A type II toxin-antitoxin system, ECs3274-ECs3275, in enterohemorrhagic Escherichia coli O157.

The toxin-antitoxin (TA) genetic module controls various bacterial events. Novel toxins with different functions are still being discovered. This study aimed to determine whether the ECs3274-ECs3275 gene pair encoded by enterohemorrhagic Escherichia coli O157 functions as a TA system. To characterize this putative TA system, we analyzed the growth of E. coli expressing ECs3274, ECs3275, or both; the interaction between ECs3274 and ECs3275 using bacterial adenylate cyclase two-hybrid assays; and the DNA-binding ability of ECs3274 using gel-mobility shift assays. We observed that the ECs3274 antitoxin interacted with the ECs3275 toxin, was destabilized by Lon protease, and repressed its promoter activity via its helix-turn-helix (HTH) motif. These properties are consistent with those of typical type II TA antitoxins. Interestingly, ECs3275 has an HTH motif not observed in other TA toxins and is necessary for ECs3275 toxicity, suggesting that ECs3275 may exert its toxicity by regulating the expression of specific genes.

The use of concomitant medications on nephrotoxicity associated with teicoplanin: A retrospective observational study.

BACKGROUND: Teicoplanin (TEIC) is a nephrotoxic agent. However, little is known about the effects of concomitant medications on nephrotoxicity. In this study, we investigated the effects of concomitant drugs on nephrotoxicity. METHODS: A retrospective observational case-control study was conducted on patients (≥18 years) who started TEIC at the Tokyo Dental College, Ichikawa General Hospital, between January 2013 and April 2023. The primary outcome was nephrotoxicity, defined as an increase in serum creatinine levels of ≥50 % or ≥0.5 mg/dL from baseline. Logistic regression analysis was used to determine the risk factors for nephrotoxicity associated with TEIC. In addition, we investigated the relationship between nephrotoxicity and predicted free TEIC concentrations. RESULTS: Of 305 patients, 43 (14.1 %) developed nephrotoxicity. The multivariate logistic regression analysis identified that serum albumin (odds ratio [OR] = 0.50, 95 % confidence interval [CI] 0.27-0.89, p = 0.02), concomitant use of loop diuretics (OR = 2.22, 95 % CI 1.10-4.59, p = 0.03), antivirals (OR = 3.24, 95 % CI 1.32-7.62, p < 0.01), and vasopressors (OR = 2.57, 95 % CI 1.10-5.78, p = 0.03) were the associated risk factors for nephrotoxicity in patients administered with TEIC. In 216 patients, predicted TEIC concentrations were 3.6 [interquartile range (IQR), 2.6-4.9] µg/mL in the nephrotoxicity group versus 3.6 [IQR, 2.5-4.7] µg/mL in the non-nephrotoxicity group, with no significant difference (p = 0.69). CONCLUSION: Our results indicate the importance of modifying the concomitant use of loop diuretics, antivirals, and vasopressors.

Customized Visual Discrimination Digital Therapy According to Visual Field Defects in Chronic Stroke Patients.

BACKGROUND AND PURPOSE: Visual perceptual learning (VPL) may improve visual field defects (VFDs) after chronic stroke, but the optimal training duration and location remain unknown. This prospective study aimed to determine the efficacy of 8 weeks of VFD-customized visual discrimination training in improving poststroke VFDs. METHODS: Prospectively enrolled patients with poststroke VFDs initially received no training for 8 weeks (no-training phase). They subsequently underwent our customized VPL program that included orientation-discrimination tasks in individualized blind fields and central letter-discrimination tasks three times per week for 8 weeks (training phase). We analyzed the luminance detection sensitivity and deviation as measured using Humphrey visual field tests before and after the no-training and training phases. The vision-related quality of life was assessed at baseline and at a 16-week follow-up using the National Eye Institute Visual Function Questionnaire-25 (NEI-VFQ-25). RESULTS: Changes in mean total deviation (MTD) scores were greater during the training phase than during the no-training phase (defective hemifield, p=0.002; whole field, p=0.004). The MTD scores improved during the training phase (defective hemifield, p=0.004; whole field, p=0.016), but not during the no-training phase (defective hemifield, p=0.178; whole field, p=0.178). The difference between the improved and worsened areas (≥6 dB changes in luminance detection sensitivity) was greater during the training phase than during the no-training phase (p=0.009). The vision-specific social functioning subscore of the NEI-VFQ-25 improved after the 16-week study period (p=0.040). CONCLUSIONS: Our 8-week VFD-customized visual discrimination training protocol may effectively improve VFDs and vision-specific social functioning in chronic stroke patients.

