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Public acceptance of genetically modified crops engineered with Bacillus thuringiensis (Bt) insecticidal protein genes (BT-GMCs), which confer resistance to various lepidopteran insect pests, is generally lacking. As a major concern over BT-GMCs is the allergenicity of insecticidal proteins, alleviating safety concerns should help increase public acceptance. In this study, three lepidopteran-specific Bt toxins, Cry1Aa, Cy1Ab, and Cry1Ac, were treated with simulated digestive fluids under various conditions. Western blotting using antiserum raised against individual segments (α-helices of domain I and β-sheets of domains II and III) of Cry1Aa showed that digestion produces a variety of polypeptides. In particular, the transmembrane α4–α5 of domain I, which may retain the ability to form pores, was the most resistant to digestion. Intact Cry1A toxins and these digests were then applied to RBL-2H3 cultured rat mast cells to determine whether the toxins directly induce histamine release. However, fluorescence microscopy revealed no specific binding of Cry1A toxins to RBL-2H3 cultured rat mast cells. In addition, neither the OPA method nor HPLC analysis detected significant histamine release from mast cells treated with Cry1A toxins and these digests. Our results provide important data supporting the safety of Cry1A toxins and potentially BT-GMCs.

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Wildfires pose a major health risk for humans, wildlife, and domestic animals. We previously discovered pathophysiologic parallels between domestic cats with naturally occurring smoke inhalation and thermal burn injuries and human beings with similar injuries; these were characterized by transient myocardial thickening, cardiac troponin I elevation and formation of intracardiac thrombosis. While the underlying mechanisms remain unclear, results from murine models suggest that platelet priming and activation may contribute to a global hypercoagulable state and thrombosis. Herein, we evaluated and compared the degree of platelet activation, platelet response to physiologic agonists and levels of platelet-derived microvesicles (PDMV) in 29 cats with naturally occurring wildfire thermal injuries (WF), 21 clinically healthy cats with subclinical hypertrophic cardiomyopathy (HCM) and 11 healthy cats without HCM (CC). We also quantified and compared circulating PDMVs in WF cats to CC cats. In addition, we examined the association between thrombotic events, severity of burn injuries, myocardial changes, and the degree of platelet activation in cats exposed to wildfires. Flow cytometric detection of platelet surface P-selectin expression showed that WF cats had increased platelet response to adenosine diphosphate (ADP) and thrombin compared to the two control groups indicating the presence of primed platelets in circulation. In addition, cats in the WF group had increased circulating levels of PDMV, characterized by increased phosphatidylserine on the external leaflet. Cats in the WF group with documented intracardiac thrombosis had elevated platelet activation and platelet priming in the presence of ADP. While high dose arachidonic acid (AA) mostly resulted in platelet inhibition, persistent response to AA was noted among cats in the WF group with intracardiac thrombosis. Univariate and multiple logistic regression analyses demonstrated that increased platelet response to AA was independently associated with thrombotic events. This is the first study reporting the significant association between platelet priming and intracardiac thrombosis in domestic cats with naturally occurring wildfire-related injuries and smoke inhalation. Further studies are required to delineate additional mechanisms between inflammation and thrombosis, especially regarding platelet primers and the cyclooxygenase pathway.One sentence summaryPlatelet activation and shedding of platelet-derived microvesicles due to platelet priming is present following naturally occurring wildfire smoke exposure and thermal burn injuries in a population of domestic cats.

