Increased resistance against tellurite is conferred by a mutation in the promoter region of uncommon tellurite resistance gene tehB in the ter-negative Shiga toxin-producing Escherichia coli O157:H7.

Resistance to potassium tellurite (PT) is an important indicator in isolating Shiga toxin-producing Escherichia coli (STEC) O157:H7 and other major STEC serogroups. Common resistance determinant genes are encoded in the ter gene cluster. We found an O157:H7 isolate that does not harbor ter but is resistant to PT. One nonsynonymous mutation was found in another PT resistance gene, tehA, through whole-genome sequence analyses. To elucidate the contribution of this mutation to PT resistance, complementation of tehA and the related gene tehB in isogenic strains and quantitative RT‒PCR were performed. The results indicated that the point mutation not only changed an amino acid of tehA, but also was positioned on a putative internal promoter of tehB and increased PT resistance by elevating tehB mRNA expression. Meanwhile, the amino acid change in tehA had negligible impact on the PT resistance. Comprehensive screening revealed that 2.3% of O157:H7 isolates in Japan did not harbor the ter gene cluster, but the same mutation in tehA was not found. These results suggested that PT resistance in E. coli can be enhanced through one mutational event even in ter-negative strains. IMPORTANCE: Selective agents are important for isolating Shiga toxin-producing Escherichia coli (STEC) because the undesirable growth of microflora should be inhibited. Potassium tellurite (PT) is a common selective agent for major STEC serotypes. In this study, we found a novel variant of PT resistance genes, tehAB, in STEC O157:H7. Molecular experiments clearly showed that one point mutation in a predicted internal promoter region of tehB upregulated the expression of the gene and consequently led to increased resistance to PT. Because tehAB genes are ubiquitous across E. coli, these results provide universal insight into PT resistance in this species.

Trans men can achieve adequate muscular development through low-dose testosterone therapy: A long-term study on body composition changes.

BACKGROUND: Transgender individuals undergo the gender-affirming hormone therapy (GAHT) to achieve physical changes consistent with their gender identity. Few studies are available on the long-term safety and efficacy of GAHT. OBJECTIVES: To investigate the long-term physical effects and the safety of the testosterone therapy for trans men and to assess the impact of differential hormone dose. MATERIALS AND METHODS: Trans men who initiated GAHT between May 2000 and December 2021 were included in this retrospective analysis. Physical findings (body mass index, body fat percentage (BFP), lean body mass (LBM), and grip strength), blood testing results (hemoglobin, hematocrit, uric acid, creatinine, total cholesterol, triglycerides, and total testosterone), and menstrual cessation were recorded. We assessed the effects of testosterone on body composition changes and laboratory parameters, comparing a low-dose group (≤ 62.5 mg/wk) to a high-dose group (> 62.5 mg/wk). RESULTS: Of 291 participants, 188 patients (64.6%) were in the low-dose group and 103 (35.4%) in the high-dose group. Cumulative menstrual cessation rates up to 12 months were not significantly different between groups. Both groups showed a decrease in BFP and an increase in LBM during the first year of therapy, followed by a slight increase in both over the long term. The high-dose group exhibited greater LBM gains during the first year. Higher hormone doses and lower initial LBM values were associated with LBM increases at 3 and 6 months (3 mo, P = 0.006, P < 0.001; 6 mo, P = 0.015, P < 0.001). There were no long-term, dose-dependent side effects such as polycythemia or dyslipidemia. CONCLUSION: Long-term GAHT for trans men is safe and effective. Low-dose testosterone administration is sufficient to increase LBM in trans men. Higher testosterone doses can lead to an earlier increase in muscle mass.

Alternative autophagy dampens UVB-induced NLRP3 inflammasome activation in human keratinocytes.

