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In dairy cows, low fertility caused by summer heat stress continues into the cooler autumn season. This can be caused by impaired oocyte quality in small growing follicles during summer. Here, we subjected oocyte-cumulus-granulosa complexes (OCGCs) derived from early antral follicles (0.5-1 mm) to in vitro growth (IVG) culture under two different temperature settings (the control and heat shock groups), and evaluated effects of heat exposure on growth and developmental competence of oocytes, factors affecting the developmental competence of oocytes (steroidogenesis of granulosa cells, oxidative stress in oocytes, and cell-to-cell communication between oocytes and somatic cells). Oocyte diameters after culture were smaller in the heat shock group. Although nuclear maturation and cleavage rates were similar between the groups, blastocyst rates were lower in the heat shock group (0.0%) than in the control group (27.7%), and reduced glutathione (GSH) levels in oocytes were lower in the heat shock group. Supplementation of cysteine, which stimulates GSH synthesis, increased GSH level and improved blastocyst rate of heat shocked oocytes (27.9%). These results suggest that heat exposure impairs the growth and developmental competence of oocytes in early antral follicles through GSH depletion, which can induce low fertility during summer and the following autumn., (© 2022. The Author(s).)

Understanding the desires and motivations of children with ADHD is important in helping them thrive. Their inner worlds, however, have not been well captured. The Three Wishes task provides minimal cues and structure to elicit their desires and hopes in an unbiased manner. The wishes of 299 school-aged children with ADHD (193 boys, aged 6-12) were elicited during a research diagnostic assessment. We developed a coding scheme to characterize different aspects of their wishes, including beneficiary, valence, and immediacy. Maslow's hierarchy of needs, adapted to take account of the participants' ages, was used to identify the motivations underlying the children's wishes. As expected, many of the wishes reported were for immediate fulfillment, with many reflecting material desires. Affiliative wishes, highlighting the children's desire for positive interpersonal relationships, were also common. There was some evidence for self-actualization/self-betterment goals and a small number of altruistic wishes. A word cloud presents the content of the children's wishes grouped according to this hierarchy. This study highlights the diversity and typicality of the self-reported needs, desires and hopes of children with ADHD. It also serves as a timely reminder of the value of seeking such information directly from children themselves., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Furukawa, Uchida, Tatsuki, Fitts and Tripp.)

The stigma of infertility negatively affects health, resulting in mental distress and poor quality of life. An appropriate scale is essential to examine the stigma experienced by infertile women and provide adequate interventions. Therefore, we developed a Japanese version of the Infertility Stigma Scale (ISS). After examining the content validity of this scale, we conducted an online survey of women undergoing fertility treatment to test the scale’s structural validity, internal consistency, intra-rater reliability, known-groups validity, convergent validity, and discriminant validity. A total of 254 participants were included in the analysis. The results of confirmatory factor analysis of four factors based on the original scale revealed the optimal fit. Cronbach’s alpha was 0.95 for the total score. Concerning test−retest analysis, the total score of the ISS and subscale had a high Spearman correlation coefficient (ρ = 0.61−0.88, p < 0.001). For convergent validity, the association between the ISS and social support, self-esteem, and family functioning was significantly negatively correlated. The results of the multitrait scaling analysis scale showed that correlations of all items exceeded 0.40, and scaling errors (7/81, 8.6%) were few. The Japanese version of the ISS was confirmed to have acceptable reliability and validity.

The neuroendocrine carcinoma of the gastrointestinal system (GIS-NEC) is a rare but highly malignant neoplasm. We analyzed 115 cases using whole-genome/exome sequencing, transcriptome sequencing, DNA methylation assays, and/or ATAC-seq and found GIS-NECs to be genetically distinct from neuroendocrine tumors (GIS-NET) in the same location. Clear genomic differences were also evident between pancreatic NECs (Panc-NEC) and nonpancreatic GIS-NECs (Nonpanc-NEC). Panc-NECs could be classified into two subgroups (i.e., "ductal-type" and "acinar-type") based on genomic features. Alterations in TP53 and RB1 proved common in GIS-NECs, and most Nonpanc-NECs with intact RB1 demonstrated mutually exclusive amplification of CCNE1 or MYC. Alterations of the Notch gene family were characteristic of Nonpanc-NECs. Transcription factors for neuroendocrine differentiation, especially the SOX2 gene, appeared overexpressed in most GIS-NECs due to hypermethylation of the promoter region. This first comprehensive study of genomic alterations in GIS-NECs uncovered several key biological processes underlying genesis of this very lethal form of cancer., Significance: GIS-NECs are genetically distinct from GIS-NETs. GIS-NECs arising in different organs show similar histopathologic features and share some genomic features, but considerable differences exist between Panc-NECs and Nonpanc-NECs. In addition, Panc-NECs could be classified into two subgroups (i.e., "ductal-type" and "acinar-type") based on genomic and epigenomic features. This article is highlighted in the In This Issue feature, p. 587., (©2021 The Authors; Published by the American Association for Cancer Research.)

Previous content analysis of video advertisements for functional foods identified "Exemption" advertisements. "Exemption" advertisements may imply to the audiences that "By taking functional foods, I can adopt unhealthy behaviors or I don't have to adopt healthy behaviors". In the context of Compensatory Health Beliefs (CHBs), this study refers to these beliefs as functional foods related to CHBs (FF-CHBs). This study aimed to assess the effects of exposure to "Exemption" advertisements for fat-reduction functional foods on audiences. The main hypothesis is exposure to "Exemption" video advertisements increases participants' FF-CHBs. Participants ( n   =  788) were randomly assigned to an intervention group that viewed three video advertisements or a control group and answered online self-administered questionnaires. Intervention videos were three videos with the highest number of views per month from among the "Exemption" video advertisements. Control videos were about how to brew green tea. FF-CHBs was assessed before and after the intervention. The intervention group showed significantly greater FF-CHBs after intervention (mean = 2.37 vs. 2.11, p < 0.001, η 2 = 0.026) compared with the control group. "Exemption" functional foods video advertisements increased FF-CHBs that can lead to adopting unhealthy behaviors and avoiding healthy behaviors. The content of these functional foods video advertisements should be improved to promote public health.