Effect of preoperative physical activity on postoperative outcomes in esophageal cancer.

[Purpose] This study aimed to examine the characteristics of preoperative physical activity and its impact on the postoperative period in patients who underwent surgery for esophageal cancer. [Participants and Methods] The participants were 30 patients who were diagnosed with esophageal cancer, underwent surgery, and fulfilled their conditions. Preoperative physical activity was measured using the step count, and metabolic equivalents as the amount of physical activity. We examined the relationships between preoperative step count and METs, patient demographics, treatment-related factors, preoperative physical function, and activities of daily living. Moreover, we examined the relationships of preoperative step count and METs with postoperative mobilization, physical activity, physical function, and activities of daily living. [Results] Preoperative step count was related to age, Glasgow prognostic score, and preoperative functional independence and associated with step count on postoperative days 7-13, METs on postoperative days 7-9, 6-min walking distance, and functional independence measures at discharge. [Conclusion] Improving the nutritional status and increasing preoperative physical activity by walking for esophageal cancer may help improve physical activity after postoperative day 7, exercise tolerance, and activities of daily after discharge.

Unsupervised learning as a computational principle works in visual learning of natural scenes, but not of artificial stimuli.

The question of whether we learn exposed visual features remains a subject of controversy. A prevalent computational model suggests that visual features frequently exposed to observers in natural environments are likely to be learned. However, this unsupervised learning model appears to be contradicted by the significant body of experimental results with human participants that indicates visual perceptual learning (VPL) of visible task-irrelevant features does not occur with frequent exposure. Here, we demonstrate a resolution to this controversy with a new finding: Exposure to a dominant global orientation as task-irrelevant leads to VPL of the orientation, particularly when the orientation is derived from natural scene images, whereas VPL did not occur with artificial images even with matched distributions of local orientations and spatial frequencies to natural scene images. Further investigation revealed that this disparity arises from the presence of higher-order statistics derived from natural scene images-global structures such as correlations between different local orientation and spatial frequency channels. Moreover, behavioral and neuroimaging results indicate that the dominant orientation from these higher-order statistics undergoes less attentional suppression than that from artificial images, which may facilitate VPL. Our results contribute to resolving the controversy by affirming the validity of unsupervised learning models for natural scenes but not for artificial stimuli. They challenge the assumption that VPL occurring in everyday life can be predicted by laws governing VPL for conventionally used artificial stimuli.

Prostacyclin synthase deficiency exacerbates systemic inflammatory responses in lipopolysaccharide-induced septic shock in mice.

OBJECTIVES: Sepsis is a systemic inflammatory disorder characterized by life-threateningorgan dysfunction resulting from a dysregulated host response to infection. Prostacyclin (PGI2) is a bioactive lipid produced by PGI synthase (PGIS) and is known to play important roles in inflammatory reactions as well as cardiovascular regulation. However, little is known about the roles of PGIS and PGI2 in systemic inflammatory responses such as septic shock. METHODOLOGY: Systemic inflammation was induced by intraperitoneal injection of 5 mg/kg lipopolysaccharide (LPS) in wild type (WT) or PGIS knockout (KO) mice. Selexipag, a selective PGI2 receptor (IP) agonist, was administered 2 h before LPS injection and again given every 12 h for 3 days. RESULTS: Intraperitoneal injection of LPS induced diarrhea, shivering and hypothermia. These symptoms were more severe in PGIS KO mice than in WT micqe. The expression of Tnf and Il6 genes was notably increased in PGIS KO mice. In contrast, over 95% of WT mice survived 72 h after the administration of LPS, whereas all of the PGIS KO mice had succumbed by that time. The mortality rate of LPS-administrated PGIS KO mice was improved by selexipag administration. CONCLUSION: Our study suggests that PGIS-derived PGI2 negatively regulates LPS-induced symptoms via the IP receptor. PGIS-derived PGI2-IP signaling axis may be a new drug target for systemic inflammation in septic shock.