Sunlight exposure results in an inflammatory reaction of the skin commonly known as sunburn, which increases skin cancer risk. In particular, the ultraviolet B (UVB) component of sunlight induces inflammasome activation in keratinocytes to instigate the cutaneous inflammatory responses. Here, we explore the intracellular machinery that maintains skin homeostasis by suppressing UVB-induced inflammasome activation in human keratinocytes. We found that pharmacological inhibition of autophagy promoted UVB-induced NLRP3 inflammasome activation. Unexpectedly, however, gene silencing of Atg5 or Atg7, which are critical for conventional autophagy, had no effect, whereas gene silencing of Beclin1, which is essential not only for conventional autophagy but also for Atg5/Atg7-independent alternative autophagy, promoted UVB-induced inflammasome activation, indicating an involvement of alternative autophagy. We found that damaged mitochondria were highly accumulated in UVB-irradiated keratinocytes when alternative autophagy was inhibited, and they appear to be recognized by NLRP3. Overall, our findings indicate that alternative autophagy, rather than conventional autophagy, suppresses UVB-induced NLRP3 inflammasome activation through the clearance of damaged mitochondria in human keratinocytes and illustrate a previously unknown involvement of alternative autophagy in inflammation. Alternative autophagy may be a new therapeutic target for sunburn and associated cutaneous disorders.

A Qualitative Study on the Development of Professionalism Among Japanese Dental Students.

BACKGROUND: Professionalism is believed to vary depending on factors such as era and culture. Therefore, clarifying the meaning of professionalism in each country, region, and workplace is essential. However, how professionalism is cultivated among dental students in Japanese schools has yet to be fully elucidated. Therefore, this study examined whether professionalism among Japanese dental students changes by year. This research will contribute to effective professional education. PARTICIPANTS AND METHODS: The participants included six fourth-year dental students and nine fifth-year dental students. Semi-structured interviews were conducted from November 2018 to January 2019, and verbatim transcripts were created from the recorded data. Based on these verbatim transcripts, thematic analysis was utilized to examine and identify professionalism components for each academic year. RESULTS: Three themes based on 14 constituent concepts were obtained for fourth-year students. Three themes based on 20 constituent concepts were obtained for fifth-year students. Fourth-year students primarily focused on technical aspects. In contrast, fifth-year students placed greater emphasis on attitude and communication skills. CONCLUSION: From fourth-year students, who primarily focus on classroom learning and practical training, to fifth-year students who gain clinical experience, the constituent elements of professionalism became more complex. However, this study did not examine other aspects of healthcare professionalism, such as interprofessional collaboration. A comprehensive education program tailored to the clinical setting is necessary for cultivating professionalism.

Talk to us about your skin: The relationship between spoken language and haptic exploratory procedures.

OBJECTIVE: To enhance satisfaction, the cosmetics industry needs to clearly understand consumers' descriptions of their key tactile preferences. It is difficult for researchers to understand verbal descriptions from people whose native language is different from their own. Previous research has implied that some sensory words with the same lexical meanings have been observed in different haptic exploratory procedures (HEPs). Therefore, our study aims to investigate and understand the key tactile perceptions of people from five different countries based on their descriptions and their HEPs. METHODS: In Experiment 1, 1545 participants living in the US, Japan, China, Italy, and Thailand described their major tactile perceptions as efficacy in skincare, and we analysed the frequency of each word used in their answers. In Experiment 2, we confirmed the task to observe HEPs for Chinese, Italian, and Thai participants. A total of 24 participants in China, 33 participants in Italy, and 30 participants in Thailand freely explored their faces with their hands and answered which side more closely matched the major tactile adjectives. Experts classified the observed HEPs into six classifications within two categories and three contact area sizes and investigated the cultural differences. RESULTS: More than 2% of the Chinese, Italian, Thai, US, and Japanese participants described 33, 20, 29, 22, and 18 words, respectively, as efficacy in skincare. Verified words that described the major tactile perceptions in each native language had the same meanings as moistness, firmness, softness, smoothness, and so on. We could confirm the HEPs of these major feelings for the participants from each culture. Chinese and Thai participants' HEPs for moistness or softness were observed with a pressing movement. Conversely, Italian participants' HEPs for moistness or softness were observed with a rubbing movement. CONCLUSION: This study showed that words with the same lexical meanings evoked different HEPs. The results imply that different HEPs can provide different physical stimuli on the skin. Therefore, it is important to survey both objects and HEPs to better understand the tactile experience.