Background and Objectives: Bedridden patients are at a high risk of venous thromboembolism (VTE). Passive devices such as elastic compression stockings and intermittent pneumatic compression are common. Leg exercise apparatus (LEX) is an active device designed to prevent VTE by effectively contracting the soleus muscle and is therefore expected to be effective in preventing disuse of the lower limbs. However, few studies have been conducted on the kinematic properties of LEX. Therefore, this study aimed to compare the exercise characteristics of LEX with those of an ergometer, which is commonly used as a lower-limb exercise device, and examine its effect on the two domains of muscle activity and circulatory dynamics. Materials and Methods: This study used a crossover design in which each participant performed both exercises to evaluate the exercise characteristics of each device. Fifteen healthy adults performed exercises with LEX and an ergometer (Terasu Erugo, SDG Co., Ltd., Tokyo, Japan) for 5 min each and rested for 10 min after each exercise. Muscle activity was measured using surface electromyography (Clinical DTS, Noraxon, Scottsdale, AZ, USA), and circulatory dynamics were recorded using a non-invasive impedance cardiac output meter (Physioflow Enduro, Manatec Biomedical, Paris, France). The primary outcome was the mean percentage of maximum voluntary contraction (%MVC) of the soleus muscle during exercise. Results: The mean %MVC of the soleus muscle was significantly higher in the LEX group, whereas no significant differences were observed across the periods and sequences. Heart rate, stroke volume, and cardiac output increased during exercise and decreased thereafter; however, the differences between the devices were not significant. Conclusions: LEX may not only have a higher thromboprophylaxis effect, but also a higher effect on preventing muscle atrophy as a lower-extremity exercise device. Additionally, LEX could potentially be used safely in patients who need to be monitored for changes in circulatory dynamics.

Background/Objectives: Preoperative muscle atrophy leads to persistent gait abnormalities in patients undergoing total hip arthroplasty (THA). Efficient motor learning of the gluteus medius is crucial for their recovery. In this study, a single-joint hybrid assistive limb (HAL) was developed to assist hip abduction. We aimed to evaluate the muscle activity and safety of this device during hip abduction in healthy adults. Methods: Ten healthy adults (five males and five females; mean age, 40.7 years) with no hip disorders performed one set of 30 repetitions of side-lying hip abduction under three conditions: without HAL (pre-HAL), with HAL, and without HAL (post-HAL). Muscle activities of the gluteus medius, gluteus maximus, tensor fasciae latae, rectus femoris, and biceps femoris (expressed as percentage of maximum voluntary contraction [%MVC]); vital signs; hip visual analog scale (VAS); and hip abduction and flexion angles were assessed. The mean values were compared among the conditions. Results: The %MVC of the gluteus medius significantly increased from 52% (pre-HAL) to 75.4% (HAL) and then decreased slightly to 61.6% (post-HAL). No other muscle groups showed significant changes. Vital signs and hip VAS scores showed no significant variation. Although no significant differences were found in the hip abduction and flexion angles, a reduction in the hip flexion angle was observed in the HAL and post-HAL conditions. Conclusions: The hip abduction HAL effectively and safely enhanced gluteus medius activity. Reduction in the hip flexion angle during HAL and post-HAL suggests the possibility of appropriate abduction movements and motor learning effects. [ABSTRACT FROM AUTHOR]

