Identifying preventative measures against frailty, locomotive syndrome, and sarcopenia in young adults: a pilot study.

[Purpose] The purpose of this study was to propose potential preventive measures against future mobility impairments and muscle loss in healthy young adults by comprehensively evaluating their status of frailty, locomotive syndrome, and sarcopenia. [Participants and Methods] A total of 83 Japanese young adults were enrolled in this study and evaluated using the diagnostic criteria for frailty (Japanese version of the Cardiovascular Health Study), locomotive syndrome (calculated as scores of the LOCOMO-25, the stand-up test, and two-step test), and sarcopenia (handgrip strength, usual gait test, skeletal muscle index). [Results] The prevalence of frailty-prefrailty (45.9%) was higher than that of presarcopenia (22.3%) and locomotive syndrome (14.1%). The prevalence of combinations of frailty-prefrailty and presarcopenia; frailty-prefrailty and locomotive syndrome; and frailty-prefrailty, locomotive syndrome, and pre-sarcopenia was 9.4%, 4.7%, and 3.5%, respectively. [Conclusion] Only 40% of the participants did not meet the three diagnostic criteria. On the other hand, there were not many participants with sarcopenia and/or locomotive syndrome diagnoses, suggesting that the improvement in intrinsic skeletal muscle mass rather than physical function is important for healthy Japanese young adults and could possibly be protective against future mobility or muscle-loss disorders.

Is blood flow-restricted training effective for rehabilitation of a pianist with residual neurological symptoms in the upper limbs? A case study.

[Purpose] We investigated whether blood flow-restricted training known as KAATSU training, was effective for rehabilitation of a pianist with residual neurological symptoms in the upper limbs. [Participant and Methods] A pianist with residual neurological symptoms in the upper body played "Revolutionary Etude" under two conditions: piano performance with (Piano-blood flow-restricted) and without (Piano-control) the restriction of blood flow to the upper limbs. In the Piano-blood flow-restricted exercise, a pressure of 130-170 mmHg was applied around the most proximal portion of both arms. The changes in upper limb circumference and muscle strength were measured before, immediately after, and 15 min after the performance. The impression of the piano performance was recorded after the Piano-blood flow-restricted exercise. [Results] Immediately after the piano performance, the forearm and upper arm circumferences had increased significantly in both arms, and the change was greater in the Piano-blood flow-restricted than in the Piano-control condition. The handgrip strength for the right arm also showed greater changes in the former than the latter. However, there were no significant differences between the two conditions regarding the handgrip strength of the left arm. [Conclusion] There is a high possibility that blood flow-restricted training is effective for rehabilitation of the pianist with residual neurological symptoms in the upper limbs.

Application of Ionic Liquids to Energy Storage and Conversion Materials and Devices.

Ionic liquids (ILs) are liquids consisting entirely of ions and can be further defined as molten salts having melting points lower than 100 °C. One of the most important research areas for IL utilization is undoubtedly their energy application, especially for energy storage and conversion materials and devices, because there is a continuously increasing demand for clean and sustainable energy. In this article, various application of ILs are reviewed by focusing on their use as electrolyte materials for Li/Na ion batteries, Li-sulfur batteries, Li-oxygen batteries, and nonhumidified fuel cells and as carbon precursors for electrode catalysts of fuel cells and electrode materials for batteries and supercapacitors. Due to their characteristic properties such as nonvolatility, high thermal stability, and high ionic conductivity, ILs appear to meet the rigorous demands/criteria of these various applications. However, for further development, specific applications for which these characteristic properties become unique (i.e., not easily achieved by other materials) must be explored. Thus, through strong demands for research and consideration of ILs unique properties, we will be able to identify indispensable applications for ILs.

Field-Based Simplified Approach of Evaluating Knee Extensor Muscle Strength and Size in University Freshmen Women.