OBJECTIF: Pour améliorer la satisfaction, l'industrie cosmétique doit bien comprendre les descriptions que font les consommateurs de leurs principales préférences tactiles. Il est difficile pour les chercheurs de comprendre les descriptions verbales des personnes dont la langue maternelle est différente de la leur. Des recherches antérieures ont suggéré que certains mots sensoriels ayant les mêmes significations lexicales ont été observés dans différentes procédures exploratoires haptiques (PEH). Par conséquent, notre étude vise à étudier et à comprendre les perceptions tactiles clés des personnes de cinq pays différents en fonction de leurs descriptions et de leurs PEH. MÉTHODES: Dans l'expérience 1 545 participantes vivant aux États­Unis, au Japon, en Chine, en Italie et en Thaïlande ont décrit leurs principales perceptions tactiles comme l'efficacité dans les soins de la peau, et nous avons analysé la fréquence de chaque mot utilisé dans leurs réponses. Dans l'expérience 2, nous l'avons confirmé en observant les PEH pour les participantes chinoises, italiennes et thaïs. 24 participantes en Chine, 33 participantes en Italie et 30 participantes en Thaïlande ont librement exploré leur visage avec leurs mains et ont répondu à la question de savoir quel côté correspondait le mieux aux principaux adjectifs tactiles. Les experts ont classé les PEH observées en six classifications dans deux catégories et trois tailles de surface de contact, et ont étudié les différences culturelles. RÉSULTAT: Plus de deux pour cent des participantes chinoises, italiennes, thaïs, américaines et japonaises ont décrit 33, 20, 29, 22 et 18 mots, respectivement, comme une efficacité dans les soins de la peau. Les mots vérifiés qui décrivaient les principales perceptions tactiles dans chaque langue maternelle ayant les mêmes significations sont l'humidité, la fermeté, la douceur, la texture lisse, etc. Nous avons pu confirmer les PEH de ces sensations majeures pour les participants de chaque culture. Les PEH des participantes chinoises et thaïs pour l'humidité ou la douceur ont été observées avec un mouvement de pression. A l'inverse, les PEH pour l'humidité ou la douceur des participantes italiennes ont été observées avec un mouvement de frottement. CONCLUSION: Cette étude a montré que les mots ayant les mêmes significations lexicales évoquaient différentes PEH. Les résultats impliquent que différentes PEH peuvent fournir différents stimuli physiques sur la peau. Par conséquent, il est important d'étudier les objets et les PEH pour mieux comprendre l'expérience tactile.

The effects of chest drainage on pressure-controlled ventilation.

BACKGROUND: The use of pressure-controlled ventilation (PCV) for anesthesia management is becoming more commonly used. Chest drainage is commonly performed after thoracic surgery, and the negative pressure it generates might affect the transpulmonary pressure (TPP). In the present study, we investigated how chest drainage could affect ventilating conditions during PCV. METHODS: We created a hand-made simple thoracic and lung model, which was connected to an anesthesia machine. The tidal volume (TV) was measured with positive end-expiratory pressure (PEEP) 0 and no chest drainage (baseline), followed by 10 cmH2O PEEP/no drainage, 10 cmH2O PEEP/drainage with - 10 cmH2O and 10 cmH2O PEEP/drainage with - 20 cmH2O. Finally, TV with 20 cmH2O and 30 cmH2O PEEP/no drainage was measured. Driving (inspiratory) pressure was maintained at 20 cmH2O during the whole experiment. RESULTS: TV was significantly increased by applying 10 cmH2O PEEP compared with baseline, further increased by applying - 10 cmH2O by drainage, similar to the value with PEEP 20 cmH2O with no drainage (end-tidal TPP of 20 cmH2O for both). TV decreased to < 50% of the baseline by applying 10 cmH2O PEEP with - 20 cmH2O by drainage, which was similar to that with 30 cmH2O PEEP with no drainage (end-tidal TPP of 30 cmH2O for both). CONCLUSIONS: TV was maintained at similar levels with the same TPP, regardless of PEEP or negative pressure by chest drainage change, suggesting that negative intrapleural pressure by the chest tube drainage system might mimic PEEP from the point of TV.

