Changes in physical activity level during hospitalization in patients with stroke and those with fracture: a prospective longitudinal study.

[Purpose] To examine changes in physical activity levels between admission and discharge in patients hospitalized after stroke and fracture. [Participants and Methods] Patients with stroke (n=36) or fracture (n=41) wore an accelerometer during the daytime for three days after admission and before discharge. Physical activity was divided into sedentary behavior (SB), light-intensity (LIPA), and moderate-to-vigorous (MVPA), and then compared between hospital admission and discharge using the Wilcoxon signed-rank test. The characteristics of patients with or without changes in SB during hospitalization were compared using the Mann-Whitney U test. [Results] The median LIPA time in patients after stroke and fracture increased from 107.5 and 106.7 minutes on admission to 122.0 and 127.3 minutes at discharge, and the median MVPA time increased from 2.7 and 0.7 minutes on admission to 4.2 and 2.7 minutes at discharge, respectively. In particular, LIPA in non-therapy time increased for patients both after stroke and fracture. No differences in characteristics were observed between with or without changes in SB regardless of differences in diagnoses. [Conclusion] These findings indicate that while physical activity levels increased during hospitalization, they remained below World Health Organization recommendations for MVPA, and patient characteristics alone may not account for increased activity levels.

Automatic generation of a holographically shaped beam in an actual optical system for use in material laser processing.

In-system optimization involves designing a computer-generated hologram (CGH) in an actual optical system. An important advantage of this approach is automatic generation of a target shaped beam with compensation for imperfections in the actual optical system that would degrade the reconstruction performance. We developed a novel in-system optimization method for beam shaping based on our previous research where it had been applied only to generate parallel focused beams. The key point in the application to beam shaping is to accurately express the conditions and coordinates of the actual optical system in the CGH calculation.

Soluble LR11 as a Novel Biomarker in Acute Kawasaki Disease.

BACKGROUND: Intimal smooth muscle cells (SMCs) play an important role in the vasculitis caused by Kawasaki disease (KD). Lipoprotein receptor 11 (LR11) is a member of the low-density lipoprotein receptor family, which is expressed markedly in intimal vascular SMCs and secreted in a soluble form (sLR11). sLR11 has been recently identified as a potential vascular lesion biomarker. sLR11 is reportedly elevated in patients with coronary artery lesions long after KD, but there is no description of sLR11 in acute KD. Our aim was to determine the sLR11 dynamics in acute KD and to assess its usefulness as a biomarker.Methods and Results: 106 acute KD patients and 18 age-matched afebrile controls were enrolled. KD patients were classified into the following subgroups: intravenous immunoglobulin (IVIG) responders (n=85) and non-responders (n=21). Serum sLR11 levels before IVIG therapy were higher in non-responders (median, 19.6 ng/mL; interquartile range [IQR], 13.0-24.9 ng/mL) than in controls (11.9 ng/mL, 10.4-14.9 ng/mL, P<0.01) or responders (14.3 ng/mL, 11.7-16.5 ng/mL, P<0.01). Using a cutoff of >17.5 ng/mL, non-responders to initial IVIG therapy were identified with 66.7% sensitivity and 78.8% specificity. CONCLUSIONS: sLR11 can reflect the state of acute KD and might be a biomarker for patient response to IVIG therapy.

[A Case of Pharyngolaryngitis Requiring Hospitalization during Chemotherapy for Breast Cancer].

A 45-year-old woman underwent primary systemic therapy for left breast cancer(cT1N1M0, cStage ⅡA, Luminal B [HER2-positive]). She received EC therapy(epirubicin 90mg/m2, and cyclophosphamide 900mg/m2). On the 4th and 5th day of the third EC cycle, she developed sore throat and a fever of over 38℃, and was not able to consume anything orally. She visited our hospital and underwent a laryngeal endoscopy on the 8th day of the third EC cycle, which revealed severe inflammation of her pharynx and larynx. Viral pharyngolaryngitis was suspected and hence, she was admitted to our hospital. She developed laryngeal edema after hospitalization, for which hydrocortisone was administered. She was discharged from the hospital when her symptoms improved.

Sensitivity to change and responsiveness of the Balance Evaluation Systems Test (BESTest), Mini-BESTest, and Brief-BESTest in patients with subacute cerebral infarction.