Context: Periodic assessment of knee extensor muscle strength and size is important for all ages to evaluate the functional status of individuals and to identify and treat those at risk for mobility problems and frailty; however, it is not fully understood whether these field-based simplified approaches correspond to evaluation in knee extensor muscle strength or size. Objective: To examine the relationship between field-based simplified evaluation approaches and knee extensor muscle strength or size in young women. Design: Experimental. Setting: University research laboratory. Subjects: A total of 62 university freshmen women volunteered to participate in this study. Main Outcome Measures: Knee extensor muscle thickness was measured at the anterior half of thigh length; muscle strength was measured when subjects performed knee extension. Field-based simplified approaches (sit-to-stand, standing long jump, handgrip, and upper leg 50% [thigh] girth) were also measured. Results: Maximal strength was correlated with thigh girth, handgrip, and standing long jump, but not with the sit-to-stand test. Muscle thickness was correlated with thigh girth and handgrip, but not with standing long jump or the sit-to-stand test. A stepwise multiple-regression analysis was calculated using the predictor thigh girth and standing long jump to predict knee extensor maximal strength (R2 = .295). To predict knee extensor muscle thickness, the predictor thigh girth was calculated (R2 = .202). Conclusions: Knee extensor muscle strength and size could be evaluated by the field-based simplified approaches, in particular by the thigh girth measurement, which may be a major determinant to maintain activities of daily living for healthy young women. However, the 4 field-based simplified approaches appear to be still not of high impact.

Key factor governing the physicochemical properties and extent of proton transfer in protic ionic liquids: ΔpKa or chemical structure?

In order to identify the key factor governing the transport properties and extent of proton transfer in protic ionic liquids (PILs), a series of PILs were prepared by simple neutralisation of a super-strong acid, bis(trifluoromethanesulfonyl)amide acid (H[NTf2]), with a range of amines comprising diverse structures, including secondary and tertiary amines. The bulk physicochemical properties of the resulting PILs with a wide variation in the ΔpKa values of the constituent acid and protonated bases were compared to those of previously reported PILs derived from a super-strong base, 1,8-diazabicyclo[5.4.0]-7-undecene (DBU), and different acids (M. S. Miran, H. Kinoshita, T. Yasuda, M. A. B. H. Susan and M. Watanabe, Phys. Chem. Chem. Phys., 2012, 14, 5178-5186) with emphasis on the thermal stability, vibrational spectroscopy, density, viscosity, conductivity, and ionicity characteristics. The thermal stability of all PILs, as analysed by thermogravimetric analysis (TGA), was found to linearly increase with the ΔpKa values. However, isothermal TGA measurements for an extended duration indicated that [NTf2]-based PILs exhibit higher thermal stability than [DBU]-based PILs at low ΔpKa values (<15). PILs with secondary amines showed higher viscosity and lower ionic conductivity than those with tertiary amines because of the formation of multiple hydrogen bonds. The evaluated ionicity suggested that all [NTf2]-based PILs are "good PILs", irrespective of the basicity of the amines, and have higher ionicity than [DBU]-based PILs particularly at low ΔpKa values (<15).

Amphoteric water as acid and base for protic ionic liquids and their electrochemical activity when used as fuel cell electrolytes.

Amphoteric water was mixed with equimolar amounts of a super-strong acid, trifluoromethanesulfonic acid (TfOH), and a super-strong base, 1,8-diazabicyclo[5.4.0]-7-undecene (DBU). Bulk physicochemical and electrochemical properties of the mixtures were compared with those of the best ever reported protic ionic liquid (PIL), diethylmethylammonium trifluoromethanesulfonate ([dema][TfO]), which has excellent physicochemical properties as a fuel cell electrolyte. The acidic mixture ([H3O][TfO]) behaved as a protic ionic liquid, while the basic mixture ([DBU]OH) showed incomplete proton transfer. The Walden plot indicated that [H3O][TfO] behaves as a good PIL, similar to [dema][TfO], whereas [DBU]OH behaves as a poor PIL. [H3O][TfO] showed excellent H2/O2 fuel cell performance at 80 °C; however, the performance deteriorated as the bulk water content increased, because of the retardation of the electrode kinetics due to the oxidation of Pt in the presence of bulk water. On the other hand, [DBU]OH exhibited very poor performance possibly because of the existence of neutral species in the system.

Short physical performance battery for middle-aged and older adult cardiovascular disease patients: implication for strength tests and lower extremity morphological evaluation.