Simple Summary: The soil bacterium Bacillus thuringiensis (Bt) produces protein crystals during sporulation. Cry toxins, a major component of these protein crystals, are insecticidal proteins with pore-forming activity. Cry toxins are environmentally friendly biopesticides used to create Bt-incorporating genetically modified crops (BT-GMCs) that confer resistance to insect pests. However, public acceptance of BT-GMCs is lacking. A major concern over BT-GMCs is the allergenicity of Cry toxins. Specifically, the pore-forming activity of Cry toxins may directly induce intestinal mast cells to release histamine. In this study, three lepidopteran-specific Cry toxins, Cry1Aa, Cy1Ab, and Cry1Ac, were treated with simulated digestive fluids under different conditions. Cry1A toxins exhibited differing sensitivity to simulated digestive fluids, and digests of Cry1A toxins contained the α4–α5 helical hairpin of domain I, which is thought to be the transmembrane domain. To determine whether Cry1A toxins directly induce histamine release, intact and digested Cry1A toxins were applied to RBL-2H3 cultured rat mast cells. However, no histamine secretion from these mast cells was detected, even when samples were analyzed using HPLC. The results of our study provide important data supporting the safety of Cry1A toxins and potentially BT-GMCs, and reducing anxiety over their safety should help increase public acceptance. Public acceptance of genetically modified crops engineered with Bacillus thuringiensis (Bt) insecticidal protein genes (BT-GMCs), which confer resistance to various lepidopteran insect pests, is generally lacking. As a major concern over BT-GMCs is the allergenicity of insecticidal proteins, alleviating safety concerns should help increase public acceptance. In this study, three lepidopteran-specific Bt toxins, Cry1Aa, Cy1Ab, and Cry1Ac, were treated with simulated digestive fluids under various conditions. Western blotting using antiserum raised against individual segments (α-helices of domain I and β-sheets of domains II and III) of Cry1Aa showed that digestion produces a variety of polypeptides. In particular, the transmembrane α4–α5 of domain I, which may retain the ability to form pores, was the most resistant to digestion. Intact Cry1A toxins and these digests were then applied to RBL-2H3 cultured rat mast cells to determine whether the toxins directly induce histamine release. However, fluorescence microscopy revealed no specific binding of Cry1A toxins to RBL-2H3 cultured rat mast cells. In addition, neither the OPA method nor HPLC analysis detected significant histamine release from mast cells treated with Cry1A toxins and these digests. Our results provide important data supporting the safety of Cry1A toxins and potentially BT-GMCs. [ABSTRACT FROM AUTHOR]

We developed a serious game with avatars driven by empirically-based algorithms of suspect behavior to train participants in using the Evidence Framing Matrix (EFM), a component of the SUE technique. Ninety-six participants were allocated into four groups (i.e. Control, Training, Feedback, and Training & Feedback) for two interviews. EFM-trained participants used the EFM more than untrained participants in the first interview. Receiving feedback improved EFM use in the second interview. Theoretical training plus feedback made participants use within-statement (in)consistencies more to evaluate avatar statements. The results suggested the potential for using algorithm-driven suspect avatars in investigative interview training. [ABSTRACT FROM AUTHOR]

Abstract CUX2 gene encodes a transcription factor that controls neuronal proliferation, dendrite branching and synapse formation, locating at the epilepsy-associated chromosomal region 12q24 that we previously identified by a genome-wide association study (GWAS) in Japanese population. A CUX2 recurrent de novo variant p.E590K has been described in patients with rare epileptic encephalopathies and the gene is a candidate for the locus, however the mutation may not be enough to generate the genome-wide significance in the GWAS and whether CUX2 variants appear in other types of epilepsies and physiopathological mechanisms are remained to be investigated. Here in this study, we conducted targeted sequencings of CUX2, a paralog CUX1 and its short isoform CASP harboring a unique C-terminus on 271 Japanese patients with a variety of epilepsies, and found that multiple CUX2 missense variants, other than the p.E590K, and some CASP variants including a deletion, predominantly appeared in patients with temporal lobe epilepsy (TLE). The CUX2 variants showed abnormal localization in human cell culture analysis. While wild-type CUX2 enhances dendritic arborization in fly neurons, the effect was compromised by some of the variants. Cux2- and Casp-specific knockout mice both showed high susceptibility to kainate, increased excitatory cell number in the entorhinal cortex, and significant enhancement in glutamatergic synaptic transmission to the hippocampus. CASP and CUX2 proteins physiologically bound to each other and co-expressed in excitatory neurons in brain regions including the entorhinal cortex. These results suggest that CUX2 and CASP variants contribute to the TLE pathology through a facilitation of excitatory synaptic transmission from entorhinal cortex to hippocampus.