We evaluated the effects of a constant low (5-5%) and modulated (5-20%) oxygen environments on the in vitro development of bovine oocyte-cumulus-granulosa cell complexes (OCGCs) cultured in the presence or absence of an antioxidant (astaxanthin: Ax). OCGCs were cultured in a gas permeable culture device for 8 days in 5-5% O 2 (±Ax) and 5-20% O 2 (±Ax) culture conditions. In the oxygen modulated culture conditions, the oxygen concentration was switched from 5% to 20% on day 4 of culture. Ax promoted the viability of OCGCs (P < 0.05), but both oxygen and Ax had a significant effect on ROS production levels by OCGCs (P < 0.05). Specifically, ROS levels were significantly lower and higher under 5-5% O 2 (+Ax) and 5-20% O 2 (-Ax) conditions, respectively (P < 0.05), with intermediate levels observed in the 5-5% O 2 (-Ax) and the 5-20% O 2 (+Ax) culture conditions. The steroidogenic pattern was characterized by increasing estradiol-17β but with constant progesterone production levels regardless of culture conditions, suggesting the inhibition of luteinization-like changes in granulosa cells. OCGCs cultured in the 5-20% O 2 (+Ax) had higher nuclear maturation rates (P < 0.05) that were similar to the oocytes grown in vivo. However, there was no clear difference in the subsequent cleavage rates among the 5-5% O 2 (±Ax) and the 5-20% O 2 (+Ax) culture conditions (P > 0.05). A constant low oxygen environment significantly promoted the blastocyst rates (P < 0.05); however, the presence of Ax in the 5-20% O 2 (+Ax) condition also promoted development similar to the OCGCs cultured in the 5-5% O 2 (-Ax) condition (P > 0.05). In conclusion, exposure of OCGCs to constant low oxygen or oxygen modulation in the presence of Ax promotes the healthy development of OCGCs during the 8-day IVG culture using the gas permeable culture device., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Impaired oocyte quality is one of the main causes of low fertility in modern high-yielding dairy cows. One of the potential factors of the impaired oocyte quality is the effects of free fatty acids (FFA). In fact, high FFA supplementation to culture media exacerbated oocyte developmental competence in vitro. Meanwhile, artificially induced high blood FFA levels in heifers did not affect the lipid composition of oocytes in vivo; however, the oocyte lipid profile of postpartum cows has not yet been investigated. Therefore, the profile of lipids involved in energy metabolism, including FFA and triacylglycerols (TAG), and their relationship between plasma and oocytes were compared among cows at different lactation stages. Heifers were used as a control group that was not affected by lactation. Plasma and oocytes were collected from heifers (n = 4) and 14 Holstein cows categorized to the early lactation stage: 25-47 days in milk (DIM) (n = 6), peak lactation stage: 61-65 DIM (n = 4), and middle lactation stage: 160-202 DIM (n = 4). The FFA and TAG profiles of plasma and oocytes were examined by liquid chromatography mass spectrometry. Plasma FFA positively correlated with oocyte TAG (P < 0.05). Plasma FFA and oocyte TAG were significantly higher in cows in the early lactation stage than in heifers (P < 0.05), while the peak and middle lactation stage groups had intermediate levels. The proportion of oleic acid in plasma increased concurrently with elevations in total FFA, while the compositions of oocyte FFA and TAG fatty acyls were constant regardless of plasma FFA concentration or oocyte TAG content. The present results suggest that high postpartum plasma FFA concentrations affect the quantity of oocyte TAG. Taken together with the adverse effects of high FFA concentrations on oocyte developmental competence in vitro, oocyte quality in postpartum cows may be impaired due to high circulating FFA concentrations. These results provide a more detailed understanding of the effects of postpartum high circulating FFA concentrations on the low fertility of cows., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Blastocyst formation gives rise to the inner cell mass (ICM) and trophectoderm (TE) and is followed by the differentiation of the epiblast (Epi) and primitive endoderm (PrE) within the ICM. Although these two-round cell lineage differentiations underpin proper embryogenesis in every mammal, their spatiotemporal dynamics are quite diverse among species. Here, molecular details of the blastocyst stage in cattle were dissected using an optimized in vitro culture method. Blastocyst embryos were placed on agarose gel filled with nutrient-rich media to expose embryos to both gaseous and liquid phases. Embryos derived from this "on-gel" culture were transferred to surrogate mothers on day (D) 10 after fertilization and successfully implanted. Immunofluorescent studies using on-gel-cultured embryos revealed that the proportion of TE cells expressing the pluripotent ICM marker, OCT4, which was beyond 80% on D8, was rapidly reduced after D9 and reached 0% on D9.5. This first lineage segregation process was temporally parallel with the second one, identified by the spatial separation of Epi cells expressing SOX2 and PrE cells expressing SOX17. RNA-seq comparison of TE cells from D8 in vitro fertilized embryos and D14 in vivo embryos revealed that besides drastic reduction of pluripotency-related genes, TE cells highly expressed Wnt, FGF, and VEGF signaling pathways-related genes to facilitate the functional maturation required for feto-maternal interaction. Quantitative PCR analysis of TE cells derived from on-gel culture further confirmed time-dependent increments in the expression of key TE markers. Altogether, the present study provides platforms to understand species-specific strategies for mammalian preimplantation development., (© 2021 Federation of American Societies for Experimental Biology.)

Background: Intra-tumor heterogeneity (ITH) encompasses cellular differences in tumors and is related to clinical outcomes such as drug resistance. However, little is known about the dynamics of ITH, owing to the lack of time-series analysis at the single-cell level. Mouse models that recapitulate cancer development are useful for controlled serial time sampling., Results: We performed single-cell exome and transcriptome sequencing of 200 cells to investigate how ITH is generated in a mouse colorectal cancer model. In the model, a single normal intestinal cell is grown into organoids that mimic the intestinal crypt structure. Upon RNAi-mediated downregulation of a tumor suppressor gene APC, the transduced organoids were serially transplanted into mice to allow exposure to in vivo microenvironments, which play relevant roles in cancer development. The ITH of the transcriptome increased after the transplantation, while that of the exome decreased. Mutations generated during organoid culture did not greatly change at the bulk-cell level upon the transplantation. The RNA ITH increase was due to the emergence of new transcriptional subpopulations. In contrast to the initial cells expressing mesenchymal-marker genes, new subpopulations repressed these genes after the transplantation. Analyses of colorectal cancer data from The Cancer Genome Atlas revealed a high proportion of metastatic cases in human subjects with expression patterns similar to the new cell subpopulations in mouse. These results suggest that the birth of transcriptional subpopulations may be a key for adaptation to drastic micro-environmental changes when cancer cells have sufficient genetic alterations at later tumor stages., Conclusions: This study revealed an evolutionary dynamics of single-cell RNA and DNA heterogeneity in tumor progression, giving insights into the mesenchymal-epithelial transformation of tumor cells at metastasis in colorectal cancer., (© 2021. The Author(s).)

Cardiac responses to appetitive stimuli have been studied as indices of motivational states and attentional processes, the former being associated with cardiac acceleration and latter deceleration. Very few studies have examined heart rate changes in appetitive classical conditioning in humans. The current study describes the development and pilot testing of a classical conditioning task to assess cardiac responses to appetitive stimuli and cues that reliably precede them. Data from 18 adults were examined. They were shown initially neutral visual stimuli (putative CS) on a computer screen followed by pictures of high-caloric food (US). Phasic cardiac deceleration to food images was observed, consistent with an orienting response to motivationally significant stimuli. Similar responses were observed to non-appetitive stimuli when they were preceded by the cue associated with the food images, suggesting that attentional processes were engaged by conditioned stimuli. These autonomic changes provide significant information about classical conditioning effects in humans., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Sayão, Alves, Furukawa, Schultz Wenk, Cagy, Gutierrez-Arango, Tripp and Caparelli-Dáquer.)

