Fatty acid composition of the methylotrophic yeast Komagataella phaffii grown under low- and high-methanol conditions.

In this study, we analysed the intracellular fatty acid profiles of Komagataella phaffii during methylotrophic growth. K. phaffii grown on methanol had significantly lower total fatty acid contents in the cells compared with glucose-grown cells. C18 and C16 fatty acids were the predominant fatty acids in K. phaffii, although the contents of odd-chain fatty acids such as C17 fatty acids were also relatively high. Moreover, the intracellular fatty acid composition of K. phaffii changed in response to not only carbon sources but also methanol concentrations: C17 fatty acids and C18:2 content increased significantly as methanol concentration increased, whereas C18:1 and C18:3 contents were significantly lower in methanol-grown cells. The intracellular content of unidentified compounds (Cn H2n O4 ), on the other hand, was significantly greater in cells grown on methanol. As the intracellular contents of these Cn H2n O4 compounds were significantly higher in a gene-disrupted strain for glutathione peroxidase (gpx1Δ) than in the wild-type strain, we presume that the Cn H2n O4 compounds are fatty acid peroxides. These results indicate that K. phaffii can coordinate intracellular fatty acid composition during methylotrophic growth in order to adapt to high-methanol conditions and that certain fatty acid species such as C17:0, C17:1, C17:2 and C18:2 may be related to the physiological functions by which K. phaffii adapts to high-methanol conditions.

Baseline Body Composition and Physical Activity Level Recommended for Optimal Bone Mineral Density in Young Women.

Aim: This study examined the influence of body composition and physical activity level (PAL) on bone mineral density (BMD) to determine the baseline values necessary for maintaining healthy trabecular bone and improving bone health in young women. Materials and Methods: The subjects, consisting of healthy young women, were assigned to either a BMD-low (BMD-L) or BMD-high (BMD-H) group using the osteosonic index. PAL was measured for 1 week and rated on a scale from PAL-1 to 8 according to intensity levels (metabolic equivalents [METs]). The Wilcoxon rank-sum test was performed for intergroup comparisons. Results: The BMD-H group had significantly higher fat-free muscle mass, skeletal muscle index, and basic metabolic rate than the BMD-L group (p < 0.001, each). Regarding the timing of physical activity in daily life according to intensity, PAL-6 (p < 0.01), PAL-7 (p < 0.001), and PAL-8 (p < 0.01) for the BMD-H group were significantly longer than those for the BMD-L group. Discussion and Conclusion: For young women in their early 20s, BMD may be associated with baseline physical fitness and strength, as determined by body composition, but it is not influenced by the duration of physical activity. A PAL ≥6.0 METs may improve or maintain the effect on BMD.

Establishment of three malignant peripheral nerve sheath tumor cell lines, FU-SFT8611, 8710 and 9817: conventional and molecular cytogenetic characterization.

Malignant peripheral nerve sheath tumor (MPNST) is a rare malignant tumor, for which only a few cultured cell lines are available to date. In the present study, we established three new MPNST cell lines, FU-SFT8611, FU-SFT8710 and FU-SFT9817, from a 40-year-old Japanese man without neurofibromatosis 1 (NF1), a 43-year-old Japanese woman with NF1, and a 61-year-old Japanese woman without NF1, respectively. These cell lines were in culture for more than 3 years in vitro, and exhibit complex karyotypes lacking a common characteristic pattern. Two of the cell lines, FU-SFT8611 and FU-SFT9817, were tumorigenic in nude mice, with the resultant tumors showing an immunohistochemical phenotype similar to the original tumors. Comparative genomic hybridization analysis revealed that chromosomal imbalances were very similar between the original tumors and the established cell lines, but no consistent imbalances among the three cell lines as reported so far. As for chromosomal imbalances that have been reported to be associated with poor survival, FU-SFT8611 exhibited a gain of chromosome 17q22-qter, and FU-SFT9817 showed a loss of 17q22-qter, while gains of 17q24 and 7p15-p21 were observed in none of the cell lines. These newly established cell lines provide a valuable resource for biological and pathological investigations into new treatment regimes for MPNST.

Establishment of a new human synovial sarcoma cell line, FU-SY-1, that expresses c-Met receptor and its ligand hepatocyte growth factor.