[Purpose] To examine if the SPPB is higher with healthy subjects than outpatients, which was higher than inpatients and if the SPPB can be validated assessment tool for strength tests and lower extremity morphological evaluation in cardiovascular disease patients. [Subjects and Methods] Twenty-four middle aged and older adults with cardiovascular disease were recruited from inpatient and outpatient facilities and assigned to separate experimental groups. Twelve age-matched healthy volunteers were assigned to a control group. SPPB test was used to assess balance and functional motilities. The test outcomes were compared with level of care (inpatient vs. outpatient), physical characteristics, strength and lower extremity morphology. [Results] Total SPPB scores, strength tests (knee extensor muscle strength), and lower extremity morphological evaluation (muscle thickness of anterior and posterior mid-thigh and posterior lower-leg) were greater in healthy subjects and outpatients groups compared with inpatients. To predict total Short Physical Performance Battery scores, the predicted knee extension and anterior mid-thigh muscle thickness were calculated. [Conclusion] The SPPB is an effective tool as the strength tests and lower extremity morphological evaluation for middle-aged and older adult cardiovascular disease patients. Notably, high knee extensor muscle strength and quadriceps femoris muscle thickness are positively associated with high SPPB scores.

Repetitive restriction of muscle blood flow enhances mTOR signaling pathways in a rat model.

Skeletal muscle is a plastic organ that adapts its mass to various stresses by affecting pathways that regulate protein synthesis and degradation. This study investigated the effects of repetitive restriction of muscle blood flow (RRMBF) on microvascular oxygen pressure (PmvO2), mammalian target of rapamycin (mTOR) signaling pathways, and transcripts associated with proteolysis in rat skeletal muscle. Eleven-week-old male Wistar rats under anesthesia underwent six RRMBF consisting of an external compressive force of 100 mmHg for 5 min applied to the proximal portion of the right thigh, each followed by 3 min rest. During RRMBF, PmvO2 was measured by phosphorescence quenching techniques. The total RNA and protein of the tibialis anterior muscle were obtained from control rats, and rats treated with RRMBF 0-6 h after the stimuli. The protein expression and phosphorylation of various signaling proteins were determined by western blotting. The mRNA expression level was measured by real-time RT-PCR analysis. The total muscle weight increased in rats 0 h after RRMBF, but not in rats 1-6 h. During RRMBF, PmvO2 significantly decreased (36.1 ± 5.7 to 5.9 ± 1.7 torr), and recovered at rest period. RRMBF significantly increased phosphorylation of p70 S6-kinase (p70S6k), a downstream target of mTOR, and ribosomal protein S6 1 h after the stimuli. The protein level of REDD1 and phosphorylation of AMPK and MAPKs did not change. The mRNA expression levels of FOXO3a, MuRF-1, and myostatin were not significantly altered. These results suggested that RRMBF significantly decreased PmvO2, and enhanced mTOR signaling pathways in skeletal muscle using a rat model, which may play a role in diminishing muscle atrophy under various conditions in human studies.

Inferring the global structure of chromosomes from structural variations.

BACKGROUND: Next generation sequencing (NGS) technologies have made it possible to exhaustively detect structural variations (SVs) in genomes. Although various methods for detecting SVs have been developed, the global structure of chromosomes, i.e., how segments in a reference genome are extracted and ordered in an unknown target genome, cannot be inferred by detecting only individual SVs. RESULTS: Here, we formulate the problem of inferring the global structure of chromosomes from SVs as an optimization problem on a bidirected graph. This problem takes into account the aberrant adjacencies of genomic regions, the copy numbers, and the number and length of chromosomes. Although the problem is NP-complete, we propose its polynomial-time solvable variation by restricting instances of the problem using a biologically meaningful condition, which we call the weakly connected constraint. We also explain how to obtain experimental data that satisfies the weakly connected constraint. CONCLUSION: Our results establish a theoretical foundation for the development of practical computational tools that could be used to infer the global structure of chromosomes based on SVs. The computational complexity of the inference can be reduced by detecting the segments of the reference genome at the ends of the chromosomes of the target genome and also the segments that are known to exist in the target genome.

Effect of low-load resistance exercise with and without blood flow restriction to volitional fatigue on muscle swelling.

