52.3ms). CONCLUSIONS: We revealed the uneven bidirectional connection between BTL and PL. SIGNIFICANCE: We speculated that the two language areas are connected mainly through subcortical fibers from PL to BTL and through cortico-cortical fibers from BTL to PL, mediated by multisynaptic transmissions.

Preservation of absolute pitch after right amygdalohippocampectomy for a pianist with TLE.

Absolute pitch (AP) ability is a rare musical phenomenon. In the literature, it has been suggested that the relative specialization for pitch processing is in the right temporal lobe in the non-AP population. Since the anatomic basis for absolute pitch is not fully understood and cases of temporal lobe epilepsy of AP possessors are extremely rare, applicability of resection as a treatment of epilepsy in this particular area should be evaluated with caution. In the present study, we examined an AP possessor who suffered from medically refractory temporal lobe epilepsy and underwent right selective amygdalohippocampectomy (SAH). The SAH procedure clearly avoided disturbing important structures for AP, inasmuch as postsurgically she preserved her AP ability and was seizure-free. She did well post-operatively in the test of pure sine wave tones with short reaction time, which could be identified as "true" absolute pitch.

Brevundimonas denitrificans sp. nov., a denitrifying bacterium isolated from deep subseafloor sediment.

A novel Gram-stain-negative, aerobic, heterotrophic, stalked and capsulated bacterium with potential denitrification ability, designated strain TAR-002(T), was isolated from deep seafloor sediment in Japan. Colonies lacked lustre, and were viscous and translucent white. The ranges of temperature, pH and salt concentration for growth were 8-30 °C, pH 6.0-10.0 and 1-3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that strain TAR-002(T) belongs to the genus Brevundimonas of the class Alphaproteobacteria. Levels of similarity between the 16S rRNA gene sequence of strain TAR-002(T) and those of the type strains of species of the genus Brevundimonas were 93.5-98.9%; the most closely related species was Brevundimonas basaltis. In DNA-DNA hybridization assays between strain TAR-002(T) and its phylogenetic neighbours, Brevundimonas lenta DS-18(T), B. basaltis J22(T), Brevundimonas subvibrioides ATCC 15264(T) and Brevundimonas alba DSM 4736(T), mean hybridization levels were 6.4-27.7%. The G+C content of strain TAR-002(T) was 70.3 mol%. Q-10 was the major respiratory isoprenoid quinone. The major fatty acids were C(18:1)ω7c and C(16:0), and the presence of 1,2-di-O-acyl-3-O-[D-glucopyranosyl-(1 → 4)-α-D-glucopyranuronosyl]glycerol (DGL) indicates the affiliation of strain TAR-002(T) with the genus Brevundimonas. On the basis of biological characteristics and 16S rRNA gene sequence comparisons, strain TAR-002(T) is considered to represent a novel species of the genus Brevundimonas, for which the name Brevundimonas denitrificans sp. nov. is proposed; the type strain is TAR-002(T) ( =NBRC 110107(T) =CECT 8537(T)).

Rapid, high-frequency, and theta-coupled gamma oscillations in the inferior occipital gyrus during face processing.

Neuroimaging studies have found greater activation in the inferior occipital gyrus (IOG), or occipital face area, in response to faces relative to non-facial stimuli. However, the temporal, frequency, and functional profiles of IOG activity during face processing remain unclear. Here, this issue was investigated by recording intracranial field potentials in the IOG during the presentation of faces, mosaics, and houses in upright and inverted orientations. Time-frequency statistical parametric mapping analyses revealed greater gamma-band activation in the IOG beginning at 110 msec and covering 40-300 Hz in response to upright faces relative to upright houses and mosaics. Phase-amplitude cross-frequency coupling analyses revealed more evident theta-gamma couplings at 115-256 msec during the processing of upright faces as compared with that of upright houses and mosaics. Comparable gamma-band activity was observed during the processing of inverted and upright faces at about 100-200 msec, but weaker activity and different coupling with theta-band activity after 200 msec. These patterns of activity were more evident in the right than in the left IOG. These results, together with other evidence on neural communication, suggest that broadband gamma oscillations in the right IOG conduct rapid and multistage (i.e., both featural and configural) face processing in collaboration with theta oscillations transmitted from other brain regions.