Only a small number of human synovial sarcoma cell lines have been reported, and of those, not all have been fully characterized, especially at the molecular level. We describe here the establishment and characterization of a new human cell line, FU-SY-1, which originated from a monophasic fibrous synovial sarcoma arising in the supinator muscle of a 31-year-old woman. This cell line propagated continuously in vitro for 73 serial passages for more than 36 months. FU-SY-1 cells in vitro were rather small, exhibited a spindle or polygonal shape without conspicuous pleomorphism, and expressed c-Met and hepatocyte growth factor (HGF) as determined by immunocytochemistry. Cytogenetically, FU-SY-1 cells maintained a consistent karyotype: 47, X, +7, t(X;18)(p11.2;q11.2), the same as that of the original tumor specimen. Reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated a SYT-SSX fusion transcript and expression of c-Met and HGF mRNA in FU-SY-1 cells. A subsequent sequence analysis using the PCR products confirmed that the detected messages were derived from the SYT-SSX1 fusion gene. This cell line, FU-SY-1, established from a monophasic fibrous synovial sarcoma, may therefore be a useful tool for investigation of the mechanisms of tumorigenesis and progression in human synovial sarcomas.

Establishment of a novel human dedifferentiated liposarcoma cell line, FU-DDLS-1: conventional and molecular cytogenetic characterization.

A number of human cell lines derived from well-differentiated, myxoid/round cell, or pleomorphic liposarcoma have been described. To our knowledge, however, no human cell line established from dedifferentiated liposarcoma has been reported. In this study, we established a new human cell line, FU-DDLS-1, which originated from a dedifferentiated liposarcoma arising in the retroperitoneum of a 61-year-old man. This cell line was characterized by immunocytochemistry, conventional banding analysis, fluorescence in situ hybridization with chromosome painting probe, and comparative genomic hybridization (CGH). FU-DDLS-1 cells were spindle or polygonal shaped and possessed oval nuclei and slender cytoplasmic processes. The cultured cells were successfully maintained in vitro for over 90 passages over more than 30 months. The histologic features of heterotransplanted tumors in severe combined immunodeficiency mice were essentially the same as those of the original nonlipogenic sarcoma resembling a malignant fibrous histiocytoma. Both in vitro and in vivo, the cells exhibited immunopositive reaction for mdm2 and p53 proteins. Cytogenetically, FU-DDLS-1 displayed a hypertetraploid karyotype with giant marker chromosomes composed partly of chromosome 12 material. In addition, CGH analysis demonstrated that DNA sequence copy number changes including a gain of 12q12-q21 detected in FU-DDLS-1 were essentially the same as those in the original sarcoma. The FU-DDLS-1 cell line, which exhibits the unique conventional and molecular cytogenetic characteristics of dedifferentiated liposarcoma, should be a particularly useful model for studying the molecular pathogenesis of human dedifferentiated liposarcoma.

A pilot study of physiotherapy education using videoconferencing.

We investigated the possibility of instructing people in how to assist patients with activities of daily living via videoconferencing. A videoconference system was installed in two rooms to simulate a connection between a home and an institution. The subjects were five first-year university students. First, they saw a video-tape on several general assistance procedures to help a patient with sitting up, standing up and walking with a cane. Then they assisted a simulated patient who reproduced the effects of right hemiplegia due to a stroke. They also received instructions from a physiotherapist in another room using the videoconference system. The students experienced most difficulty in assisting the patient to stand up from sitting with legs stretched in a tatami room. Their mean score in the first session for this was 34.5 points out of 45, and this increased to 38.5 in a second session, after a question and answer period. The present study shows that the delivery of rehabilitation services is feasible in remote places.

Development of a telerehabilitation system for training physiotherapists in rural areas.

The purpose of this study was to assess the effectiveness of transmitting moving pictures via the Internet for processing by a three-dimensional motion analyser. The movements of two patients were filmed by a videocamera and transmitted from a hospital to a university department of physiotherapy. After advanced analysis using a three-dimensional motion analyser, physiotherapists prepared a treatment plan. The transmitted moving pictures were clear enough for analysis. The physiotherapists in the hospital were satisfied with the results of the motion analysis and found the guidance from an expert physiotherapist valuable. In the future, a large number of video-sequences could be transmitted during the night, stored at the university and then analysed later by an expert physiotherapist.

Examining the relationships between attention deficit/hyperactivity disorder and developmental coordination disorder symptoms, and writing performance in Japanese second grade students.