[Purpose] To compare the sensitivity to change and responsiveness of the Balance Evaluation Systems Test, Mini-Balance Evaluation Systems Test, and Brief-Balance Evaluation Systems Test in patients with subacute cerebral infarction. [Participants and Methods] Thirty patients with subacute cerebral infarction participated in this study. The Balance Evaluation Systems Test, Mini-Balance Evaluation Systems Test, Brief-Balance Evaluation Systems Test, Berg Balance Scale, and ambulatory ability were assessed on admission and discharge. Sensitivity to change was calculated using the effect size, standardized response mean, and relative efficiency. Responsiveness was analyzed by comparing the ability of the difference between the scores of the balance assessments at admission and discharge in classifying the participants' ambulatory independence. [Results] All assessments showed significant improvement from admission to discharge. The effect size of the three versions of the Balance Evaluation Systems Test ranged from 0.41 to 0.69. The standardized response mean ranged from 0.75 to 1.28. The cutoff score was 16.7% for the Balance Evaluation Systems Test, 5.5 points for the Mini-Balance Evaluation Systems Test, 1.5 points for the Brief-Balance Evaluation Systems Test, and 3.5 points for the Berg Balance Scale. [Conclusion] The sensitivity to change of the three versions of the Balance Evaluation Systems Test was high or moderate. However, the Mini-Balance Evaluation Systems Test had the highest responsiveness, as determined with the extent of ambulatory independence.

Isobutanol production in Corynebacterium glutamicum: Suppressed succinate by-production by pckA inactivation and enhanced productivity via the Entner-Doudoroff pathway.

On the basis of our previous studies of microbial L-valine production under oxygen deprivation, we developed isobutanol-producing Corynebacterium glutamicum strains. The artificial isobutanol synthesis pathway was composed of the first three steps of the L-valine synthesis pathway; and the subsequent Ehrlich Pathway: pyruvate was converted to 2-ketoisovalerate in the former reactions; and the 2-keto acid was decarboxylated into isobutyraldehyde, and subsequently reduced into isobutanol in the latter reactions. Although there exists redox cofactor imbalance in the overall reactions, i.e., NADH is generated via glycolysis whereas NADPH is required to synthesize isobutanol, it was resolved by taking advantage of the NAD-preferring mutant acetohydroxy acid isomeroreductase encoded by ilvCTM and the NAD-specific alcohol dehydrogenase encoded by adhA. Each enzyme activity to synthesize isobutanol was finely tuned by using two kinds of lac promoter derivatives. Efficient suppression of succinate by-production and improvement of isobutanol yield resulted from inactivation of pckA, which encodes phosphoenolpyruvate carboxykinase, whereas glucose consumption and isobutanol production rates decreased because of the elevated intracellular NADH/NAD+ ratio. On the other hand, introduction of the exogenous Entner-Doudoroff pathway effectively enhanced glucose consumption and productivity. Overexpression of phosphoenolpyruvate:carbohydrate phosphotransferase system specific to glucose and deletion of ilvE, which encodes branched-chain amino acid transaminase, further suppressed by-products and improved isobutanol productivity. Finally, the produced isobutanol concentration reached 280 mM at a yield of 84% (mol/mol glucose) in 24 h.

In-system optimization of a hologram for high-stability parallel laser processing.

A method for optimizing a computer-generated hologram (CGH) for high-stability laser processing is proposed. The CGH is optimized during laser processing; therefore, unpredicted dynamic changes in the laser processing system, in addition to its static imperfections, are automatically compensated for by exploiting the rewritable capability of the spatial light modulator. Consequently, the short-term and long-term stability are improved, which will contribute to the realization of high-speed, high-precision laser processing. A CGH that generated 36 parallel beams was continuously optimized, and the maximum uniformity reached 0.98, which is higher than reported in previous research. To the best of our knowledge, this is the first demonstration of gradual improvement of parallel laser processing with in-process optimization of the CGH. Furthermore, it was also demonstrated that the performance of the laser processing system against unexpected disturbances was improved.

Phototropin perceives temperature based on the lifetime of its photoactivated state.

Living organisms detect changes in temperature using thermosensory molecules. However, these molecules and/or their mechanisms for sensing temperature differ among organisms. To identify thermosensory molecules in plants, we investigated chloroplast positioning in response to temperature changes and identified a blue-light photoreceptor, phototropin, that is an essential regulator of chloroplast positioning. Based on the biochemical properties of phototropin during the cellular response to light and temperature changes, we found that phototropin perceives temperature based on the temperature-dependent lifetime of the photoactivated chromophore. Our findings indicate that phototropin perceives both blue light and temperature and uses this information to arrange the chloroplasts for optimal photosynthesis. Because the photoactivated chromophore of many photoreceptors has a temperature-dependent lifetime, a similar temperature-sensing mechanism likely exists in other organisms. Thus, photoreceptors may have the potential to function as thermoreceptors.