PURPOSE: The effects on muscle swelling were compared between low-load resistance exercise to exhaustion with (BFR) and without blood flow restriction (NBFR). METHODS: Ten young men [aged 27 (SD 5) years, standing height 1.74 (SD 0.05) m, body mass 70.3 (SD 4.3) kg] performed 20 % of one repetition maximal dumbbell curl exercise to exhaustion (four sets, rest intervals were 30 s for BFR and/or 3 min for NBFR, respectively). One arm was randomly chosen for BFR exercise and the other arm performed NBFR exercise. During the BFR exercise session, an elastic cuff was worn proximally on the testing arm at 160 mmHg. Electromyography (EMG) signals were recorded from surface electrodes placed on the biceps brachii muscle and analyzed for integrated EMG (iEMG). Biceps brachii muscle thickness (MTH) was measured using B-mode ultrasound. RESULTS: The total number of exercise repetitions was greater (p < 0.01) in NBFR (221 ± 67 reps) than in BFR (111 ± 36 reps). During the exercise session, iEMG for biceps brachii muscles increased (p < 0.01) during BFR and NBFR (3.94 and 4.45 times of baseline value). Immediately after the exercise, MTH sharply increased (p < 0.01) with BFR and NBFR (1.21 and 1.20 times of baseline value). These results demonstrate that both BFR and NBFR exercises lead to pronounced muscle activation and muscle swelling. CONCLUSION: Low-load resistance exercise to exhaustion is an effective method for promoting muscle swelling regardless of BFR. Furthermore, our data indicate that the increase in muscle swelling for both NBFR and BFR is maintained even 60 min after the exercise.

Solubility of poly(methyl methacrylate) in ionic liquids in relation to solvent parameters.

The solubility of poly(methyl methacrylate) (PMMA) in 1-alkyl-3-methylimidazolium ionic liquids (ILs) with different anionic structures has been explored. Nearly monodisperse PMMA-grafted silica nanoparticles (PMMA-g-NPs) were used as a measurement probe for evaluating the PMMA solubility in ILs. The hydrodynamic radius (Rh) of PMMA-g-NPs was measured in the ILs by dynamic light scattering (DLS). Changes in Rh and colloidal stability, that is, the PMMA-solubility change in the ILs, were observed depending on the ionic structures of the ILs. The solubility was mainly affected by the anionic structures of the ILs rather than by the alkyl chain length of the cationic structure. Solvent parameters, including Lewis basicity, solubility parameters, and a hydrophobicity parameter, were used to discuss the change in the PMMA solubility in ILs with different ionic structures. By considering the PMMA solubility in the ILs using these parameters, it was found that there is a good correlation between the PMMA solubility and the hydrophobicity parameter of the anions. Although the change in the PMMA solubility with different cationic structures was not remarkable, the hydrophobicity of the cations also played a role in the solvation of PMMA by providing a low-polarity environment adequate to dissolve PMMA.

Local amplification of storm surge by Super Typhoon Haiyan in Leyte Gulf.

Typhoon Haiyan, which struck the Philippines in November 2013, was an extremely intense tropical cyclone that had a catastrophic impact. The minimum central pressure of Typhoon Haiyan was 895 hPa, making it the strongest typhoon to make landfall on a major island in the western North Pacific Ocean. The characteristics of Typhoon Haiyan and its related storm surge are estimated by numerical experiments using numerical weather prediction models and a storm surge model. Based on the analysis of best hindcast results, the storm surge level was 5-6 m and local amplification of water surface elevation due to seiche was found to be significant inside Leyte Gulf. The numerical experiments show the coherent structure of the storm surge profile due to the specific bathymetry of Leyte Gulf and the Philippines Trench as a major contributor to the disaster in Tacloban. The numerical results also indicated the sensitivity of storm surge forecast.

Effects of detraining after blood flow-restricted low-intensity training on muscle size and strength in older adults.

BACKGROUND: Although blood flow-restricted low-intensity resistance training (BFR-RT) increases muscle size and strength in older adults, the effect of detraining on muscle adaptation is unclear. METHODS: We investigated the effects of 24 weeks of detraining on thigh muscle cross-sectional area (CSA) and one-repetition maximum strength (1-RM) in older adults who had previously participated in 12 weeks of training (BFR-RT, 20-30 % 1-RM, knee extension and leg press) or non-BFR training. RESULTS: Both 1-RM and relative dynamic strengths (1-RM divided by quadriceps femoris (QF) muscle CSA) were higher at both post-training and detraining than at pre-training for the BFR-RT group (p < 0.05). QF muscle CSA was higher at only post-training than at pre-training for the BFR-RT group (p < 0.01). CONCLUSION: Increased muscle strength following 12 weeks of training with BFR-RT was well preserved at 24 weeks of detraining, which is due mainly to neural adaptation in older adults.

Effects of Acute Piano Performance With Blood Flow Restriction on Upper Limb Muscle and Perceptual Response in Pianists.