Evaluation of a Capillary Microsampling Device for Analyzing Plasma Lenvatinib Concentration in Patients With Hepatocellular Carcinoma.

BACKGROUND: The anticancer drug, Lenvima (lenvatinib), has severe side effects. Therapeutic drug monitoring helps ensure its efficacy and safety. Regular and optimally timed blood sampling is tough, especially when lenvatinib is self-medicated. Microsampling using the easy to handle Microsampling Wing (MSW) may help circumvent this problem. However, current lenvatinib detection methods are not sensitive enough to detect its concentrations in microsamples (<50-250 µL). Thus, the aim of this study was 2-fold (1) develop an analytic method to estimate plasma lenvatinib concentrations in microsamples and (2) verify whether this method works on micro (5.6 µL) blood plasma samples obtained clinically through MSW from patients with unresectable hepatocellular carcinoma (HCC). METHODS: A simple, highly sensitive, and specific liquid chromatography-electrospray ionization tandem mass spectrometry method was developed. Using this novel protocol, the trough blood plasma concentration of lenvatinib was measured for both blood sampled conventionally and that using MSW. Thirty-five venous whole blood samples were obtained from 11 patients with HCC. Furthermore, the stability of lenvatinib in MSW samples during storage was evaluated. RESULTS: The mean plasma lenvatinib concentration estimates were not significantly different between the MSW and conventional venous blood samples. CV for interday and intraday assays was low. Up to day 5, the lenvatinib concentration in the MSW samples was 85%-115% of the initial day concentration (when stored at 25°C or 4°C). The interference of endogenous matrix components in the human plasma was low. CONCLUSIONS: These results indicate that the novel mass spectrometry protocol accurately measures lenvatinib in human plasma and is reproducible. Thus, MSW could be a useful microsampling device for lenvatinib therapeutic drug monitoring in patients with HCC when used in combination with this novel liquid chromatography-electrospray ionization tandem mass spectrometry detection method.

The global population structure and evolutionary history of the acquisition of major virulence factor-encoding genetic elements in Shiga toxin-producing Escherichia coli O121:H19.

Shiga toxin (Stx)-producing Escherichia coli (STEC) are foodborne pathogens causing serious diseases, such as haemorrhagic colitis and haemolytic uraemic syndrome. Although O157:H7 STEC strains have been the most prevalent, incidences of STEC infections by several other serotypes have recently increased. O121:H19 STEC is one of these major non-O157 STECs, but systematic whole genome sequence (WGS) analyses have not yet been conducted on this STEC. Here, we performed a global WGS analysis of 638 O121:H19 strains, including 143 sequenced in this study, and a detailed comparison of 11 complete genomes, including four obtained in this study. By serotype-wide WGS analysis, we found that O121:H19 strains were divided into four lineages, including major and second major lineages (named L1 and L3, respectively), and that the locus of enterocyte effacement (LEE) encoding a type III secretion system (T3SS) was acquired by the common ancestor of O121:H19. Analyses of 11 complete genomes belonging to L1 or L3 revealed remarkable interlineage differences in the prophage pool and prophage-encoded T3SS effector repertoire, independent acquisition of virulence plasmids by the two lineages, and high conservation in the prophage repertoire, including that for Stx2a phages in lineage L1. Further sequence determination of complete Stx2a phage genomes of 49 strains confirmed that Stx2a phages in lineage L1 are highly conserved short-tailed phages, while those in lineage L3 are long-tailed lambda-like phages with notable genomic diversity, suggesting that an Stx2a phage was acquired by the common ancestor of L1 and has been stably maintained. Consistent with these genomic features of Stx2a phages, most lineage L1 strains produced much higher levels of Stx2a than lineage L3 strains. Altogether, this study provides a global phylogenetic overview of O121:H19 STEC and shows the interlineage genomic differences and the highly conserved genomic features of the major lineage within this serotype of STEC.