Electron Microscopic and Immunohistochemical Findings of the Epidermal Basement Membrane in Two Families with Nail-patella Syndrome.

Nail-patella syndrome is an autosomal dominant disorder characterized by nail dysplasia and skeletal anomaly. Some patients have been shown to have ultrastructural abnormalities of the glomerular basement membrane that result in nephrosis. However, little has been reported on the epidermal basement membrane in this condition. This paper reports 2 families with nail-patella syndrome. Direct sequencing analysis of LMX1B revealed that family 1 and family 2 were heterozygous for the mutations c.140-1G>C and c.326+1G>C, respectively. To evaluate the epidermal basement membrane zone, ultrastructural and immunohistochemical analyses were performed using skin specimens obtained from the dorsal thumb. Electron microscopy showed intact hemidesmosomes, lamina lucida, lamina densa, and anchoring fibrils. Immunofluorescence studies with antibodies against components of the epidermal basement membrane zone revealed a normal expression pattern among the components, including type IV collagen. These data suggest that nail dysplasia in patients with nail-patella syndrome is not caused by structural abnormalities of the epidermal basement membrane.

Three-dimensional stimulation and imaging-based functional optical microscopy of biological cells.

A new type of functional optical microscope system called three-dimensional (3D) stimulation and imaging-based functional optical microscopy (SIFOM) is proposed, to the best of our knowledge. SIFOM can precisely stimulate user-defined targeted biological cells and can simultaneously record the volumetric fluorescence distribution in a single acquisition. Precise and simultaneous stimulation of fluorescent-labeled biological cells is achieved by multiple 3D spots generated by digital holograms displayed on a phase-mode spatial light modulator. Single-shot 3D acquisition of the fluorescence distribution is accomplished by common-path off-axis incoherent digital holographic microscopy in which a diffraction grating with a focusing lens is displayed on another phase-mode spatial light modulator. The effectiveness of the proposed functional microscope system was verified in experiments using fluorescent microbeads and human lung cancer cells located at various defocused positions. The system can be used for manipulating the states of cells in optogenetics.

Electrical Stimulation to the Infraspinatus on Hypertrophy and Strength of the Shoulder.

The purpose of this study was to determine the effects of six weeks of electrical muscle stimulation (EMS) on the strength and muscle mass of the infraspinatus muscle. Twenty non-athletes (age: 24±3.4 years, height: 171.5±5.6 cm, mass: 65.2±8.1 kg) were randomly classified into two groups, an electrical muscle stimulation group (EMS group) and a control group (CON group). The EMS group completed a total of 18 20- min EMS sessions, three times per week over a period of six weeks, while the CON group received no intervention. The muscle thicknesses of both the infraspinatus and the deltoid muscles, the cross-sectional area (CSA) of the whole infraspinatus muscle, and the isometric and isokinetic peak torques of shoulder external rotations were measured before and after intervention. It was found that the muscle thickness of the superior infraspinatus (Pre 0.92±0.19 cm2, Post 0.99±0.16 cm2, p=0.02) and the CSA (Pre 10.99±1.32 cm2, Post 11.99±1.02 cm2, p=0.03) significantly increased in the EMS group. This study demonstrated that EMS of the infraspinatus muscle over a period of six weeks resulted in hypertrophy of the infraspinatus muscle.

Effective stretching position for the supraspinatus muscle evaluated by shear wave elastography in vivo.

BACKGROUND: Stretching is useful for increasing flexibility in clinical and athletic situations. Although several authors have recommended various stretching techniques for the supraspinatus muscle, there is no consensus on the effective stretching position owing to a lack of quantitative analysis in vivo. This study used ultrasonic shear wave elastography in vivo to verify the effective stretching positions for the supraspinatus muscle. METHODS: The study participants were 15 healthy male volunteers. The shear elastic modulus, used as the index of supraspinatus muscle elongation, was computed using ultrasonic shear wave elastography. The shear elastic modulus was measured at neutral position and maximum internal rotation in 9 positions: 0° elevation, 90° abduction, 90° flexion, maximum extension, maximum horizontal adduction at 45° and 90° elevation, and maximum horizontal abduction at 20°, 45°, and 90° elevation. RESULTS: The shear elastic moduli were significantly greater in maximum internal rotation at maximum horizontal abduction with 45° and 90° elevation and maximum internal rotation at maximum extension than those in the other positions. There were no significant differences in the shear elastic moduli among these 3 positions. CONCLUSIONS: This study demonstrated that maximum internal rotation at maximum extension, maximum internal rotation at maximum horizontal abduction with 90° elevation, and maximum internal rotation at maximum horizontal abduction with 45° elevation are effective stretching positions for the supraspinatus muscle.