Background: There is increasing evidence that the stochastic resonance (SR) phenomenon provided by subthreshold mechanical noise stimulation improves the sensory-motor system. However, the effect of SR on children with developmental coordination disorder (DCD) is unclear. The purpose of this study was to assess whether SR activated by subthreshold vibrotactile noise stimulation of the wrist influences manual dexterity in children with DCD. Methods: A double-blind interventional study was conducted. Participants were 30 children (age: 9.3 ± 1.44 years, range 6-11 years; 27 male, three female; 25 right-handed, five left-handed) meeting DCD diagnostic criteria in DSM-5. The manual dexterity test was administered the day before SR intervention (baseline-data). SR was elicited using subthreshold vibrotactile noise stimulation at 60% of the vibrotactile threshold measured at the wrist. SR was delivered two times and the manual dexterity test was administered during each SR stimulation block (SR-on condition) and after each SR stimulation block (SR-off), for a total of four measurements. Target outcomes were the component score, the standard score, and the percentile score of the manual dexterity test. Results: The manual dexterity test scores in the SR-on condition were significantly improved compared to scores at the baseline and in the SR-off condition ( p < 0.001). Conclusions: The present study showed that subthreshold noise stimulation eliciting SR significantly improved manual dexterity outcomes in children with DCD during stimulation but not after stimulation. Future studies will need to investigate the carry-over effects of SR stimulation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Nobusako, Osumi, Matsuo, Furukawa, Maeda, Shimada, Nakai and Morioka.)

Background: Previous studies have suggested that children with developmental coordination disorder (DCD) rely heavily on vision to perform movements, which may contribute to their clumsy movements. However, few studies have objectively and quantitatively investigated the perceptual biases of children with DCD., Methods: A visual-tactile temporal order judgment (TOJ) task was used to measure and compare the perceptual biases of 19 children with DCD and 19 age- and sex-matched typically developing children. The point of subjective equality, which demonstrates when "visual first" and "tactile first" judgment probabilities are equal (50%), obtained by analyzing the results of the visual-tactile TOJ task, was used as an indicator of perceptual biases. Further, variables (age and manual dexterity in all participants; motor function, autism spectrum disorder and attention-deficit hyperactivity disorder traits, and depressive symptoms in children with DCD) associated with perceptual biases were examined with correlation analysis., Results: Children with DCD had significantly stronger visual bias than typically developing children. Overall correlation analysis showed that increased visual bias was significantly correlated with poor manual dexterity., Conclusion: Children with DCD had a strong visual bias, which was associated with poor manual dexterity., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: There is increasing evidence that children with developmental coordination disorder (DCD) have deficits in sensory-motor integration, but it is unclear whether the sense of agency (SoA) generated by sensory-motor integration is altered., Aims: To investigate whether there is a difference in the time window for SoA between children with DCD and typically developing (TD) children., Methods and Procedures: An agency attribution task was used to quantitatively measure and compare the time window for SoA in 15 children with DCD and 46 children in the TD group. Variables that correlated with the time window for SoA were also examined in both groups of children., Outcomes and Results: The time window for SoA was significantly extended in children with DCD compared to TD children. The time window for SoA in TD children was significantly associated with manual dexterity, whereas the time window for SoA in children with DCD was significantly associated with depressive tendency., Conclusions and Implications: The time window for SoA is altered in children with DCD. The present results suggest that there may be a bidirectional relationship between an internal model deficit and depressive tendency and SoA in children with DCD., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

We report a patient with alexia with agraphia accompanied by letter-by-letter reading after hemorrhage in the left middle and inferior occipital gyri that spared the angular gyrus and the fusiform gyrus. Kanji (Japanese morphograms) and kana (Japanese phonetic writing) reading and writing tests revealed that alexia with agraphia was characterized by kana-predominant alexia and kanji-predominant agraphia. This type of "dorsal" letter-by-letter reading is discernable from conventional ventral type letter-by-letter reading that is observed in pure alexia in that (1) kinesthetic reading is less effective, (2) kana or literal agraphia coexists, and (3) fundamental visual discrimination is nearly normal.

Background: Approximately one-third of Japanese couples currently worry or previously worried about infertility. To develop strategies for the primary prevention of infertility as a population approach, it is important for the general population to be knowledgeable about fertility and infertility. The internet may contribute to the dissemination of information regarding infertility and fertility. However, few studies have examined online information about fertility., Objective: This study aimed to quantitatively examine online Japanese-language information about lifestyle factors associated with reduced fertility., Methods: We conducted online searches, using the 10 search terms with the highest numbers of searches that people hoping to conceive are likely to input in two major search engines in Japan (Google Japan and Yahoo! Japan). From the 2200 retrieved websites, 1181 duplicates and 500 websites unrelated to our objective were excluded, resulting in a final dataset of 519 websites. Coding guidelines were developed for the following lifestyle factors associated with reduced fertility: sexually transmitted diseases, psychological stress, cigarette smoking, alcohol use, nutrition and diet, physical activity and exercise, underweight, overweight and obesity, and environmental pollutants., Results: In terms of the website author's professional expertise, 69.6 % of the coding instances for the selected lifestyle factors were mentioned by hospitals, clinics, or the media, whereas only 1.7% were mentioned by laypersons. Psychological stress (20.1%) and sexually transmitted diseases (18.8%) were the most frequently mentioned lifestyle factors associated with reduced fertility. In contrast, cigarette smoking, alcohol use, nutrition and diet, physical activity and exercise, underweight, overweight and obesity, and environmental pollutants were mentioned relatively infrequently. The association between reduced fertility and sexually transmitted diseases was mentioned significantly more frequently by hospitals and clinics than by the media (P<.001). The association between reduced fertility and nutrition and diet was mentioned significantly more frequently by the media than by hospitals and clinics (P=.008). With regard to the sex of the target audience for the information, female-specific references to psychological stress, sexually transmitted diseases, nutrition and diet, underweight, physical activity and exercise, and overweight and obesity were significantly more frequent than were male-specific references to these lifestyle factors (psychological stress: P=.002, sexually transmitted diseases: P<.001, nutrition and diet: P<.001, underweight: P<.001, physical activity and exercise: P<.001, overweight and obesity: P<.001)., Conclusions: Of the lifestyle factors known to be related to reduced fertility, cigarette smoking, alcohol use, and male-specific lifestyle factors are mentioned relatively infrequently in online information sources in Japan, and these factors should be discussed more in information published on websites., (©Rie Yokota, Tsuyoshi Okuhara, Haruka Ueno, Hiroko Okada, Emi Furukawa, Takahiro Kiuchi. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 25.08.2020.)

A comparative lipidomic profiling analysis of dairy cattle oocytes with different developmental competences was performed using a combination of high performance liquid chromatography-high resolution tandem mass spectrometry and multivariate statistical analysis. Significant lipidomic changes were identified in degenerating oocytes. Total triacylglycerol in the degenerating oocytes was 1.8-fold higher than that in the normal oocytes; however, total cardiolipin was 53.5% lesser than that in the normal oocytes, which indicated attenuation of energy metabolism. Compared to those in the normal oocytes, triacylglycerols in the degenerating oocytes were composed of longer and more unsaturated acyl chains. In contrast, the acyl chains in free fatty acids present in the degenerating oocytes were shorter and with lesser degree of unsaturation compared to those in the normal oocytes. Moreover, a significant decrease in degenerating oocytes were found in total phosphatidylinositol (14.8 ± 7.6 pmol vs. 24.8 ± 5.5 pmol), total phosphatidylcholine (20.8 ± 8.7 pmol vs. 33.5 ± 7.2 pmol), and total plasmalogen ethanolamine (9.0 ± 4.7 pmol vs. 16.8 ± 5.2 pmol), which indicated dysfunction of lipid-metabolizing enzymes in oocytes during degeneration. Thus, increase of triacylglycerols together with the decrease of certain phospholipid species could be potential markers of oocyte developmental competence. In addition to providing a new approach to investigate the lipidomic changes in oocyte development, the lipidomic profiling in the present study has revealed insights that hold potential to unravel the role of lipid metabolism in oocyte developmental competence in cattle., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2019 Elsevier Inc. All rights reserved.)

