Simulated validation of intron-less transgene detection using DELLY for gene-doping control in horse sports.

Gene doping is prohibited in horseracing. In a previous study, we developed a method for non-targeted transgene detection using DELLY, which is based on split-read (SR) and paired-end (PE) algorithms to detect structural variants, on WGS data. In this study, we validated the detection sensitivity of DELLY using artificially generated sequence data of 12 target genes. With DELLY, at least one intron was detected as a deletion in eight targeted genes using the 150 bp PE read WGS data, whereas all targeted genes were detected by DELLY using the 100 bp PE read data. The detection sensitivity was higher in 100 bp PE reads than in 150 bp PE reads, despite a lower total sequence coverage, probably because of mismatch tolerance between the mapped reads and reference genome. In addition, it was observed that the average intron size detected by SR alone was 293 bp and that that detected by both SR and PE was 8924 bp. Thus, we showed that transgenes with various intron-exon structures could be detected using DELLY, suggesting its application in gene-doping control in horses.

Tracheostomy can fatally exacerbate sleep-disordered breathing in multiple system atrophy.

Tracheostomy has been employed to release the airway obstruction at the glottic level and to prevent sudden death in patients with multiple system atrophy (MSA). However, sudden death is possible even after tracheostomy. Nocturnal polysomnography showed that the apnea-hypopnea index became higher after tracheostomy, and all tracheostomized patients had frequent central sleep apneas.

CSF tau protein: a new prognostic marker for Guillain-Barré syndrome.

We measured the CSF tau protein levels in 26 patients with Guillain-Barré syndrome. The levels of the poor outcome group (Hughes grade at 6 months was between II and VI, n = 6) were higher than those of the good outcome group (0 or I, n = 20) (p < 0.0005). The higher levels of CSF tau may reflect axonal degeneration and could predict a poor clinical outcome in Guillain-Barré syndrome.

The female-killing chromosome of the silkworm, Bombyx mori, was generated by translocation between the Z and W chromosomes.

Bombyx mori is a female-heterogametic organism (female, ZW; male, ZZ) that appears to have a putative feminizing gene (Fem) on the W chromosome. The paternally transmitted mutant W chromosome, Df(p ( Sa ) + ( p )W + ( od ))Fem, derived from the translocation-carrying W chromosome (p ( Sa ) + ( p )W + ( od )), is inert as femaleness determinant. Moreover, this Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome has been thought to have a female-killing factor because no female larvae having the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome are produced. Initially, to investigate whether the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome contains any region of the W chromosome or not, we analyzed the presence or absence of 12 W-specific RAPD markers. The Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome contained 3 of 12 W-specific RAPD markers. These results strongly indicate that the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome contains the region of the W chromosome. Moreover, by using phenotypic and molecular markers, we confirmed that the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome is connected with a partially deleted Z chromosome and that this fused chromosome behaves as a Z chromosome during male meiosis. Furthermore, we demonstrated that the ZZW-type triploid female having the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome is viable. Therefore, we concluded that the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome does not have a female-killing factor but that partial deletion of the Z chromosome causes the death of the ZW-type diploid female having the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome. Additionally, our results of detailed genetic analyses strongly indicate that the female-killing chromosome composed of the Df(p ( Sa ) + ( p )W + ( od ))Fem chromosome and deleted Z chromosome was generated by translocation between the Z chromosome and the translocation-carrying W chromosome, p ( Sa ) + ( p )W + ( od ).

Clinical and physiological significance of abnormally prolonged central motor conduction time in HAM/TSP.

We measured the central motor conduction time (CMCT), central sensory conduction time (CSCT), F wave and mean F wave/M wave amplitude ratio in patients with HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP) and controls. CMCTs in upper (U) and lower (L) limbs were significantly prolonged in HAM/TSP. L-CSCT was significantly prolonged in HAM/TSP, but U-CSCT in HAM/TSP and controls were not significantly different. CMCT and CSCT were significantly correlated in HAM/TSP. U-CMCT, but not L-CMCT, correlated with the clinical severity of HAM/TSP. Although F wave conduction velocity and its occurrence were normal in HAM/TSP, U- and L-mean F wave/M wave amplitude ratio tended to be higher in HAM/TSP, and the L-mean F wave/M wave amplitude ratio was significantly correlated with the L- and thoracic CMCT. These findings demonstrate that the prolongation of CMCT sensitively reflects the extension of the lesions and the disinhibition to the anterior horn cells in HAM/TSP.

Two consecutive fasciculation potentials having different motor unit origins are an electromyographically pathognomonic finding of ALS.

To electrophysiologically diagnose amyotrophic lateral sclerosis (ALS), fasciculation potentials (FPs) were evaluated in each wasted muscle in 12 ALS and 14 other neurogenic disorders (non-ALS patients). Various types of FPs were observed in ALS. The number of discharged FPs and firing rate of FPs were significantly increased in ALS compared to those in non-ALS. These findings indicate that more motor units take part in discharging FPs in ALS than in non-ALS, and that injured lower motor neurons exhibit hyperexcitability. Although nine of 12 ALS patients showed two consecutive FPs having different motor unit origins, non-ALS patients did not show such FPs. Since the two consecutive FPs having different motor unit origins reflect an involvement of both upper and lower motor neurons, these potentials are an electrophysiologically pathognomonic finding to ALS.

Elevated plasma levels of transforming growth factor beta1 in patients with muscular dystrophy.