Structural validity of Balance Evaluation Systems Test assessed using factor and Rasch analyses in patients with stroke.

[Purpose] The Balance Evaluation Systems Test (BESTest) is a comprehensive assessment tool, although it is not confined for use in stroke patients. This study aimed to determine the structural validity of the BESTest in self-ambulatory patients with stroke using both factor and Rasch analyses. [Participants and Methods] This retrospective study included 140 self-ambulatory patients with stroke. The structural validity of the BESTest was analyzed according to principal component, exploratory factor, Rasch, confirmatory factor, and correlation analyses. [Results] The analytical results supported a four-factor model comprising 25 items. The four factors included dynamic postural control with gait, static postural control, stepping reaction, and stability limits in sitting. Evidence of high structural validity and reliable internal consistency suggested that the 25-item BESTest is valid and reliable. Each factor was significantly correlated with lower extremity motor function and walking ability. [Conclusion] Eleven items in the BESTest were poorly correlated, and the remaining 25 items were grouped into four factors that demonstrated good structural validity for patients with stroke. Further studies should validate the applicability of the 25-item BESTest four-factor model in a larger sample of patients with stroke in a clinical setting.

Enhanced Glucose Consumption and Organic Acid Production by Engineered Corynebacterium glutamicum Based on Analysis of a pfkB1 Deletion Mutant.

In the analysis of a carbohydrate metabolite pathway, we found interesting phenotypes in a mutant strain of Corynebacterium glutamicum deficient in pfkB1, which encodes fructose-1-phosphate kinase. After being aerobically cultivated with fructose as a carbon source, this mutant consumed glucose and produced organic acid, predominantly l-lactate, at a level more than 2-fold higher than that of the wild-type grown with glucose under conditions of oxygen deprivation. This considerably higher fermentation capacity was unique for the combination of pfkB1 deletion and cultivation with fructose. In the metabolome and transcriptome analyses of this strain, marked intracellular accumulation of fructose-1-phosphate and significant upregulation of several genes related to the phosphoenolpyruvate:carbohydrate phosphotransferase system, glycolysis, and organic acid synthesis were identified. We then examined strains overexpressing several of the identified genes and demonstrated enhanced glucose consumption and organic acid production by these engineered strains, whose values were found to be comparable to those of the model pfkB1 deletion mutant grown with fructose. l-Lactate production by the ppc deletion mutant of the engineered strain was 2,390 mM (i.e., 215 g/liter) after 48 h under oxygen deprivation, which was a 2.7-fold increase over that of the wild-type strain with a deletion of ppc IMPORTANCE: Enhancement of glycolytic flux is important for improving microbiological production of chemicals, but overexpression of glycolytic enzymes has often resulted in little positive effect. That is presumably because the central carbon metabolism is under the complex and strict regulation not only transcriptionally but also posttranscriptionally, for example, by the ATP/ADP ratio. In contrast, we studied a mutant strain of Corynebacterium glutamicum that showed markedly enhanced glucose consumption and organic acid production and, based on the findings, identified several genes whose overexpression was effective in enhancing glycolytic flux under conditions of oxygen deprivation. These results will further understanding of the regulatory mechanisms of glycolytic flux and can be widely applied to the improvement of the microbial production of useful chemicals.

Role of HER2-Related Biomarkers (HER2, p95HER2, HER3, PTEN, and PIK3CA) in the Efficacy of Lapatinib plus Capecitabine in HER2-Positive Advanced Breast Cancer Refractory to Trastuzumab.