The purpose of this study was to explore the relationships between attention deficit/hyperactivity disorder and developmental coordination disorder symptoms and writing performance in Japanese second grade students from regular classrooms. The second grade students (N=873) in Japanese public elementary schools participated in this study. We examined a variety of writing tasks, such as tracing, copying, handwriting (Hiragana and Katakana), and spelling (Hiragana, Katakana, and Kanji). We employed the Japanese version of the home form ADHD-rating scale (ADHD-RS) and the Japanese version of the Developmental Coordination Disorder Questionnaire (DCDQ-J) to assess the developmental characteristics of the participating children. Seven writing performance scores were submitted to a principal component analysis with a promax rotation, which yielded three composite scores (Spelling Accuracy, Tracing and Copying Accuracy, and Handwriting Fluency). A multiple regression analysis found that inattention predicted Spelling Accuracy and Handwriting Fluency and that hyperactive-impulsive predicted Handwriting Fluency. In addition, fine motor ability predicted Tracing and Copying Accuracy. The current study offered empirical evidence suggesting that developmental characteristics such as inattention and fine motor skill are related to writing difficulties in Japanese typical developing children.

Evaluation of the Japanese version of the Developmental Coordination Disorder Questionnaire as a screening tool for clumsiness of Japanese children.

Developmental Coordination Disorder (DCD) is characterized by clumsiness and coordination difficulties. DCD interferes with academic performance and participation in physical activities and psychosocial functions, such as self-esteem, cognition, or emotion, from childhood through adolescence to adulthood. DCD is a common pediatric condition and its prevalence is estimated to be 6% worldwide. Although English questionnaires are available, there is no questionnaire to identify DCD in Japan, and therefore, no information on its prevalence is available. Recently, we developed the Japanese version of the Developmental Coordination Disorder Questionnaire (DCDQ-J). The purpose of this study was to describe the applicability of the DCDQ-J for use with a community-based population of children in Japan and to investigate the relationships between coordination and attention-deficit hyperactivity disorder (ADHD) tendencies or intelligence. The DCDQ-J was completed by 6330 parents or guardians of children and adolescents. We employed the ADHD-rating scale and determined the intelligence quotient (IQ) of the children. Two-way analysis of variance showed that the scores linearly increased as the children's grades advanced in 2 subscales, namely, control during movement and fine motor. In contrast, non-linear changes were found in the scores of the general coordination subscale. The total scores of the DCDQ-J and ADHD-RS were significantly correlated, but no relationship between DCDQ-J scores and IQ was found. The DCDQ-J is expected to be a useful screening tool to identify and assess motor coordination difficulties of children in Japan and enable cross-cultural comparisons.

Reaction time of facial affect recognition in Asperger's disorder for cartoon and real, static and moving faces.

This study used a choice reaction-time paradigm to test the perceived impairment of facial affect recognition in Asperger's disorder. Twenty teenagers with Asperger's disorder and 20 controls were compared with respect to the latency and accuracy of response to happy or disgusted facial expressions, presented in cartoon or real images and in static or moving conditions. Group analysis revealed that the Asperger group did not differ significantly from the control group in speed and accuracy for both affects and in all presentation modalities. Individual analysis, however, revealed that the proportion of participants exhibiting a happy face advantage was smaller in the Asperger group than in the control group. The results did not support the notion of impairment in facial affect recognition in terms of speed and accuracy in Asperger's disorder. Findings also revealed that the absence of happy face advantage was more prevalent in individuals with Asperger's disorder.

Establishment and characterization of a novel human desmoplastic small round cell tumor cell line, JN-DSRCT-1.

The exact nature of the desmoplastic small round cell tumor (DSRCT) remains controversial. More detailed analyses might be facilitated by the establishment of permanent DSRCT cell lines. To date, however, no human DSRCT cell line has been reported. In this study, we report the establishment of a new human cell line, JN-DSRCT-1, from the pleural effusion of a 7-year-old boy with pulmonary metastasis from a typical intra-abdominal DSRCT. JN-DSRCT-1 cells were small round or spindle shaped with oval nuclei and have been maintained continuously in vitro for over 190 passages during more than 40 months. Histologic features of the heterotransplanted tumors in severe combined immunodeficiency mouse were essentially the same as those of the original DSRCT, revealing nests or clusters of small round cells embedded in an abundant desmoplastic stroma. Both in vitro and in vivo, the cells exhibited immunopositive reactions for vimentin, desmin, cytokeratins (AE1/AE3 and CAM 5.2), epithelial membrane antigen, neuron-specific antigen, and CD57 (Leu-7). JN-DSRCT-1 cells exhibited a pathognomonic t(11;22)(p13;q12) translocation by cytogenetic analysis. In addition, RT-PCR and sequencing analysis revealed a chimeric transcriptional message of the Ewing's sarcoma gene exon 10 fused to the Wilms' tumor gene exon 8. To our knowledge, this is the first permanent human DSRCT cell line. The JN-DSRCT-1 cell line, which exhibits the unique morphologic and genetic characteristics of DSRCT, will be extremely useful for a variety of important studies such as the pathogenic mechanism, biologic behavior, and therapeutic model of human DSRCT.