There is growing recognition that much of human behavior is governed by the presence of classically conditioned cues. The Pavlovian-to-Instrumental Transfer (PIT) paradigm offers a way to measure the effects of classically conditioned stimuli on behavior. In the current study, a novel behavioral task, an adaptation of the PIT framework, was developed for use in conjunction with an fMRI classical conditioning task. Twenty-four healthy young adults completed (1) instrumental training, (2) Pavlovian conditioning, and (3) a Transfer test. During instrumental training, participants learned to apply force to a handgrip to win money from slot machines pictured on a computer screen. During Pavlovian conditioning, slot machines appeared with one of two abstract symbols (cues), one symbol was predictive of monetary reward. During the Transfer test, participants again applied force to a handgrip to win money. This time, the slot machines were presented with the Pavlovian cues, but with the outcomes hidden. The results indicated increased effort on the instrumental task, i.e. higher response frequency and greater force, in the presence of the reward-predicting cue. Our findings add to the growing number of studies demonstrating PIT effects in humans. This new paradigm is effective in measuring the effects of a conditioned stimulus on behavioral activation., (Copyright © 2020 da Costa, Furukawa, Hoefle, Moll, Tripp and Mattos.)

Although the media can have both negative and positive effects on children's cognitive and motor functions, its influence on their perceptual bias and manual dexterity is unclear. Thus, we investigated the association between media viewing time, media preference level, perceptual bias, and manual dexterity in 100 school-aged children. Questionnaires completed by children and their parents were used to ascertain media viewing time and preference levels. Perceptual bias and manual dexterity were measured using the visual-tactile temporal order judgment task and Movement Assessment Battery for Children-2nd edition, respectively. There were significant positive correlations between age and media viewing time and between media viewing time and media preference level. There was also a significant negative correlation between visual bias and manual dexterity. Hierarchical multiple regression analysis revealed that increasing visual bias was a significant predictor of decreasing manual dexterity. Further, children with low manual dexterity showed significant visual bias compared to those with high manual dexterity, when matched for age and gender. The present results demonstrated that, in school-aged children, although viewing media was not associated with perceptual bias and manual dexterity, there was a significant association between perceptual bias and manual dexterity.

Background: The timing of the first postpartum ovulation is an important factor affecting the timing of estrous resumption in dairy cows. The first postpartum ovulation is delayed in cows producing large amounts of milk with an intensive negative energy balance. The antral follicle count (AFC) and serum anti-Müllerian hormone concentrations are known to be indicators of the ovarian reserve, which is the number and quality of follicles left in a pair of ovaries and known as an indicator of female fertility. Cows with higher AFC have been proven to show higher pregnancy rate and shorter calving to conception intervals; however, the relationship between the timing of the first postpartum ovulation and ovarian reserve remains unclear. Therefore, this study examined the relationships between postpartum follicular dynamics, the ovarian cycle, nutritional status, and ovarian reserve., Methods: Transrectal ultrasonography was conducted from calving to 70-120 days in milk (DIM) in 26 cows to monitor AFC, follicular dynamics and the ovarian cycle. Body weight (BW) and milk yield were used as indicators of nutritional status., Results: The first postpartum ovulation was significantly later in cows with low AFC (< 25) than in those with high AFC (≥25), while changes in BW from calving to the nadir and milk production were similar in both groups. The present results also suggested that cows with low AFC and a delayed first postpartum ovulation had a shorter first ovarian cycle after the first postpartum ovulation. The mean DIM of the first postpartum artificial insemination (AI) and days open (days from calving to AI with which pregnancy was achieved) were similar in high and low AFC groups., Conclusions: The first postpartum ovulation was significantly earlier in cows with high AFC than in those with low AFC. The assumed reason for this result was higher sensitivity to luteinizing hormone and larger androstenedione and estradiol production in follicles in high AFC cows. Therefore, cows with high AFC may be more fertile than those with low AFC while their milk production increase and BW decrease; it means they are in negative energy balance. (340/350 words).

Attention deficit hyperactivity disorder (ADHD) has been associated with neural hyposensitivity to reward-predicting cues. Methylphenidate is widely used in the management of the disorder's symptoms, but its effects on reward sensitivity in ADHD are unknown. The current study used fMRI to measure striatal responses to reward-predicting cues in adults with ADHD on and off methylphenidate and a control group, during a classical conditioning task. Responses to cued reward were also explored. Larger differences in the ventral striatum activation to reward cues versus non-reward cues were observed when the ADHD participants were on methylphenidate compared to placebo. In response to cued-reward outcome, an exploratory analysis showed methylphenidate reduced the BOLD time-series correlation between the dorsal striatum and dorsal medial prefrontal cortex. Methylphenidate's therapeutic effects may be mediated by altering reward processing in individuals with ADHD., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Heat stress negatively affects reproductive functions in cows. Increased temperature disturbs fetal development in utero. However, the effect of heat stress on uterine endometrial tissues has not been fully examined. Using qPCR analysis, we measured the mRNA expression of various molecular markers in uterine endometrial tissue of dairy cows from Hokkaido, Japan, in winter and summer. Markers examined were heat shock proteins (HSPs), antioxidant enzymes (catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, and glutathione peroxidase 4), inflammatory cytokines, and interferon stimulated genes. Our results showed heat stress, body and milk temperatures were higher during summer than during winter. Expression levels of HSP27, HSP60, and HSP90 mRNA, and of catalase and copper/zinc superoxide dismutase mRNA were lower in summer than in winter. Tumor necrosis factor alpha expression was higher in summer than in winter. In conclusion, summer heat stress may reduce the expression of HSPs, affecting the levels of inflammatory cytokines in bovine uterine endometrial tissue., (© 2020 Japanese Society of Animal Science.)

Research on motivational processes in attention deficit/hyperactivity disorder (ADHD) focuses on reward. Studies with punishment are limited and findings mixed. This study evaluated the effects of punishment on response allocation in Japanese children with and without ADHD. Thirty-four children meeting DSM-IV criteria for ADHD and 59 typically developing control-group children completed an operant task in which they choose between playing two simultaneously available games. Reward was arranged symmetrically across the games under concurrent variable interval schedules. Asymmetric punishment schedules were superimposed with responses on one game punished four times as often as responses on the other. Children with ADHD showed greater behavioral sensitivity to punishment than controls. They allocated significantly more responses to the less frequently punished alternative and were more likely to play this game on consecutive trials and responded more slowly to the more punished game. Control group children allocated their responses evenly across games. Punishment exerted greater control over the behavior of Japanese children with ADHD than controls, similar to findings with children from Western countries, suggesting this is a common characteristic of the disorder. The behavior of typically developing Japanese children, while demonstrating awareness of punishment, was not controlled by the frequency of its occurrence.