Whole Genome Analysis Detects the Emergence of a Single Salmonella enterica Serovar Chester Clone in Japan's Kanto Region.

In Japan's Kanto region, the number of Salmonella enterica serovar Chester infections increased temporarily between 2014 and 2016. Concurrently with this temporal increase in the Kanto region, S. Chester isolates belonging to one clonal group were causing repetitive outbreaks in Europe. A recent study reported that the European outbreaks were associated with travelers who had been exposed to contaminated food in Morocco, possibly seafood. Because Japan imports a large amount of seafood from Morocco, we aimed to establish whether the temporal increase in S. Chester infections in the Kanto region was associated with imported Moroccan seafood. Short sequence reads from the whole-genome sequencing of 47 S. Chester isolates from people in the Kanto region (2014-2016), and the additional genome sequences from 58 isolates from the European outbreaks, were analyzed. The reads were compared with the complete genome sequence from a S. Chester reference strain, and 347 single nucleotide polymorphisms (SNPs) were identified. These SNPs were used in this study. Cluster and Bayesian cluster analyses showed that the Japanese and European isolates fell into two different clusters. Therefore, Φ PT and I A S values were calculated to evaluate genetic differences between these clusters. The results revealed that the Japanese and European isolates were genetically distinct populations. Our root-to-tip analysis showed that the Japanese isolates originating from one clone had accumulated mutations, suggesting that an emergence of this organism occurred. A minimum spanning tree analysis demonstrated no correlation between genetic and geographical distances in the Japanese isolates, suggesting that the emergence of the serovar in the Kanto region did not involve person-to-person contact; rather, it occurred through food consumption. The d N /d S ratio indicated that the Japanese strain has evolved under positive selection pressure. Generally, a population of bacterial clones in a reservoir faces negative selection pressure. Therefore, the Japanese strain must have existed outside of any reservoir during its emergence. In conclusion, S. Chester isolates originating from one clone probably emerged in the Kanto region via the consumption of contaminated foods other than imported Moroccan seafood. The emerging strain may have not established a reservoir for survival in the food supply chain resulting in its disappearance after 2017.

Subtype Screening of blaIMP Genes Using Bipartite Primers for DNA Sequencing.

Genes conferring carbapenem resistance have spread worldwide among gram-negative bacteria. Subtyping of these genes has epidemiological value due to the global cross-border movement of people. Subtyping of blaIMP genes that frequently detected in Japan appears to be important in public health settings; however, there are few useful tools for this purpose. We developed a subtyping screening tool based on PCR direct sequencing, which targets the internal sequences of almost all blaIMP genes. The tool used bipartite multiplex primers with M13 universal sequences at the 5'-end. According to in silico analysis, among the 78 known IMP-type genes, except for blaIMP-81, 77 detected genes were estimated to be differentiated. In vitro evaluation indicated that sequences of amplicons of IMP-1, IMP-6, IMP-7, and IMP-20 templates were identical to their respective subtypes. Even if the amplicons were small or undetectable through the first PCR, sufficient amplicons for DNA sequencing were obtained through a second PCR using the M13 universal primers. In conclusion, our tool can be possibly used for subtype screening of blaIMP, which is useful for the surveillance of bacteria with blaIMP in clinical and public health settings or environmental fields.

Development of a simple and cost-effective gel-based duplex PCR method to identify both encapsulated and unencapsulated Neisseria meningitidis applicable under resource-limited conditions.