IntroductionIn forensic settings interviewers are advised to ask as many open-ended questions as possible. However, even experts may have difficulty following this advice potentially negatively impacting an investigation. Here, we sought to investigate how emotions and psychophysiological parameters are associated with question formulation in real time in an ongoing (simulated) child sexual abuse (CSA) interview.MethodIn a experimental study, psychology students (N = 60, Mage = 22.75) conducted two interviews with child avatars, while their emotions (anger, sadness, disgust, surprise and relief), GSR and heart rate (HR) were registered.ResultsFirst, we found that general emotionality related to CSA and perceived realness of the avatars was associated with stronger overall emotional reactions. Second, we found that closed (vs. open) questions were preceded by more facially observable anger, but not disgust, sadness, surprise or relief. Third, closed (vs. open) questions were preceded by higher GSR resistance and lower heart rate.DiscussionResults suggest for the first time that emotions and psychophysiological states can drive confirmation bias in question formulation in real time in CSA.

Previous research has shown that simulated child sexual abuse (CSA) interview training using avatars paired with feedback and modeling improves interview quality. However, to make this approach scalable, the classification of interviewer questions needs to be automated. We tested an automated question classification system for these avatar interviews while also providing automated interventions (feedback and modeling) to improve interview quality. Forty-two professionals conducted two simulated CSA interviews online and were randomly provided with no intervention, feedback, or modeling after the first interview. Feedback consisted of the outcome of the alleged case and comments on the quality of the interviewer’s questions. Modeling consisted of learning points and videos illustrating good and bad questioning methods. The total percentage of agreement in question coding between human operators and the automated classification was 72% for the main categories (recommended vs. not recommended) and 52% when 11 subcategories were considered. The intervention groups improved from first to second interview while this was not the case in the no intervention group (intervention x time: p = 0.007, ηp2 = 0.28). Automated question classification worked well for classifying the interviewers’ questions allowing interventions to improve interview quality.

Previous research has shown that simulation training using avatars with repeated feedback improves child sexual abuse interview quality. The present study added a hypothesis-testing intervention and examined if the combination of two interventions, feedback and hypothesis-testing, would improve interview quality compared to no intervention and to either intervention alone. Eighty-one Chinese university students were randomly assigned to a control, feedback, hypothesis-testing, or the combination of feedback and hypothesis-testing group and conducted five simulated child sexual abuse interviews online. Depending on the assigned group, feedback on the outcome of the cases and question types used in the interview were provided after each interview, and/or the participants built hypotheses based on preliminary case information before each interview. The combined interventions group and feedback group showed a higher proportion of recommended questions and correct details from the 3rd interview onward compared to the hypothesis-building and control groups. The difference between the number of correct conclusions was not significant. hypothesis-testing alone exacerbated the use of non-recommended questions over time. The results show that hypothesis-testing may impact question types used negatively but not when combined with feedback. The potential reasons for hypothesis-testing alone not being effective and the differences between the present and previous studies were discussed.

Wildfires pose a major health risk for humans, wildlife, and domestic animals. We previously discovered pathophysiologic parallels between domestic cats with naturally occurring smoke inhalation and thermal burn injuries and human beings with similar injuries; these were characterized by transient myocardial thickening, cardiac troponin I elevation and formation of intracardiac thrombosis. While the underlying mechanisms remain unclear, results from murine models suggest that platelet priming and activation may contribute to a global hypercoagulable state and thrombosis. Herein, we evaluated and compared the degree of platelet activation, platelet response to physiologic agonists and levels of platelet-derived microvesicles (PDMV) in 29 cats with naturally occurring wildfire thermal injuries (WF), 21 clinically healthy cats with subclinical hypertrophic cardiomyopathy (HCM) and 11 healthy cats without HCM (CC). We also quantified and compared circulating PDMVs in WF cats to CC cats. In addition, we examined the association between thrombotic events, severity of burn injuries, myocardial changes, and the degree of platelet activation in cats exposed to wildfires. Flow cytometric detection of platelet surface P-selectin expression showed that WF cats had increased platelet response to adenosine diphosphate (ADP) and thrombin compared to the two control groups indicating the presence of primed platelets in circulation. In addition, cats in the WF group had increased circulating levels of PDMV, characterized by increased phosphatidylserine on the external leaflet. Cats in the WF group with documented intracardiac thrombosis had elevated platelet activation and platelet priming in the presence of ADP. While high dose arachidonic acid (AA) mostly resulted in platelet inhibition, persistent response to AA was noted among cats in the WF group with intracardiac thrombosis. Univariate and multiple logistic regression analyses demonstrated that increased platelet response to AA was independently associated with thrombotic events. This is the first study reporting the significant association between platelet priming and intracardiac thrombosis in domestic cats with naturally occurring wildfire-related injuries and smoke inhalation. Further studies are required to delineate additional mechanisms between inflammation and thrombosis, especially regarding platelet primers and the cyclooxygenase pathway.One Sentence SummaryPlatelet activation and shedding of platelet-derived microvesicles due to platelet priming is present following naturally occurring wildfire smoke exposure and thermal burn injuries in a population of domestic cats.