Background: Transcatheter arterial chemoembolization (TACE) is widely used for unresectable hepatocellular carcinoma (HCC). The purpose of this study was to investigate incidence and risk factors of contrast-induced nephropathy (CIN) after TACE in patients with HCC., Methods: In this single-center retrospective study, we examined 461 consecutive TACE sessions in 260 patients between January 2003 and October 2015. CIN was defined as an increase in serum creatinine levels by ≥ 0.5 mg/dl or ≥ 25% from baseline within 72 h after TACE. We calculated incidence rate of CIN and tried to identify its risk factors by logistic regression analysis., Results: Twenty-one cases of CIN (5%) were observed in 461 TACE sessions. One patient required subsequent hemodialysis transiently. In univariate analysis, tumor size > 5 cm [odds ratio (OR) 5.76, 95% confidence interval (CI) 2.34-14.14, p < 0.001], chronic kidney disease (OR 2.54, 95% CI 1.05-6.14, p = 0.04), serum hemoglobin level [OR 0.79 (per 1 g/dl increase), 95% CI 0.64-0.98, p = 0.03] and serum albumin level [OR 0.44 (per 1 g/dl increase), 95% CI 0.19-1.02, p = 0.05] were associated with the development of CIN. Stepwise logistic regression methods showed that tumor size > 5 cm (OR 7.81, 95% CI 2.99-20.46, p < 0.001) and serum albumin [OR 0.29 (per 1 g/dl increase), 95% CI 0.11-0.75, p = 0.01] were risk factors of CIN., Conclusions: In this study, HCC tumor size and lower serum albumin level were independent predictors of CIN after TACE.

Altered reinforcement sensitivity is hypothesized to underlie symptoms of attention deficit hyperactivity disorder (ADHD). Here we evaluate the behavioral sensitivity of Brazilian children with and without ADHD to a change in reward availability. Forty typically developing children and 32 diagnosed with DSM-IV ADHD completed a signal-detection task in which correct discriminations between two stimuli were associated with different frequencies of reinforcement. The response alternative associated with the higher rate of reinforcement switched, without warning, after 30 rewards were delivered. The task continued until another 30 rewards were delivered. Both groups of children developed a response bias toward the initially more frequently reinforced alternative. This effect was larger in the control group. The response allocation of the two groups changed following the shift in reward availability. Over time the ADHD group developed a significant response bias toward the now more frequently reinforced alternative. In contrast, the bias of the control group stayed near zero after an initial decline following the contingency change. The overall shift in bias was similar for the two groups. The behavior of both groups of children was sensitive to the asymmetric reward distribution and to the change in reward availability. Subtle group differences in response patterns emerged, possibly reflecting differences in the time frame of reward effects and sensitivity to reward exposure.

Next-generation sequencing (NGS) of tumor tissue (ie, clinical sequencing) can guide clinical management by providing information about actionable gene aberrations that have diagnostic and therapeutic significance. Here, we undertook a hospital-based prospective study (TOP-GEAR project, 2nd stage) to investigate the feasibility and utility of NGS-based analysis of 114 cancer-associated genes (the NCC Oncopanel test). We examined 230 cases (comprising more than 30 tumor types) of advanced solid tumors, all of which were matched with nontumor samples. Gene profiling data were obtained for 187 cases (81.3%), 111 (59.4%) of which harbored actionable gene aberrations according to the Clinical Practice Guidelines for Next Generation Sequencing in Cancer Diagnosis and Treatment (Edition 1.0) issued by 3 major Japanese cancer-related societies. Twenty-five (13.3%) cases have since received molecular-targeted therapy according to their gene aberrations. These results indicate the utility of tumor-profiling multiplex gene panel testing in a clinical setting in Japan. This study is registered with UMIN Clinical Trials Registry (UMIN 000011141)., (© 2019 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Objectives: Trimethoprim/sulfamethoxazole (SXT) is used to treat Staphylococcus aureus infections. However, the effect of treatment with SXT is sometimes not sufficient and there are patients whose treatment has to be prolonged. There are few reports of isolated strains of SXT-resistant S. aureus, but it is possible that some resistant strains cannot be detected by current testing methods We have therefore developed a tool to identify these resistant strains., Methods: The mutant selection window (MSW) of SXT for 40 clinical isolates of S. aureus, including 20 methicillin-resistant S. aureus (MRSA), was determined. The optimum concentration of SXT and thymidine in agar for detecting SXT-resistant small colony variants (SCVs) of S. aureus was investigated., Results: The MSW 50 and MSW 90 of SXT, presented as a multiple of the minimum inhibitory concentration (MIC), were 16× MIC and >256× MIC, respectively. SCVs were detected within the MSW in 32 (80%) of the 40 clinical isolates studied. To maintain the morphology of SCVs, the most suitable concentrations of SXT and thymidine for screening were 4mg/L and 0.01μg/mL, respectively. All 32 SCVs were resistant to SXT (MIC >32mg/L). The sensitivity and specificity of this screening method was 100% and 88.9%, respectively., Conclusions: SXT-resistant SCVs are not usually detected by routine laboratory tests performed in hospitals. However, the screening test described here can easily distinguish SXT-resistant SCVs among S. aureus isolated from specimens. This newly developed screening test could become an important tool to prevent inappropriate use of SXT., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Eicosapentaenoic acid (EPA)/arachidonic acid (AA) ratio showed inverse associations with cardiovascular disease (CVD) in general population. However, this has not been examined enough in dialysis patients. We cross-sectionally investigated the relationship between EPA/AA ratio and prevalence of CVD in 321 chronic hemodialysis patients (64 ± 11 years old; 110 women; dialysis vintage 10 ± 8 years) in an urban area of Tokyo. CVD was defined as a composite of ischemic heart disease, ischemic stroke and hemorrhagic stroke. The frequency of dietary fish intake was also examined. Logistic regression was used to quantify the association of EPA/AA ratio with CVD. EPA/AA ratio was 0.31 ± 0.19 and 154 patients (48%) consumed fish once or less weekly. One hundred and thirty patients (41%) had CVD, including 65 with ischemic heart disease, 70 with ischemic stroke, and 20 with hemorrhagic stroke. Age (odds ratio [OR], 1.04; P = 0.01), hypertension (OR, 2.25; P = 0.002), and dialysis vintage (OR, 1.04; P = 0.02) were associated with CVD; however, EPA/AA was not after adjustment for other risk factors. A similar relationship was observed between fish intake and CVD prevalence. We did not find any significant association between EPA/AA ratio and prevalence of CVD, although traditional risk factors such as age, hypertension and dialysis vintage were associated with CVD. These results might have been influenced by the fact that only a small proportion of our patients showed a high EPA/AA ratio., (© 2018 International Society for Apheresis, Japanese Society for Apheresis, and Japanese Society for Dialysis Therapy.)