Tens of thousands of cases of invasive meningococcal diseases (IMD) with thousands of deaths are reported annually worldwide; however, only approximately 40 cases occur each year in Japan. Therefore, the majority of medical technologists in Japan have never performed or prepared for analyses of the causative agent, Neisseria meningitidis. Since IMD outbreaks have been reported at mass gathering events, the risk of IMD will increase in Japan in 2021 because of the Olympics. In the present study, we developed a new simple gel-based duplex PCR method that may be employed by the majority Japanese clinical laboratories. It is simple to perform and time- and cost-effectively identifies encapsulated and unencapsulated N. meningitidis by detecting the encapsulated N. meningitidis-specific ctrB and N. meningitidis-specific ggt genes. We consider this simple and cost-effective identification method to compensate for the lack of experience and resource-poor conditions in most Japanese laboratories in which N. meningitidis has rarely been examined.

Inflammasome Sensor NLRP1 Confers Acquired Drug Resistance to Temozolomide in Human Melanoma.

Cancer cells gain drug resistance through a complex mechanism, in which nuclear factor-κB (NF-κB) and interleukin-1ß (IL-1ß) are critical contributors. Because NACHT, LRR and PYD domains-containing protein (NLRP) inflammasomes mediate IL-1ß maturation and NF-κB activation, we investigated the role of inflammasome sensor NLRP1 in acquired drug resistance to temozolomide (TMZ) in melanoma. The sensitivity of melanoma cells to TMZ was negatively correlated with the expression levels of O6-methylguanine-DNA methyltransferase (MGMT), the enzyme to repair TMZ-induced DNA lesions. When MGMT-low human melanoma cells (1205Lu and HS294T) were treated with TMZ for over two months, MGMT was upregulated, and cells became resistant. However, the resistance mechanism was independent of MGMT, and the cells that acquired TMZ resistance showed increased NLRP1 expression, NLRP inflammasome activation, IL-1ß secretion, and NF-κB activity, which contributed to the acquired resistance to TMZ. Finally, blocking IL-1 receptor (IL-1R) signaling with IL-1R antagonist decreased TMZ-resistant 1205Lu tumor growth in vivo. Although inflammation has been associated with drug resistance in various cancers, our paper is the first to demonstrate the involvement of NLRP in the development of acquired drug resistance. Because drug-tolerant cancer cells become cross-tolerant to other classes of cancer drugs, NLRP1 might be a suitable therapeutic target in drug-resistant melanoma, as well as in other cancers.

Cerebral cortex and autonomic nervous system responses during emotional memory processing.

Psychiatric symptoms are often accompanied by somatic symptoms induced by the activity of the autonomic nervous system (ANS). The aim of this study was to calculate the time lag between electroencephalography (EEG) and electrocardiography (ECG) responses, to clarify the changes in the relationship between the cerebral cortex (CC) and the sympathetic nervous system (SNS) during emotional recall processing. Twenty-two healthy young adults were examined. Their EEG and ECG data were simultaneously recorded during emotional audiovisual recall tasks using pleasant and unpleasant stimuli for 180 s, with three repetitions (Epochs 1 & 2 and Epoch 3). The EEG data were analyzed using a fast Fourier transform (FFT) to obtain a time series of relative power spectra, XE, in the theta 1, theta 2, alpha 1, alpha 2, alpha 3, beta 1, beta 2, and beta 3 bands. Time series of RR (inter-beat) intervals (time intervals between successive R waves) derived from the ECG spectral analysis using FFT was applied to the resampled time series of RR intervals over about 60 s to obtain a time series of power spectra for the ratio low frequency/high frequency (LH/HF), XC, which reflects the activity of the sympathetic nervous function. The time lag between XE and XC was calculated using wavelet-crosscorrelation analysis. The results demonstrated that the brain responded to unfamiliar emotionally pleasant stimuli in Epochs 1 & 2 earlier than the SNS, whereas the brain and SNS responded to unfamiliar unpleasant stimuli nearly simultaneously. The brain was activated rapidly in response to familiar unpleasant stimuli, although SNS responded more rapidly to familiar pleasant stimuli than the brain in Epoch 3. Our results quantitatively describe the relationship between the CC and the ANS during emotional recall.