First-night effect reduces the beneficial effects of sleep on visual plasticity and modifies the underlying neurochemical processes.

Individuals experience difficulty falling asleep in a new environment, termed the first night effect (FNE). However, the impact of the FNE on sleep-induced brain plasticity remains unclear. Here, using a within-subject design, we found that the FNE significantly reduces visual plasticity during sleep in young adults. Sleep-onset latency (SOL), an indicator of the FNE, was significantly longer during the first sleep session than the second session, confirming the FNE. We assessed performance gains in visual perceptual learning after sleep and increases in the excitatory-to-inhibitory neurotransmitter (E/I) ratio in early visual areas during sleep using magnetic resonance spectroscopy and polysomnography. These parameters were significantly smaller in sleep with the FNE than in sleep without the FNE; however, these parameters were not correlated with SOL. These results suggest that while the neural mechanisms of the FNE and brain plasticity are independent, sleep disturbances temporarily block the neurochemical process fundamental for brain plasticity.

Learning of the same task subserved by substantially different mechanisms between patients with body dysmorphic disorder and healthy individuals.

It has remained unclear whether individuals with psychiatric disorders involving altered visual processing employ similar neuronal mechanisms during perceptual learning of a visual task. We investigated this question by training patients with body dysmorphic disorder, a psychiatric disorder characterized by distressing or impairing preoccupation with nonexistent or slight defects in one's physical appearance, and healthy controls on a visual detection task for human faces with low spatial frequency components. Brain activation during task performance was measured with functional magnetic resonance imaging before the beginning and after the end of behavioral training. Both groups of participants improved performance on the trained task to a similar extent. However, neuronal changes in the fusiform face area were substantially different between groups such that activation for low spatial frequency faces in the right fusiform face area increased after training in body dysmorphic disorder patients but decreased in controls. Moreover, functional connectivity between left and right fusiform face area decreased after training in patients but increased in controls. Our results indicate that neuronal mechanisms involved in perceptual learning of a face detection task differ fundamentally between body dysmorphic disorder patients and controls. Such different neuronal mechanisms in body dysmorphic disorder patients might reflect the brain's adaptations to altered functions imposed by the psychiatric disorder.

Activated NAD+ biosynthesis pathway induces olaparib resistance in BRCA1 knockout pancreatic cancer cells.

PARP inhibitors have been developed as anti-cancer agents based on synthetic lethality in homologous recombination deficient cancer cells. However, resistance to PARP inhibitors such as olaparib remains a problem in clinical use, and the mechanisms of resistance are not fully understood. To investigate mechanisms of PARP inhibitor resistance, we established a BRCA1 knockout clone derived from the pancreatic cancer MIA PaCa-2 cells, which we termed C1 cells, and subsequently isolated an olaparib-resistant C1/OLA cells. We then performed RNA-sequencing and pathway analysis on olaparib-treated C1 and C1/OLA cells. Our results revealed activation of cell signaling pathway related to NAD+ metabolism in the olaparib-resistant C1/OLA cells, with increased expression of genes encoding the NAD+ biosynthetic enzymes NAMPT and NMNAT2. Moreover, intracellular NAD+ levels were significantly higher in C1/OLA cells than in the non-olaparib-resistant C1 cells. Upregulation of intracellular NAD+ levels by the addition of nicotinamide also induced resistance to olaparib and talazoparib in C1 cells. Taken together, our findings suggest that upregulation of intracellular NAD+ is one of the factors underlying the acquisition of PARP inhibitor resistance.

Cloning and expression profile of the alanine aminotransferase gene from kuruma shrimp Penaeus japonicus exposed to different salinities.