Weight reduction is important in patients with sleep-disordered breathing (SDB). In Japanese patients, slight weight reduction is effective for improving the severity of SDB. However, the effect of weight reduction after administration of sodium glucose co-transporter 2 (SGLT2) inhibitor for SDB remains unclear. The aim of this study was to evaluate the improvement of SDB from baseline after administration of dapagliflozin (5 mg) once daily for 24 weeks among Japanese patients with obesity and type 2 diabetes mellitus. Thirty Japanese patients with type 2 diabetes mellitus and SDB were enrolled in a 24-week, prospective, open-label, single-arm, multicentre trial. SDB was defined as at least five 3% oxygen desaturation index (ODI) events per hour, and moderate to severe SDB was defined as at least 15 ODI events per hour. The primary endpoint was the change in 3% ODI between before dapagliflozin administration and at 24 weeks. The prevalence of moderate to severe SDB was 20% in the present study. After administration of dapagliflozin, fasting glucose, HbA1c, aspartate aminotransferase, total cholesterol, low-density lipoprotein cholesterol, and estimated globular filtration rate decreased significantly. The improvement of 3% ODI was observed in patients with moderate to severe SDB but not mild SDB (from 25.0 ± 3.8 at baseline to 18.5 ± 6.1 at 24 weeks, p = 0.017). In conclusion, dapagliflozin might improve moderate to severe SDB but not mild SDB in Japanese patients with obesity and type 2 diabetes mellitus.

Transglutaminase (TG) is an essential enzyme to catalyze cross-linking reactions of epidermal proteins. Recently, we biochemically characterized human skin TG orthologues for medaka (Oryzias latipes), a model fish. By genome editing, gene-modified fishes for the two orthologues were obtained, both of which lack the ordinal enzymes. These fish appeared to exhibit higher susceptibility to osmolality at the period of larvae.

Although visuo-motor temporal integration in children is suggested to be related to motor control and motor learning, its relevance is still unclear. On the other hand, visuo-motor temporal integration ability undergoes developmental changes with age. In the current correlational study, we measured manual dexterity and visuo-motor temporal integration ability in 132 children with typical development (age, 4-15 years) and investigated the relationship between the two functions. The Movement Assessment Battery for Children-2nd edition was used as an indicator of manual dexterity. The delay detection threshold (DDT) and steepness of the probability curve for delay detection, which was measured by the delayed visual feedback detection task for self-generated movement, were used as indices of the visuo-motor temporal integration ability. The results indicated significant correlations between manual dexterity/age and DDT/steepness of the probability curve for delay detection. In addition, hierarchical multiple regression analysis showed that both manual dexterity and age significantly contributed to visuo-motor temporal integration, indicating a better fit than when only age was employed as an independent variable. Importantly, there was no interaction effect between age and manual dexterity. These findings were the first to suggest that manual dexterity is a significant predictor of visuo-motor temporal integration ability in children, regardless of age. The present study validated the important relationship between visuo-motor temporal integration and manual dexterity in children. Considering the limitations of the current study, including the non-homogeneous sample, further studies are still warranted to validate the results.

Advanced cancer genomics technologies are now being employed in clinical sequencing, where next-generation sequencers are used to simultaneously identify multiple types of DNA alterations for prescription of molecularly targeted drugs. However, no computational tool is available to accurately detect DNA alterations in formalin-fixed paraffin-embedded (FFPE) samples commonly used in hospitals. Here, we developed a computational tool tailored to the detection of single nucleotide variations, indels, fusions, and copy number alterations in FFPE samples. Elaborated multilayer noise filters reduced the inherent noise while maintaining high sensitivity, as evaluated in tumor-unmatched normal samples using orthogonal technologies. This tool, cisCall, should facilitate clinical sequencing in everyday diagnostics. It is available at https://www.ciscall.org .

The neurological basis of developmental coordination disorder (DCD) is thought to be deficits in the internal model and mirror-neuron system (MNS) in the parietal lobe and cerebellum. However, it is not clear if the visuo-motor temporal integration in the internal model and automatic-imitation function in the MNS differs between children with DCD and those with typical development (TD). The current study aimed to investigate these differences. Using the manual dexterity test of the Movement Assessment Battery for Children (second edition), the participants were either assigned to the probable DCD (pDCD) group or TD group. The former was comprised of 29 children with clumsy manual dexterity, while the latter consisted of 42 children with normal manual dexterity. Visuo-motor temporal integration ability and automatic-imitation function were measured using the delayed visual feedback detection task and motor interference task, respectively. Further, the current study investigated whether autism-spectrum disorder (ASD) traits, attention-deficit hyperactivity disorder (ADHD) traits, and depressive symptoms differed among the two groups, since these symptoms are frequent comorbidities of DCD. In addition, correlation and multiple regression analyses were performed to extract factors affecting clumsy manual dexterity. In the results, the delay-detection threshold (DDT) and steepness of the delay-detection probability curve, which indicated visuo-motor temporal integration ability, were significantly prolonged and decreased, respectively, in children with pDCD. The interference effect, which indicated automatic-imitation function, was also significantly reduced in this group. These results highlighted that children with clumsy manual dexterity have deficits in visuo-motor temporal integration and automatic-imitation function. There was a significant correlation between manual dexterity, and measures of visuo-motor temporal integration, and ASD traits and ADHD traits and ASD. Multiple regression analysis revealed that the DDT, which indicated visuo-motor temporal integration, was the greatest predictor of poor manual dexterity. The current results supported and provided further evidence for the internal model deficit hypothesis. Further, they suggested a neurorehabilitation technique that improved visuo-motor temporal integration could be therapeutically effective for children with DCD.

Community-acquired pneumonia and otitis media are caused by several bacterial species, including Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. For the treatment of these diseases, various quinolones are frequently used. We determined the mutant prevention concentration (MPC) of four quinolones, levofloxacin, sitafloxacin, tosufloxacin, and garenoxacin, using 92 clinical isolates and evaluated each mutant selection window (MSW). Furthermore, the DNA sequence of the quinolone resistance-determining region (QRDR) for the resistant mutant selected based on the MSW was determined. The MIC 90 and MPC 90 of levofloxacin were 0.781 μg/mL and 6.250 μg/mL for S. pneumoniae and 0.049 μg/mL and 1.563 μg/mL for M. catarrhalis and were higher than those for the other three quinolones. In addition, 5 strains of 30 S. pneumoniae (16.7%) selected based on the MSW of levofloxacin acquired resistance to only levofloxacin. In these 5 strains, a mutation of gyrA and/or parC was detected. In this study, no resistant mutant was selected in the MSW of any of the other three quinolones. On the other hand, clinical isolates of H. influenzae showed no resistance by all quinolone exposure. Finally, The MIC value and the mutation status in the QRDR did not change after 14 passages in antibiotic-free medium. In conclusion, our findings suggest that the increased use of levofloxacin may contribute to the increased quinolone-resistance of S. pneumoniae and M. catarrhalis., (Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: Most research on motivational processes in attention deficit hyperactivity disorder (ADHD) has been undertaken in Western Europe and North America. The extent to which these findings apply to other cultural groups is unclear. The current study evaluated the behavioral sensitivity of Japanese children with and without ADHD to changing reward availability. Forty-one school-aged children, 19 diagnosed with DSM-IV ADHD, completed a signal-detection task in which correct discriminations between two stimuli were associated with different reinforcement frequencies. The response alternative associated with the higher rate of reinforcement switched twice during the task without warning., Findings: Both groups of children developed an initial bias toward the more frequently reinforced response alternative. When the reward contingencies switched the response allocation (bias) of the control group children followed suit. The response bias scores of the children with ADHD did not, suggesting impaired tracking of reward availability over time., Conclusions: Japanese children with ADHD adjust their behavioral responses to changing reinforcer availability less than their typically developing peers. This is not explained by poor attention to task or a lack of sensitivity to reward. The current results are consistent with altered sensitivity to changing reward contingencies identified in non-Japanese samples of children with ADHD. Irrespective of their country of origin, children with ADHD will likely benefit from behavioral expectations and reinforcement contingencies being made explicit together with high rates of reinforcement for appropriate behaviors.