Single-Tube Multiplex Polymerase Chain Reaction for the Detection of Genes Encoding Enterobacteriaceae Carbapenemase.

A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: blaKPC, blaIMP, blaNDM, blaVIM, blaOXA-48-like, and blaGES. Of 70 blaIMP variants, 67 subtypes were simulated to be PCR-positive based on in silico simulation and the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

Attention, Not Performance, Correlates With Afterdischarge Termination During Cortical Stimulation.

Cortical stimulation has been used for brain mapping for over a century, and a standard assumption is that stimulation interferes with task execution due to local effects at the stimulation site. Stimulation can however produce afterdischarges which interfere with functional localization and can lead to unwanted seizures. We previously showed that (a) cognitive effort can terminate these afterdischarges, (b) when termination thus occurs, there are electrocorticography changes throughout the cortex, not just at sites with afterdischarges or sites thought functionally important for the cognitive task used, and (c) thresholds for afterdischarges and functional responses can change among stimulation trials. We here show that afterdischarge termination can occur prior to overt performance of the cognitive tasks used to terminate them. These findings, taken together, demonstrate that task-related brain changes are not limited to one or a group of functional regions or a specific network, and not limited to the time directly surrounding overt task execution. Discrete locations, networks and times importantly underpin clinical behaviors. However, brain activity that is diffuse in location and extended in time also affect task execution and can affect brain mapping. This may in part reflect fluctuating levels of attention, engagement, or motivation during testing.

Genetic Characterization of Streptococcus pyogenes emm89 Strains Isolated in Japan From 2011 to 2019.

Invasive infection caused by Streptococcus pyogenes emm89 strains has been increasing in several countries linked to a recently emergent clade of emm89 strains, designated clade 3. In Japan, the features of emm89 S. pyogenes strains, such as clade classification, remains unknown. In this study, we collected emm89 strains isolated from both streptococcal toxic shock syndrome (STSS) (89 STSS isolates) and noninvasive infections (72 non-STSS isolates) in Japan from 2011 to 2019, and conducted whole-genome sequencing and comparative analysis, which resulted in classification of a large majority into clade 3 regardless of disease severity. In addition, invasive disease-associated factors were found among emm89 strains, including mutations of control of virulence sensor, and absence of the hylP1 gene encoding hyaluronidase. These findings provide new insights into genetic features of emm89 strains.

Bicarbonate and unsaturated fatty acids enhance capsular polysaccharide synthesis gene expression in oral streptococci, Streptococcus anginosus.

We recently reported on the capsular polysaccharide (CP) synthesis (cps) genes of the oral streptococci, Streptococcus anginosus. In this study, we investigate the effects of carbon dioxide (CO2), bicarbonate (HCO3-) and unsaturated fatty acids (UFAs) on CP synthesis of S. anginosus. We found that CP production increased when bacteria were exposed to high concentrations of CO2. This increase was similarly observed in the presence of sodium bicarbonate (NaHCO3) under atmospheric condition. Since ectopic expression of carbonic anhydrase, which converts CO2 to HCO3-, eliminated the requirement for CO2 in CP production and growth of S. anginosus lacking this enzyme, it seemed that HCO3- is an essential factor for CP production. Furthermore, UFAs also stimulated the CP production. Promoter-reporter assay and quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis confirmed that stimulation of CP production occurs at the transcription level. The results of the promoter assays suggest that the expression and stimulation of cps genes by HCO3- or UFAs require the cpsA gene, which is located in the first position of the cps operon. With respect to the relationship between HCO3-and UFAs, HCO3- may stimulate UFA synthesis pathway at transcription level. Therefore, it is possible that UFAs are definitive signals for the CP production in S. anginosus.