Several crustaceans including shrimps change the amount of specific free amino acids to regulate the osmotic pressure in their bodies. Kuruma shrimp Penaeus japonicus also increases the concentration of alanine (Ala) in the abdominal muscle following the increase of environmental salinity. In the present study, to elucidate the mechanisms of changes in Ala accumulation of kuruma shrimp depending on salinity, we cloned the gene encoding alanine aminotransferase (ALT), an enzyme involved in Ala biosynthesis, and examined its expression profile. It was found that the full-length kuruma shrimp ALT1 cDNA consisted of 3,301 bp, encoding 514 amino acids, and that all amino acid residues important for ALT activity were conserved. Phylogenetic analysis also indicated that the ALT gene cloned in this study was classified as ALT1. Moreover, we examined the expression levels of the ALT1 gene in the abdominal muscle and the hepatopancreas of kuruma shrimp acclimated at 17‰, 34‰, and 40‰ salinities, resulting that the mRNA levels of the ALT1 genes in both tissues of the shrimp acclimated at 40‰ were significantly higher than those at 17‰ for 12 h (p < 0.05). The mRNA levels of the ALT1 gene in the abdominal muscle of the shrimp acclimated for more than 24 h tended to increase following the increase of environmental salinity. These results indicate that ALT1 is responsible for the increase of free Ala concentration in the abdominal muscle of kuruma shrimp to regulate osmotic pressure at high salinity.

3D-printed custom tray for maxillofacial implant assisted partial denture.

This technique presents a new fabrication workflow for a three-dimensional (3D) printed custom tray, which duplicates the morphology of the treatment denture for maxillofacial prostheses using an intraoral scanner, computer-aided design (CAD) software, and a 3D printer. A 70-year-old man underwent reconstruction of segmental mandibulectomy for mandibular osteoblastoma, followed by implant placement and secondary surgery. During the surgical treatment, a treatment denture was fabricated to restore oral function and determine the morphology of the definitive denture. To create the definitive denture with the same morphology as the treatment denture a custom tray was fabricated with the denture morphology after chairside adjustments. The oral cavity was scanned using an intraoral scanner, and the data acquired were imported into general-purpose CAD software, adjusted, and imported into a 3D printer to produce the custom tray. This was fitted into the patient's mouth without any issues, and closed tray impressions were made with impression caps for the locator attachments on the implant body. The morphology of the treatment denture was replicated in the definitive denture by making a silicon impression of the cameo surface at the fabrication of the cast after impression making. In this technique, the morphology of the treatment denture was transferred accurately to the definitive implant partial denture by leveraging existing digital technology. This method represents a practical approach for partial denture fabrication, including maxillofacial defects with complex denture configurations.

First-night effect reduces the beneficial effects of sleep on visual plasticity and modifies the underlying neurochemical processes.

Individuals experience difficulty falling asleep in a new environment, termed the first night effect (FNE). However, the impact of the FNE on sleep-induced brain plasticity remains unclear. Here, using a within-subject design, we found that the FNE significantly reduces visual plasticity during sleep in young adults. Sleep-onset latency (SOL), an indicator of the FNE, was significantly longer during the first sleep session than the second session, confirming the FNE. We assessed performance gains in visual perceptual learning after sleep and increases in the excitatory-to-inhibitory neurotransmitter (E/I) ratio in early visual areas during sleep using magnetic resonance spectroscopy and polysomnography. These parameters were significantly smaller in sleep with the FNE than in sleep without the FNE; however, these parameters were not correlated with SOL. These results suggest that while the neural mechanisms of the FNE and brain plasticity are independent, sleep disturbances temporarily block the neurochemical process fundamental for brain plasticity.

Shift in excitation-inhibition balance underlies perceptual learning of temporal discrimination.

Temporal perceptual learning (TPL) constitutes a unique and profound demonstration of neural plasticity within the brain. Our understanding for the neurometabolic changes associated with TPL on the other hand has been limited in part by the use of traditional fMRI approaches. Since plasticity in the visual cortex has been shown to underlie perceptual learning of visual information, we tested the hypothesis that TPL of an auditory interval involves a similar change in plasticity of the auditory pathway and if so, whether these changes take place in a lower-order sensory-specific brain area such as the primary auditory cortex (A1), or a higher-order modality-independent brain area such as the inferior parietal cortex (IPC). This distinction will inform us of the mechanisms underlying perceptual learning as well as the locus of change as it relates to TPL. In the present study, we took advantage of a new technique: proton magnetic resonance spectroscopy (MRS) in combination with psychophysical measures to provide the first evidence of changes in neurometabolic processing following 5 days of temporal discrimination training. We measured the (E)xcitation-to-(I)nhibition ratio as an index of learning in the right IPC and left A1 while participants learned an auditory two-tone discrimination task. During the first day of training, we found a significant task-related increase in functional E/I ratio within the IPC. While the A1 exhibited the opposite pattern of neurochemical activity, this relationship did not reach statistical significance. After timing performance has reached a plateau, there were no further changes to functional E/I. These findings support the hypothesis that improvements in temporal discrimination relies on neuroplastic changes in the IPC, but it is possible that both areas work synergistically to acquire a temporal interval.