The progression of muscular weakness of patients suffering from muscular dystrophies directly correlates with the progressive loss of myofibers, accompanied by fibrosis. Since transforming growth factor beta1 (TGF-beta1) promotes tissue fibrosis, we measured the plasma TGF-beta1 level in patients with various muscular dystrophies in order to determine whether the level is elevated in patients with muscular dystrophy and if the level reflects the severity of tissue fibrosis. The plasma TGF-beta1 level was significantly elevated in patients with Duchenne muscular dystrophy and congenital muscular dystrophy (CMD), but not in those with Becker muscular dystrophy. Growth factors related to muscle fiber regeneration and fibrosis might be a key factor in the progression of muscular dystrophy and could be a target for therapeutic studies.

[Leigh syndrome: serial CT and MR imaging findings].

Serial CT and MRI findings in five patients (two boys and three girls) with Leigh syndrome were retrospectively reviewed in a follow-up period lasting from six months to 10 years. The two boys were found to have cytochrome c oxidase deficiency and one of the girls to have mitochondrial DNA mutation, while the remaining two girls had no detectable enzyme deficiency. CT and MRI revealed symmetrical involvement of the brain bilaterally in all cases. The focal lesions were found most frequently in putamina and caudate nuclei (four cases each), followed by thalami (three cases), globi pallidi, and midbrain (two cases each). In addition, diffuse white matter and/or cortical lesions were disclosed in three cases. MRI and CT at an early stage of the disease revealed swollen, symmetrical lesions which showed shrinkage in size accompanied by the adjacent brain atrophy on later images. Some lesions were detected only in the early stage and were not shown in later images. Thus, careful reading of sequential changes appears to be required for accurate diagnosis of Leigh syndrome.

[Event related potentials and reaction time on auditory discrimination tasks (2nd report)--a study by trial-to-trial analysis].

Latency variability for single trial in the 4 components (N100, P200, N200 and P300) of the auditory event related potentials (ERP) and the temporal relationship to the choice reaction time (RT) were investigated in 11 normal subjects. Identification of each latency on single trials was derived by using a correlational-template procedure. Results showed that the mean value of the single trial latency in the 4 components was close to the peak latency in the averaged waveform. Significantly larger coefficient of variation was obtained for the N100 component and RT, when compared to the other components. Higher correlation between single trial latency and RT was found for N200 and P300 components. The P300 component latency yielded a significant temporal effect across trials (p < 0.05) showing increases in its latency in the later trials, whereas none of the RT did. These results may indicate that the P300 component latency is influenced directly by subtle changes in cerebral activities during an initial stage of human cognitive function occurring in each trial, whereas RT is additionally influenced by greater functional fluctuation during the process of motor execution for making responses.

The auditory evoked middle-latency responses (MLRs): their normative variation and generators.

Auditory evoked middle-latency responses (MLRs) to clicks were recorded in 68 normal adults (ages from 21 to 59 year-old) at 16 scalp locations, all referred to a balanced non-cephalic reference. Both Na and Pa components were clearly demonstrated in all the subjects studied through the digital high-pass filtering. Both Na and Pa components were distributed dominantly over the frontal area in all subjects. The amplitudes of Na and Pa were significantly higher with binaural stimulation than with a monaural stimulation, and the existence of binaural interaction was also suggested. Binaural effect was more prominent in Pa than in Na component. When the data recorded from F3 and F4 electrodes were compared, most of F3/F4 amplitude ratios of Na and Pa were included between 0.7 and 1.5. The F3/F4 ratio should be useful indices for the diagnosis of excessive laterality. We also demonstrated gradual phase shifts of Na and Pa between the frontal and temporo-occipital area in about one-third of the records. This phenomenon strongly suggests that both Na and Pa components are derived from complex generators.

Cognitive impairment in Parkinson's disease assessed by visuomotor performance system and P300 potential.

Motor impairment, clinically defined as bradykinesia has been considered as resulting only from motor system problems. Combined analysis of cognitive function and dynamic characteristics was studied in 20 patients with Parkinson's disease by a simple aiming task on visuomotor performance system. The dynamic characteristics in a Parkinson's disease patient was evaluated by two parameters: decreased amplitude of the voluntary movement (= low gain constant) and delayed initiation of voluntary movement (= long reaction time). The visual event-related potential elicited in a target detection paradigm (P300 component) was recorded in 12 patients with Parkinson's disease. P300 latency was significantly prolonged in the patient group than in the normal control group (p less than 0.05). P300 has been shown to be intimately related to the cognitive process in the human brain and might well serve as a tool to monitor and evaluate the cognitive state in a clinical situation. The main cause of cognitive involvement in Parkinson's disease may include coexisting dementia and defective motivation. This type of cognitive disturbance may also serve partly as a cause of bradykinesia.

Auditory sound agnosia without aphasia following a right temporal lobe lesion.

A 55-year-old right-handed man showed inability to recognize the meaning of non-verbal sounds without impairment of language comprehension after a cerebrovascular accident. His auditory acuity was intact and no other sign of agnosia, apraxia or aphasia was detectable. His errors on a test of sound recognition were acoustic rather than semantic. Brain CT scan showed a small lesion in the posterior part of the right temporal lobe. This case suggests that auditory sound agnosia without language disorder can ensure a lesion confined to the right hemisphere, and that the deficit is discriminative rather than associative in nature.