Background Factors Determining the Introduction and Dosage of Insulin in Women With Gestational Diabetes Mellitus.

BACKGROUND: Gestational diabetes mellitus (GDM) is a risk for perinatal complication, and appropriate diagnosis of and intervention in this condition are important. This study aimed to identify patient factors associated with introduction and dosage of insulin, which is the main drug for treatment of GDM. METHODS: In total, 114 patients who had been diagnosed with GDM at our hospital were included in this study. We retrospectively collected clinical parameters of GDM patients, including how many times positive glucose tolerance test results were obtained, whether insulin was introduced, dosage of insulin, body weight, and infant weight. Background factors differing between the insulin introduction and non-introduction groups of GDM patients and parameters associated with the insulin dosage were analyzed. RESULTS: Insulin was introduced in 51 GDM patients (45%). In the insulin introduction group, the six-divided diet was less common and the 75-g glucose tolerance test result was positive a significantly greater number of times compared with the non-introduction group. The factor associated with the insulin introduction status was the number of positive 75-g glucose tolerance test results (odds ratio (OR) 2.04, 95% confidence interval (CI): 1.09 - 3.81, P value = 0.025). In addition, the insulin dosage was found to positively correlate with body weight in the non-pregnant state (P value = 0.005). CONCLUSIONS: The six-divided diet was effective for blood glucose control in GDM women. A positive correlation found between the insulin dosage and body weight in the non-pregnant state suggests the importance of proper pre-pregnancy body weight control.

Risk Factors of Hypoglycemic Encephalopathy and Prolonged Hypoglycemia in Patients With Severe Hypoglycemia.

BACKGROUND: The aim of the study was to investigate risk factors of hypoglycemic encephalopathy (HE) in patients with severe hypoglycemia. METHODS: We retrospectively enrolled patients with severe hypoglycemia who were transported to the emergency department in an ambulance. We defined severe hypoglycemia as plasma glucose level < 60 mg/dL (or capillary levels < 50 mg/dL). HE was defined as severe hypoglycemia with altered level of consciousness (Glasgow coma scale < 12) and prolonged HE as coma or stupor lasting > 24 h after glucose administration. We compared several parameters between patients with and without HE and between prolonged and recovered patients. RESULTS: Included were 173 patients with severe hypoglycemia; of them, 94 were diagnosed with HE, with 12 of them prolonged HE. Glucose level in HE patients was lower than that in those without HE (P < 0.001). Moreover, we noted a significant difference in glucose levels between the prolonged and recovered groups. Furthermore, body temperature was higher in prolonged versus recovered patients (P = 0.0017). CONCLUSION: Blood glucose level may be correlated with severity of altered level of consciousness. In addition, body temperature may be related to coma or prolonged stupor.

Learning process for identifying different types of communication via repetitive stimulation: feasibility study in a cultured neuronal network.

It is well known that various types of information can be learned and memorized via repetitive training. In brain information science, it is very important to determine how neuronal networks comprising neurons with fluctuating characteristics reliably learn and memorize information. The aim of this study is to investigate the learning process in cultured neuronal networks and to address the question described above. Previously, we reported that the spikes resulting from stimulation at a specific neuron propagate as a cluster of excitation waves called spike wave propagation in cultured neuronal networks. We also reported that these waves have an individual spatiotemporal pattern that varies according to the type of neuron that is stimulated. Therefore, different spike wave propagations can be identified via pattern analysis of spike trains at particular neurons. Here, we assessed repetitive stimulation using intervals of 0.5 and 1.5 ms. Subsequently, we analyzed the relationship between the repetition of the stimulation and the identification of the different spike wave propagations. We showed that the various spike wave propagations were identified more precisely after stimulation was repeated several times using an interval of 1.5 ms. These results suggest the existence of a learning process in neuronal networks that occurs via repetitive training using a suitable interval.