Lynch syndrome is a cancer predisposition syndrome caused by germline mutations in mismatch repair (MMR) genes. MMR deficiency is a ubiquitous feature of Lynch syndrome-associated colorectal adenocarcinomas; however, it remains unclear when the MMR-deficient phenotype is acquired during tumorigenesis. To probe this issue, the present study examined genetic alterations and MMR statuses in Lynch syndrome-associated colorectal adenomas and adenocarcinomas, in comparison with sporadic adenomas. Among the Lynch syndrome-associated colorectal tumors, 68 of 86 adenomas (79%) and all adenocarcinomas were MMR-deficient, whereas all the sporadic adenomas were MMR-proficient, as determined by microsatellite instability testing and immunohistochemistry for MMR proteins. Sequencing analyses identified APC or CTNNB1 mutations in the majority of sporadic adenomas (58/84, 69%) and MMR-proficient Lynch syndrome-associated adenomas (13/18, 72%). However, MMR-deficient Lynch syndrome-associated adenomas had less APC or CTNNB1 mutations (25/68, 37%) and frequent frameshift RNF43 mutations involving mononucleotide repeats (45/68, 66%). Furthermore, frameshift mutations affecting repeat sequences constituted 14 of 26 APC mutations (54%) in MMR-deficient adenomas whereas these frameshift mutations were rare in MMR-proficient adenomas in patients with Lynch syndrome (1/12, 8%) and in sporadic adenomas (3/52, 6%). Lynch syndrome-associated adenocarcinomas exhibited mutation profiles similar to those of MMR-deficient adenomas. Considering that WNT pathway activation sufficiently drives colorectal adenoma formation, the distinct mutation profiles of WNT pathway genes in Lynch syndrome-associated adenomas suggest that MMR deficiency commonly precedes adenoma formation.

Study Question: Can supplementation of medium with prolactin (PRL), epidermal growth factor (EGF) and 4-hydroxyestradiol (4-OH-E2) prior to embryo transfer improve implantation potential in mouse blastocysts derived from IVF?, Summary Answer: Combined treatment with PRL, EGF and 4-OH-E2 improves mouse blastocyst implantation rates, while alone, each factor is ineffective., What Is Known Already: Blastocyst dormancy during delayed implantation caused by ovariectomy is maintained by continued progesterone treatment in mice, and estrogen injection rapidly activates blastocysts to implantation-induced status in vivo. While the expression of many proteins is upregulated in implantation-induced blastocysts, selective proteolysis by proteasomes, such as estrogen receptor α (ESR1), occurs in implantation-induced blastocysts to achieve implantation-competent status. It is worth evaluating the proteins expressed during these periods to identify humoral factors that might improve the implantation potential of IVF-derived blastocysts because the poor quality of embryos obtained by IVF is one of the major causes of implantation failure., Study Design, Size, Duration: Superovulated oocytes from ICR mice were fertilized with spermatozoa and then cultured in vitro in potassium simplex optimized medium (KSOM) without phenol red (KSOM-P) for 90-96 h. Blastocysts were treated with PRL (10 or 20 mIU/mL), EGF (5 or 10 ng/mL) or 4-OH-E2 (1 or 10 nM) in KSOM-P for 24 h., Participants/materials, Setting, Methods: Levels of breast cancer 1 (BRCA1), EGF receptor (EGFR, also known as ERBB1), ERBB4, tubulointerstitial nephritis antigen-like 1 (TINAGL1) and ESR1 protein were examined with immunohistochemical analysis using immunofluorescence methods and confocal laser scanning microscopy. For embryo transfer, six blastocysts were suspended in HEPES-buffered KSOM-P medium and transferred into the uteri of recipient mice on the morning of Day 4 (0900-1000 h) of pseudopregnancy (Day 1 = vaginal plug). The number of implantation sites was then recorded on Day 6 using the blue dye method., Main Results and the Role of Chance: PRL, EGF and 4-OH-E2 each promoted BRCA1 protein level in the trophectoderm (TE). While PRL treatment resulted in an increase in EGFR, EGF increased both EGFR and ERBB4 in the blastocyst TE. TINAGL1 in the TE was enhanced by 4-OH-E2, which also increased localization of this protein to the basement membrane. Treatment with PRL, EGF or 4-OH-E2 alone did not improve blastocyst implantation rates. Combined treatment with PRL, EGF and 4-OH-E2 resulted in increased levels of EGFR, ERBB4, TINAGL1 and BRCA1 in the TE, whereas ESR1 was not upregulated in the treated blastocysts. Furthermore, combined treatment with PRL, EGF and 4-OH-E2 improved blastocyst implantation rates versus control (P = 0.009)., Large Scale Data: Not applicable., Limitations, Reasons for Caution: Our studies were carried out in a mouse model, and the conclusions were drawn from limited results obtained from one species. Whether the increase in EGFR, ERBB4 and TINAGL1 protein in the TE improves implantation potential of blastocysts needs to be further studied experimentally by assessing other expressed proteins. The influence of combined supplementation in vitro of PRL, EGF and 4-OH-E2 on implantation also requires further examination and optimization in human blastocysts before it can be considered for clinical use in ART., Wider Implications of the Findings: Enhanced implantation potential by combined treatment with PRL, EGF and 4-OH-E2 appears to result in the upregulation of at least two distinct mechanisms, namely signaling via EGF receptors and basement membrane formation during the peri-implantation period in mice. While PRL, EGF and 4-OH-E2 each promoted BRCA1 protein level in the TE, treatment with each alone did not improve blastocyst implantation. Therefore, BRCA1 protein appears to be unnecessary for the attachment reaction in blastocysts in mice Combined supplementation of PRL, EGF and 4-OH-E2 might also be of relevance for embryo transfer of human IVF-derived blastocysts for ART., Study Funding/competing Interest(s): This work was supported in part by the JSPS KAKENHI [Grant numbers 22580316 and 25450390 (to H.M.)] and the Joint Research Project of Japan-U.S. Cooperative Science Program (to H.M.). The authors have no conflict of interest to declare., (© The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com)

Gastrointestinal stromal tumors represent the most common mesenchymal tumor of the digestive tract, driven by gain-of-function mutations in KIT. Despite its proven benefits, half of the patients treated with imatinib show disease progression within 2 years due to secondary resistance mutations in KIT. It remains unclear how the genomic and transcriptomic features change during the acquisition of imatinib resistance. Here, we performed exome sequencing and microarray transcription analysis for four imatinib-resistant cell lines and one cell line briefly exposed to imatinib. We also performed exome sequencing of clinical tumor samples. The cell line briefly exposed to imatinib exhibited few single-nucleotide variants and copy-number alterations, but showed marked upregulation of genes related to detoxification and downregulation of genes involved in cell cycle progression. Meanwhile, resistant cell lines harbored numerous genomic changes: amplified genes related to detoxification and deleted genes with cyclin-dependent kinase activity. Some variants in the resistant samples were traced back to the drug-sensitive samples, indicating the presence of ancestral subpopulations. The subpopulations carried variants associated with cell death. Pre-existing cancer cells with genetic alterations promoting apoptosis resistance may serve as a basis whereby cancer cells with critical mutations, such as secondary KIT mutations, can establish full imatinib resistance. © 2017 The Authors Genes, Chromosomes and Cancer Published by Wiley Periodicals, Inc., (© 2017 The Authors Genes, Chromosomes and Cancer Published by Wiley Periodicals, Inc.)