OBJECTIVE: The aim of this study was to investigate the correlation between human epidermal growth factor receptor 2 (HER2)-related biomarkers and the treatment outcomes using lapatinib plus capecitabine (LC) and to evaluate the influence of the estrogen receptor (ER) status in trastuzumab-refractory HER2-positive advanced breast cancer. METHOD: Eighty patients were enrolled in this study. Total HER2, p95HER2, and total HER3 expression were quantified using the VeraTag assays. PTEN (phosphatase and tensin homolog) and p95 expression was evaluated using immunohistochemistry and PIK3CA mutation using direct sequencing. RESULTS: The response rate to LC was 30%, clinical benefit rate was 51.3%, and the median progression-free survival (PFS) was 174.5 days. ER negativity significantly correlated with higher HER2 and p95HER2. The lower HER2 and PIK3CA mutations were often observed in the nonresponders. A high p95HER2 expression correlated with longer PFS especially in the high HER2- and ER-positive cases. Patients without the PIK3CA mutation showed longer PFS in the same subset. Overall survival after LC significantly correlated with the number of recurrence organs. CONCLUSION: LC therapy is effective in trastuzumab-refractory HER2-positive breast cancer. Moreover, the biomarker expression differed depending on ER status, and a high p95HER2 expression and wild-type PIK3CA gene correlated with longer PFS especially in the ER-positive cases.

Effect of Capacitive and Resistive electric transfer on haemoglobin saturation and tissue temperature.

PURPOSE: This study aims to evaluate the effects of Capacitive and Resistive electric transfer (CRet) and hotpack (HP) on haemoglobin saturation and tissue temperature. MATERIALS AND METHODS: The participants were 13 healthy males (mean age 24.5 ± 3.0). They underwent three interventions on different days: (1) CRet (CRet group), (2) HP (HP group) and (3) CRet without power (sham group). The intervention and measurement were applied at the lower paraspinal muscle. Indiba® active ProRecovery HCR902 was used in the CRet group, and the moist heat method was used in the HP group. Oxygenated, deoxygenated and total haemoglobin (oxy-Hb, deoxy-Hb, total-Hb) counts were measured before and after the 15-min interventions, together with the temperature at the skin surface, and at depths of 10 mm and 20 mm (ST, 10mmDT and 20mmDT, respectively). The haemoglobin saturation and tissue temperature were measured until 30 min after the intervention and were collected at 5-min intervals. Statistical analysis was performed for each index by using the Mann-Whitney U test for comparisons between all groups at each time point. RESULTS: Total-Hb and oxy-Hb were significantly higher in the CRet group than in the HP group continuously for 30 min after the intervention. The 10mmDT and 20mmDT were significantly higher in the CRet group than in the HP group from 10- to 30 min after intervention. CONCLUSIONS: The effect on haemoglobin saturation was higher in the CRet group than in the HP group. In addition, the CRet intervention warmed deep tissue more effectively than HP intervention.

Functional analysis of arabinofuranosidases and a xylanase of Corynebacterium alkanolyticum for arabinoxylan utilization in Corynebacterium glutamicum.

Xylooligosaccharides (XOSs) and arabinoxylooligosaccharides (AXOSs) are major oligosaccharides derived from arabinoxylan. In our previous report, Corynebacterium glutamicum was engineered to utilize XOSs by introducing Corynebacterium alkanolyticum xyloside transporter and ß-xylosidase. However, this strain was unable to consume AXOSs due to the absence of α-L-arabinofuranosidase activity. In this study, to confer AXOS utilization ability on C. glutamicum, two putative arabinofuranosidase genes (abf51A and abf51B) were isolated from C. alkanolyticum by the combination of degenerate PCR and genome walking methods. Recombinant Abf51A and Abf51B heterologously expressed in Escherichia coli showed arabinofuranosidase activities toward 4-nitrophenyl-α-L-arabinofuranoside with k cat values of 150 and 63, respectively, with optimum at pH 6.0 to 6.5. However, Abf51A showed only a slight activity toward AXOSs and was more susceptible to product inhibition by arabinose and xylose than Abf51B. Introduction of abf51B gene into the C. glutamicum XOS-utilizing strain enabled it to utilize AXOSs as well as XOSs. The xylI gene encoding a putative xylanase was found upstream of the C. alkanolyticum xyloside transporter genes. A signal peptide was predicted at the N-terminus of the xylI-encoding polypeptide, which indicated XylI was a secreted protein. Recombinant mature XylI protein heterologously expressed in E. coli showed a xylanase activity toward xylans from various plant sources with optimum at pH 6.5, and C. glutamicum recombinant strain expressing native XylI released xylose, xylobiose, xylotriose, and arabino-xylobiose from arabinoxylan. Finally, introduction of the xylI gene into the C. glutamicum AXOS-utilizing strain enabled it to directly utilize arabinoxylan.