Functional connectivity interacts with visual perceptual learning for visual field recovery in chronic stroke.

A reciprocal relationship between perceptual learning and functional brain changes towards perceptual learning effectiveness has been demonstrated previously; however, the underlying neural correlates remain unclear. Further, visual perceptual learning (VPL) is implicated in visual field defect (VFD) recovery following chronic stroke. We investigated resting-state functional connectivity (RSFC) in the visual cortices associated with mean total deviation (MTD) scores for VPL-induced VFD recovery in chronic stroke. Patients with VFD due to chronic ischemic stroke in the visual cortex received 24 VPL training sessions over 2 months, which is a dual discrimination task of orientation and letters. At baseline and two months later, the RSFC in the ipsilesional, interhemispheric, and contralesional visual cortices and MTD scores in the affected hemi-field were assessed. Interhemispheric visual RSFC at baseline showed the strongest correlation with MTD scores post-2-month VPL training. Notably, only the subgroup with high baseline interhemispheric visual RSFC showed significant VFD improvement following the VPL training. The interactions between the interhemispheric visual RSFC at baseline and VPL led to improvement in MTD scores and largely influenced the degree of VFD recovery. The interhemispheric visual RSFC at baseline could be a promising brain biomarker for the effectiveness of VPL-induced VFD recovery.

Plasticity-stability dynamics during post-training processing of learning.

Learning continues beyond the end of training. Post-training learning is supported by changes in plasticity and stability in the brain during both wakefulness and sleep. However, the lack of a unified measure for assessing plasticity and stability dynamics during training and post-training periods has limited our understanding of how these dynamics shape learning. Focusing primarily on procedural learning, we integrate work using behavioral paradigms and a recently developed measure, the excitatory-to-inhibitory (E/I) ratio, to explore the delicate balance between plasticity and stability and its relationship to post-training learning. This reveals plasticity-stability cycles during both wakefulness and sleep that enhance learning and protect it from new learning during post-training processing.

Dysfunction of poly (ADP-ribose) glycohydrolase suppresses osteoclast differentiation in RANKL-stimulated RAW264 cells.

Poly (ADP-ribose) glycohydrolase (PARG) is an enzyme that mainly degrades poly (ADP-ribose) (PAR) synthesized by poly (ADP-ribose) polymerase (PARP) family proteins. Although PARG is involved in many biological phenomena, including DNA repair, cell differentiation, and cell death, little is known about the relationship between osteoclast differentiation and PARG. It has also not been clarified whether PARG is a valuable target for therapeutic agents in the excessive activity of osteoclast-related bone diseases such as osteoporosis. In the present study, we examined the effects of PARG inhibitor PDD00017273 on osteoclast differentiation in RANKL-induced RAW264 cells. PDD00017273 induced the accumulation of intracellular PAR and suppressed the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells. PDD00017273 also downregulated osteoclast differentiation marker genes such as Trap, cathepsin K (Ctsk), and dendrocyte expressed seven transmembrane protein (Dcstamp) and protein expression of nuclear factor of activated T cells 1 (NFATc1), a master regulator of osteoclast differentiation. Taken together, our findings suggest that dysfunction of PARG suppresses osteoclast differentiation via the PAR accumulation and partial inactivation of the NFATc1.

Activities of Daily Living, Hypoxemia, and Lymphocytes Score for Predicting Mortality Risk in Patients With Pulmonary TB.