Background: The behavioral sensitivity of children with ADHD to punishment has received limited theoretical and experimental attention. This study evaluated the effects of punishment on the response allocation of children with ADHD and typically developing children., Method: Two hundred and ten children, 145 diagnosed with ADHD, completed an operant task in which they chose between playing two simultaneously available games. Reward was arranged symmetrically across the games under concurrent variable interval schedules. Asymmetric punishment schedules were superimposed; responses on one game were punished four times as often as responses on the other., Results: Both groups allocated more of their responses to the less frequently punished alternative. Response bias increased significantly in the ADHD group during later trials, resulting in missed reward trials and reduced earnings., Conclusions: Punishment exerted greater control over the response allocation of children with ADHD with increased time on task. Children with ADHD appear more sensitive to the cumulative effects of punishment than typically developing children., (© 2016 Association for Child and Adolescent Mental Health.)

Nucleation, the primary step in crystallization, dictates the number of crystals, the distribution of their sizes, the polymorph selection, and other crucial properties of the crystal population. We used time-resolved liquid-cell transmission electron microscopy (TEM) to perform an in situ examination of the nucleation of lysozyme crystals. Our TEM images revealed that mesoscopic clusters, which are similar to those previously assumed to consist of a dense liquid and serve as nucleation precursors, are actually amorphous solid particles (ASPs) and act only as heterogeneous nucleation sites. Crystalline phases never form inside them. We demonstrate that a crystal appears within a noncrystalline particle assembling lysozyme on an ASP or a container wall, highlighting the role of heterogeneous nucleation. These findings represent a significant departure from the existing formulation of the two-step nucleation mechanism while reaffirming the role of noncrystalline particles. The insights gained may have significant implications in areas that rely on the production of protein crystals, such as structural biology, pharmacy, and biophysics, and for the fundamental understanding of crystallization mechanisms.

Protein induced by vitamin K absence or antagonist-II (PIVKA-II) is an abnormal prothrombin lacking gamma-carboxylation of the 10 glutamic-acid residues in the N-terminal region. PIVKA-II has been used as an effective biomarker of hepatocellular carcinoma (HCC) since the PIVKA-II level is not correlated with that of alpha-fetoprotein (AFP), which is another effective biomarker for HCC. Monoclonal antibodies for the clinical biomarker test are usually expensive because of their high production costs. Recently, many studies involving the expression of the recombinant globulin molecules in bacterial cells and plants have been con- ducted. These studies have enabled us to produce recombinant monoclonal antibodies at much lower costs. In this study, we first produced a hybridoma expressing a monoclonal antibody against PIVKA-II, and then we constructed and produced a single-chain fragment-variable antibody (scFv), created by the linking of variable regions of light- and heavy-chains of the PIVKA-II monoclonal antibody with a peptide linker of triplicated GGGGS. The scFv was expressed in E. coli and exhibited high specificity for PIVYKA-II binding, while its binding titer was low. [Review].

Mucinous gastric carcinoma (MGC) is a unique subtype of gastric cancer with a poor survival outcome. Comprehensive molecular profiles and putative therapeutic targets of MGC remain undetermined. We subjected 16 tumour-normal tissue pairs to whole-exome sequencing (WES) and an expanded set of 52 tumour-normal tissue pairs to subsequent targeted sequencing. The latter focused on 114 genes identified by WES. Twenty-two histologically differentiated MGCs (D-MGCs) and 46 undifferentiated MGCs (U-MGCs) were analysed. Chromatin modifier genes, including ARID1A (21%), MLL2 (19%), MLL3 (15%), and KDM6A (7%), were frequently mutated (47%) in MGC. We also identified mutations in potential therapeutic target genes, including MTOR (9%), BRCA2 (9%), BRCA1 (7%), and ERBB3 (6%). RHOA mutation was detected only in 4% of U-MGCs and in no D-MGCs. MYH9 was recurrently (13%) mutated in MGC, with all these being of the U-MGC subtype (p = 0.023). Three U-MGCs harboured MYH9 nonsense mutations. MYH9 knockdown enhanced cell migration and induced intracytoplasmic mucin and cellular elongation. BCOR mutation was associated with improved survival. In U-MGCs, the MLH1 expression status and combined mutation status (TP53/BCL11B or TP53/MLL2) were prognostic factors. A comparative analysis of driver genes revealed that the mutation profile of D-MGC was similar to that of intestinal-type gastric cancer, whereas U-MGC was a distinct entity, harbouring a different mutational profile to intestinal- and diffuse-type gastric cancers. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd., (Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)

Background: Altered sensitivity to positive reinforcement has been hypothesized to contribute to the symptoms of attention-deficit hyperactivity disorder (ADHD). In this study, we evaluated the ability of children with and without ADHD to adapt their behavior to changing reinforcer availability., Method: Of one hundred sixty-seven children, 97 diagnosed with ADHD completed a signal-detection task in which correct discriminations between two stimuli were associated with different frequencies of reinforcement. The response alternative associated with the higher rate of reinforcement switched twice during the task without warning. For a subset of participants, this was followed by trials for which no reinforcement was delivered, irrespective of performance., Results: Children in both groups developed an initial bias toward the more frequently reinforced response alternative. When the response alternative associated with the higher rate of reinforcement switched, the children's response allocation (bias) followed suit, but this effect was significantly smaller for children with ADHD. When reinforcement was discontinued, only children in the control group modified their response pattern., Conclusions: Children with ADHD adjust their behavioral responses to changing reinforcer availability less than typically developing children, when reinforcement is intermittent and the association between an action and its consequences is uncertain. This may explain the difficulty children with ADHD have adapting their behavior to new situations, with different reinforcement contingencies, in daily life., (© 2016 Association for Child and Adolescent Mental Health.)

Brain development requires a fine-tuned copper homoeostasis. Copper deficiency or excess results in severe neuro-pathologies. We demonstrate that upon neuronal differentiation, cellular demand for copper increases, especially within the secretory pathway. Copper flow to this compartment is facilitated through transcriptional and metabolic regulation. Quantitative real-time imaging revealed a gradual change in the oxidation state of cytosolic glutathione upon neuronal differentiation. Transition from a broad range of redox states to a uniformly reducing cytosol facilitates reduction of the copper chaperone Atox1, liberating its metal-binding site. Concomitantly, expression of Atox1 and its partner, a copper transporter ATP7A, is upregulated. These events produce a higher flux of copper through the secretory pathway that balances copper in the cytosol and increases supply of the cofactor to copper-dependent enzymes, expression of which is elevated in differentiated neurons. Direct link between glutathione oxidation and copper compartmentalization allows for rapid metabolic adjustments essential for normal neuronal function.