Temperature-dependent signal transmission in chloroplast accumulation response.

Chloroplast photorelocation movement, well-characterized light-induced response found in various plant species from alga to higher plants, is an important phenomenon for plants to increase photosynthesis efficiency and avoid photodamage. The signal for chloroplast accumulation movement connecting the blue light receptor, phototropin, and chloroplasts remains to be identified, although the photoreceptors and the mechanism of movement via chloroplast actin filaments have now been revealed in land plants. The characteristics of the signal have been found; the speed of signal transfer is about 1 µm min-1 and that the signal for the accumulation response has a longer life and is transferred a longer distance than that of the avoidance response. Here, to collect the clues of the unknown signal substances, we studied the effect of temperature on the speed of signal transmission using the fern Adiantum capillus-veneris and found the possibility that the mechanism of signal transfer was not dependent on the simple diffusion of a substance; thus, some chemical reaction must also be involved. We also found new insights of signaling substances, such that microtubules are not involved in the signal transmission, and that the signal could even be transmitted through the narrow space between chloroplasts and the plasma membrane.

Isometric muscle activation of the serratus anterior and trapezius muscles varies by arm position: a pilot study with healthy volunteers with implications for rehabilitation.

BACKGROUND: This study aimed to determine the most appropriate angle and moving direction of the arm for improving coordination of the periscapular muscles, including the serratus anterior (SA), upper trapezius (UT), middle trapezius (MT), and lower trapezius (LT). METHODS: Muscle activation amplitudes were evaluated in the SA, UT, MT, and LT in 11 healthy subjects by use of surface electromyography. The subjects were asked to maintain the arm position at 5 elevated positions with maximal effort against applied manual forces, which were directed from upper to lower (test 1), lower to upper (test 2), posterior to anterior in the frontal plane and lateral to medial in the sagittal plane (test 3), and anterior to posterior in the frontal plane and medial to lateral in the sagittal plane (test 4). The relative activity of the UT with respect to the SA, MT, and LT was calculated, resulting in the UT/SA, UT/MT, and UT/LT ratios. RESULTS: Test 4 in all positions but 150° of elevation in the frontal plane showed high activity of the SA with a low UT/SA ratio. High MT activity with a low UT/MT ratio was observed during test 3 at the 90° elevated position, whereas high LT activity without UT hyperactivation was not found. DISCUSSION: To strengthen the periscapular muscles in the balanced condition, horizontal adduction is recommended for the SA. Horizontal abduction at the 90° elevated position should be effective for the MT. Because no technique in this study was effective for the LT, further studies are needed.

Gain-of-function mutations in RIT1 cause Noonan syndrome, a RAS/MAPK pathway syndrome.

RAS GTPases mediate a wide variety of cellular functions, including cell proliferation, survival, and differentiation. Recent studies have revealed that germline mutations and mosaicism for classical RAS mutations, including those in HRAS, KRAS, and NRAS, cause a wide spectrum of genetic disorders. These include Noonan syndrome and related disorders (RAS/mitogen-activated protein kinase [RAS/MAPK] pathway syndromes, or RASopathies), nevus sebaceous, and Schimmelpenning syndrome. In the present study, we identified a total of nine missense, nonsynonymous mutations in RIT1, encoding a member of the RAS subfamily, in 17 of 180 individuals (9%) with Noonan syndrome or a related condition but with no detectable mutations in known Noonan-related genes. Clinical manifestations in the RIT1-mutation-positive individuals are consistent with those of Noonan syndrome, which is characterized by distinctive facial features, short stature, and congenital heart defects. Seventy percent of mutation-positive individuals presented with hypertrophic cardiomyopathy; this frequency is high relative to the overall 20% incidence in individuals with Noonan syndrome. Luciferase assays in NIH 3T3 cells showed that five RIT1 alterations identified in children with Noonan syndrome enhanced ELK1 transactivation. The introduction of mRNAs of mutant RIT1 into 1-cell-stage zebrafish embryos was found to result in a significant increase of embryos with craniofacial abnormalities, incomplete looping, a hypoplastic chamber in the heart, and an elongated yolk sac. These results demonstrate that gain-of-function mutations in RIT1 cause Noonan syndrome and show a similar biological effect to mutations in other RASopathy-related genes.