BACKGROUND:  Long-term and prolonged piano performance does not provide essential skeletal muscle training benefits while increasing the risk of injury to the upper extremities. Unlike high-intensity exercise training, moderate blood flow restriction (BFR) training has been found to improve neuromuscular mechanisms with a variety of physical exercises (machine, elastic band, walking, electrical stimulation, and body weight). AIM AND METHODS:  We investigated the physiological and perceptual responses related to piano performance with or without BFR based on acute responses of neuromuscular mechanisms. Student or professional pianists (n=7) performed the "Revolutionary Etude" on the piano with (Piano-BFR) and without (Piano-Ctrl) BFR. During the Piano-BFR performance, 150-180 mmHg of cuff pressure was applied around the most proximal region of both arms as a moderate BFR. RESULTS:  Changes in upper limb girth, muscle thickness, and hand grip strength were measured before and immediately after the performance. After the performance, perceptual and other responses were recorded. Immediately after the performance, the Piano-BFR condition induced greater changes in girth (forearm and upper arm), muscle thickness (forearm), and handgrip strength than the Piano-Ctrl condition. Piano-BFR was (p<0.01) higher than Piano-Ctrl on eight questions regarding perceptual response (upper arm fatigue and difficulty playing the piano). Piano performance with BFR was revealed to increase upper extremity muscle size and fatigue in pianists after playing. CONCLUSION:  Piano performance with BFR was revealed to increase upper extremity muscle size and fatigue in pianists after playing. The effect of BFR on neuromuscular mechanisms on piano performance was greater in the forearm than in the upper arm.

CAN WE DIAGNOSE SARCOPENIA USING ANTERIOR FEMORAL MUSCLE THICKNESS IN PATIENTS WITH CARDIOVASCULAR DISEASE?

Objective: Making the diagnosis of sarcopenia is not always easy and this is especially true for those with cardiovascular disease. The purpose of this study is to investigate whether it is possible to diagnose sarcopenia by using ultrasound-guided measurements of anterior femoral muscle thickness. Methods: We investigated the utility of ultrasound-guided measurements of anterior femoral muscle thickness in 1075 hospitalized patients with cardiovascular disease (675 men). As a comparison, sarcopenia was assessed by skeletal muscle mass index using bioelectrical impedance analysis and the Asia Working Group for Sarcopenia criteria. Results: When the receiver operating characteristic curve using muscle thickness was examined, we found this could be used to make the diagnosis of sarcopenia (men: cutoff value 2.425 cm, area under the curve 0.796; women: cutoff value 1.995 cm, area under the curve 0.746). The prevalence of sarcopenia according to the criteria with skeletal muscle mass index was 34.2% in men and 51.8% in women, while its prevalence according to the cutoff value of muscle thickness was 29.2% in men and 36.7% in women. Conclusion: Ultrasound-guided measurement of the anterior femoral muscle thickness is a simple and useful method to help make the diagnosis of sarcopenia in patients with cardiovascular disease.

Interactions in ion pairs of protic ionic liquids: comparison with aprotic ionic liquids.

The stabilization energies for the formation (E(form)) of 11 ion pairs of protic and aprotic ionic liquids were studied by MP2/6-311G** level ab initio calculations to elucidate the difference between the interactions of ions in protic ionic liquids and those in aprotic ionic liquids. The interactions in the ion pairs of protic ionic liquids (diethylmethylammonium [dema] and dimethylpropylammonium [dmpa] based ionic liquids) are stronger than those of aprotic ionic liquids (ethyltrimethylammonium [etma] based ionic liquids). The E(form) for the [dema][CF3SO3] and [dmpa][CF3SO3] complexes (-95.6 and -96.4 kcal/mol, respectively) are significantly larger (more negative) than that for the [etma][CF3SO3] complex (-81.0 kcal/mol). The same trend was observed for the calculations of ion pairs of the three cations with the Cl(-), BF4(-), TFSA(-) anions. The anion has contact with the N-H bond of the dema(+) or dmpa(+) cations in the most stable geometries of the dema(+) and dmpa(+) complexes. The optimized geometries, in which the anions locate on the counter side of the cations, are 11.0-18.0 kcal/mol less stable, which shows that the interactions in the ions pairs of protic ionic liquids have strong directionality. The E(form) for the less stable geometries for the dema(+) and dmpa(+) complexes are close to those for the most stable etma(+) complexes. The electrostatic interaction, which is the major source of the attraction in the ion pairs, is responsible for the directionality of the interactions and determining the magnitude of the interaction energy. Molecular dynamic simulations of the [dema][TFSA] and [dmpa][TFSA] ionic liquids show that the N-H bonds of the cations have contact with the negatively charged (oxygen and nitrogen) atoms of TFSA(-) anion, while the strong directionality of the interactions was not suggested from the simulation of the [etma][CF3SO3] ionic liquid.