Background and Objectives: Bedridden patients are at a high risk of venous thromboembolism (VTE). Passive devices such as elastic compression stockings and intermittent pneumatic compression are common. Leg exercise apparatus (LEX) is an active device designed to prevent VTE by effectively contracting the soleus muscle and is therefore expected to be effective in preventing disuse of the lower limbs. However, few studies have been conducted on the kinematic properties of LEX. Therefore, this study aimed to compare the exercise characteristics of LEX with those of an ergometer, which is commonly used as a lower-limb exercise device, and examine its effect on the two domains of muscle activity and circulatory dynamics. Materials and Methods: This study used a crossover design in which each participant performed both exercises to evaluate the exercise characteristics of each device. Fifteen healthy adults performed exercises with LEX and an ergometer (Terasu Erugo, SDG Co., Ltd., Tokyo, Japan) for 5 min each and rested for 10 min after each exercise. Muscle activity was measured using surface electromyography (Clinical DTS, Noraxon, Scottsdale, AZ, USA), and circulatory dynamics were recorded using a non-invasive impedance cardiac output meter (Physioflow Enduro, Manatec Biomedical, Paris, France). The primary outcome was the mean percentage of maximum voluntary contraction (%MVC) of the soleus muscle during exercise. Results: The mean %MVC of the soleus muscle was significantly higher in the LEX group, whereas no significant differences were observed across the periods and sequences. Heart rate, stroke volume, and cardiac output increased during exercise and decreased thereafter; however, the differences between the devices were not significant. Conclusions: LEX may not only have a higher thromboprophylaxis effect, but also a higher effect on preventing muscle atrophy as a lower-extremity exercise device. Additionally, LEX could potentially be used safely in patients who need to be monitored for changes in circulatory dynamics. [ABSTRACT FROM AUTHOR]

Many causative genes are known for epileptic or developmental and epileptic encephalopathies (EE/DEE) yet a genetic diagnosis cannot be made for many patients. Here, the authors analyse whole exome sequencing data from a Japanese case−control cohort to identify common, rare and ultra-rare coding variants associated with EE/DEE.

Although previous research has confirmed the effectiveness of simulated child sexual abuse interviews with feedback, its validation is limited to Western contexts and face-to-face settings. The present study aims to extend this research to non-Western and online/remote training conditions. Thirty-two Japanese undergraduate students were randomly assigned to a control or feedback group. The feedback group conducted a set of six online simulated child sexual abuse interviews while receiving feedback after each interview in an attempt to improve the quality of their questioning style. The feedback consisted of the outcome of the alleged cases and the quality of the questions asked in the interviews. The control group conducted the interviews without feedback. The feedback (vs. control) increased the proportion of recommended questions (first interview: 45%; last interview: 65% vs. first: 43%; last: 42%, respectively) by using fewer not-recommended questions and eliciting fewer incorrect details. Furthermore, only participants in the feedback group (7 out of 17) demonstrated a reliable change in the proportion of recommended questions. The present study explores the efficacy of simulated interview training with avatars in a different cultural setting and in the context of remote administration. The differences between the present study and previous research are discussed in light of cultural and logistical aspects.