BACKGROUND: A clinically applicable mortality risk prediction system for pulmonary TB may improve treatment outcomes, but no easy-to-calculate and accurate score has yet been reported. The aim of this study was to construct a simple and objective disease severity score for patients with pulmonary TB. RESEARCH QUESTION: Does a clinical score consisting of simple objective factors predict the mortality risk of patients with pulmonary TB? STUDY DESIGN AND METHODS: The data set from our previous prospective study that recruited patients newly diagnosed with pulmonary TB was used for the development cohort. Patients for the validation cohort were prospectively recruited between March 2021 and September 2022. The primary end point was all-cause in-hospital mortality. Using Cox proportional hazards regression, a mortality risk prediction model was optimized in the development cohort. The disease severity score was developed by assigning integral points to each variate. RESULTS: The data from 252 patients in the development cohort and 165 patients in the validation cohort were analyzed, of whom 39 (15.5%) and 17 (10.3%), respectively, died in the hospital. The disease severity score (named the AHL score) included three clinical parameters: activities of daily living (semi-dependent, 1 point; totally dependent, 2 points); hypoxemia (1 point), and lymphocytes (< 720/µL, 1 point). This score showed good discrimination with a C statistic of 0.902 in the development cohort and 0.842 in the validation cohort. We stratified the score into three groups (scores of 0, 1-2, and 3-4), which clearly corresponded to low (0% and 1.3%), intermediate (13.5% and 8.9%), and high (55.8% and 39.3%) mortality risk in the development and validation cohorts. INTERPRETATION: The easy-to-calculate AHL disease severity score for patients with pulmonary TB was able to categorize patients into three mortality risk groups with great accuracy. CLINICAL TRIAL REGISTRATION: University Hospital Medical Information Network Center; No. UMIN000012727 and No. UMIN000043849; URL: www.umin.ac.jp.

Recycling potassium from cow manure compost can replace potassium fertilizers in paddy rice production systems.

The prevalence of K deficiency and negative K balance in rice production increases the demand for K fertilizer. However, the primary source of K fertilizer, potash rock, is limited. Recycling K from cow manure compost (CMC) is a sustainable solution. Nevertheless, the effects of substituting K fertilizer with CMC on rice yield, soil K fertility, and partial K balance (PKB) are not well understood. Therefore, a field experiment with four treatments (control - unfertilized, MNP K - CMC plus NPK fertilizer, MNP ½ K - CMC plus NP and 50 % K fertilizer, and MNP - CMC plus NP fertilizer) was conducted from 2020 to 2022 to study the effects of replacing K fertilizer with K from CMC on rice growth, yield, plant K uptake, soil K fertility, and PKB. The results indicated that K input from CMC exceeded the recommended K fertilizer level, sufficient for optimal rice growth and yield over three growing seasons and plant K uptake in the last two seasons. Plant K uptake increased with total K input and reached a plateau when total K input approached the maximum plant K uptake. In the MNP treatment, PKB was negative in the first two seasons but became positive in the last season, owing to the equivalence between K input from CMC and plant K uptake. Key factors influencing PKB in this treatment were K input from CMC and plant K uptake. Increasing the CMC application rate during the first two seasons could lead to a positive PKB. In this treatment, soil exchangeable K changed correspondingly with PKB, decreasing in the first two seasons but increasing in the last season. Overall, determining the appropriate amount of CMC application for a positive PKB is vital for the sustainability of substituting K fertilizer with K from CMC in paddy rice systems.

A case report of forensic personal identification by transposed teeth and dental treatment marks using multidetector row computed tomography.

Using postmortem computed tomography (CT) images, we achieved personal identification of a body using transposed teeth and dental treatment marks. Transposition of teeth is a rare malpositioning anomaly. CT images can clarify the malpositioning of the teeth's roots, which is difficult to discern from gross observation of the dentition. Because dental hygiene has reduced the incidence of caries in recent years, it might be difficult to use treatment marks for personal identification in the future. Transposed teeth, although rare, provide an important clue to personal identification.

Geometric-relationship specific transfer in visual perceptual learning.

Visual perceptual learning (VPL) is defined as long-term improvement on a visual task as a result of visual experience. In many cases, the improvement is highly specific to the location where the target is presented, which refers to location specificity. In the current study, we investigated the effect of a geometrical relationship between the trained location and an untrained location on transfer of VPL. We found that significant transfer occurs either diagonally or along a line passing the fixation point. This indicates that whether location specificity or location transfer occurs at least partially depends on the geometrical relationship between trained location and an untrained location.