Comparison of muscle hypertrophy following 6-month of continuous and periodic strength training.

To compare the effects of a periodic resistance training (PTR) program with those of a continuous resistance training (CTR) program on muscle size and function, 14 young men were randomly divided into a CTR group and a PTR group. Both groups performed high-intensity bench press exercise training [75 % of one repetition maximum (1-RM); 3 sets of 10 reps] for 3 days per week. The CTR group trained continuously over a 24-week period, whereas the PTR group performed three cycles of 6-week training (or retraining), with 3-week detraining periods between training cycles. After an initial 6 weeks of training, increases in cross-sectional area (CSA) of the triceps brachii and pectoralis major muscles and maximum isometric voluntary contraction of the elbow extensors and 1-RM were similar between the two groups. In the CTR group, muscle CSA and strength gradually increased during the initial 6 weeks of training. However, the rate of increase in muscle CSA and 1-RM decreased gradually after that. In the PTR group, increase in muscle CSA and strength during the first 3-week detraining/6-week retraining cycle were similar to that in the CTR group during the corresponding period. However, increase in muscle CSA and strength during the second 3-week detraining/6-week retraining cycle were significantly higher in the PTR group than in the CTR group. Thus, overall improvements in muscle CSA and strength were similar between the groups. The results indicate that 3-week detraining/6-week retraining cycles result in muscle hypertrophy similar to that occurring with continuous resistance training after 24 weeks.

Effects of high-intensity and blood flow-restricted low-intensity resistance training on carotid arterial compliance: role of blood pressure during training sessions.

We examined the effects of high-intensity resistance training (HIT) and low-intensity blood flow-restricted (LI-BFR) resistance training on carotid arterial compliance. Nineteen young men were randomly divided into HIT (n = 9) or LI-BFR (n = 10) groups. The HIT and LI-BFR groups performed 75 and 30 %, respectively, of one-repetition maximum (1-RM) bench press exercise, 3 days per week for 6 weeks. During the training sessions, the LI-BFR group wore elastic cuffs around the most proximal region of both arms. Muscle cross-sectional area (CSA), 1-RM strength, and carotid arterial compliance were measured before and 3 days after the final training session. Acute changes in systolic arterial pressure (SAP), plasma endothelin-1 (ET-1), nitrite/nitrate (NOx), and noradrenalin concentrations were also measured during and after a bout of training session. The training led to significant increases (P < 0.01) in bench press 1-RM and arm and chest muscle CSA in the two training groups. Carotid arterial compliance decreased significantly (P < 0.05) in the HIT group, but not in the LI-BFR group. There was a significant correlation (r = -0.533, P < 0.05) between the change in carotid arterial compliance and the acute change in SAP during training sessions; however, ET-1 and NOx did not correlate with carotid arterial compliance. Our results suggest that muscle CSA and strength increased following 6 weeks of both HIT and LI-BFR training. However, carotid arterial compliance decreased in only the HIT group, and the changes were correlated with SAP elevations during exercise sessions.

Complete sequencing and characterization of 21,243 full-length human cDNAs.

As a base for human transcriptome and functional genomics, we created the "full-length long Japan" (FLJ) collection of sequenced human cDNAs. We determined the entire sequence of 21,243 selected clones and found that 14,490 cDNAs (10,897 clusters) were unique to the FLJ collection. About half of them (5,416) seemed to be protein-coding. Of those, 1,999 clusters had not been predicted by computational methods. The distribution of GC content of nonpredicted cDNAs had a peak at approximately 58% compared with a peak at approximately 42%for predicted cDNAs. Thus, there seems to be a slight bias against GC-rich transcripts in current gene prediction procedures. The rest of the cDNAs unique to the FLJ collection (5,481) contained no obvious open reading frames (ORFs) and thus are candidate noncoding RNAs. About one-fourth of them (1,378) showed a clear pattern of splicing. The distribution of GC content of noncoding cDNAs was narrow and had a peak at approximately 42%, relatively low compared with that of protein-coding cDNAs.