Selective dorsal rhizotomy (SDR) has been used to treat children with spastic cerebral palsy (CP), and its beneficial effect on quality of life and ambulation has been confirmed in long-term follow-up studies. However, the role of SDR in the treatment of spasticity in patients with hereditary spastic paraplegia (HSP) and related disorders is not well-established. Here, we report the first patient with the ZC4H2 variant who underwent SDR to treat spastic paraplegia. Abnormal gait was discovered during a regular checkup at the age of 3 years and 9 months, and she was diagnosed with spastic paraplegia. She was heterozygous for the ZC4H2 variant and underwent SDR at the age of 5 years and 11 months, which alleviated the spasticity. The patient underwent inpatient postoperative rehabilitation for 4 months and continued outpatient physiotherapy after discharge. The Gross Motor Function Measure-88 score and maximum walking speed decreased transiently 1 month postoperatively, but gradually recovered, and continuously improved 6 months postoperatively. SDR and postoperative intensive rehabilitation were effective in improving motor and walking functions up to 6 months after surgery, although long-term follow-up is needed to draw conclusions. [ABSTRACT FROM AUTHOR]

Simulated avatar interview training has been proven to be effective in improving child sexual abuse interview quality. However, the topic of perceived realism of the avatars and whether they cause emotional reactions has not been previously investigated. Such reactions could affect both learning from the interview simulations as well as how actual interviews are conducted. We wanted to understand whether participants perceive allegedly sexually abused child avatars as realistic and how they emotionally respond to avatars revealing they were actually abused vs. not-abused. Psychology students and recent graduates (N = 30, Mage = 27.9 years) watched eight avatars (four boys, four girls, four with a CSA and four with a no-CSA scenario) providing a series of details about what had happened. Before and after observing each avatar, the participants' emotional reactions and perceived realness of the avatars were measured. Also, during each observation, the participant's facial expressions were recorded. The participants self-reported more negative (anger, sadness, disgust) and more positive (relief) emotions to confirmed CSA and disconfirmed CSA scenarios, respectively, while results for facially expressed emotions were less clear. Higher general emotionality related to CSA and higher perceived realness of the avatars made the differences generally stronger. [ABSTRACT FROM AUTHOR]

Introduction Lipids have various roles in cells, including that of energy source, membrane structural component, signaling molecule, and platform for the recruitment and activation of enzymes (1). Their backbone (glycerophospholipids, [...], ATP11A translocates phosphatidylserine (PtdSer), but not phosphatidylcholine (PtdCho), from the outer to the inner leaflet of plasma membranes, thereby maintaining the asymmetric distribution of PtdSer. Here, we detected a de novo heterozygous point mutation of ATP11A in a patient with developmental delays and neurological deterioration. Mice carrying the corresponding mutation died perinatally of neurological disorders. This mutation caused an amino acid substitution (Q84E) in the first transmembrane segment of ATP11A, and mutant ATP11A flipped PtdCho. Molecular dynamics simulations revealed that the mutation allowed PtdCho binding at the substrate entry site. Aberrant PtdCho flipping markedly decreased the concentration of PtdCho in the outer leaflet of plasma membranes, whereas sphingomyelin (SM) concentrations in the outer leaflet increased. This change in the distribution of phospholipids altered cell characteristics, including cell growth, cholesterol homeostasis, and sensitivity to sphingomyelinase. Matrix-assisted laser desorption ionization-imaging mass spectrometry (MALDI-IMS) showed a marked increase of SM levels in the brains of Q84E-knockin mouse embryos. These results provide insights into the physiological importance of the substrate specificity of plasma membrane flippases for the proper distribution of PtdCho and SM.