Intragenic deletion in the gene encoding ubiquitin carboxy-terminal hydrolase in gad mice.

The gracile axonal dystrophy (gad) mouse is an autosomal recessive mutant that shows sensory ataxia at an early stage, followed by motor ataxia at a later stage. Pathologically, the mutant is characterized by 'dying-back' type axonal degeneration and formation of spheroid bodies in nerve terminals. Recent pathological observations have associated brain ageing and neurodegenerative diseases with progressive accumulation of ubiquitinated protein conjugates. In gad mice, accumulation of amyloid beta-protein and ubiquitin-positive deposits occur retrogradely along the sensory and motor nervous systems. We previously reported that the gad mutation was transmitted by a gene on chromosome 5 (refs 10,11). Here we find that the gad mutation is caused by an in-frame deletion including exons 7 and 8 of Uchl1, encoding the ubiquitin carboxy-terminal hydrolase (UCH) isozyme (Uch-l1) selectively expressed in the nervous system and testis. The gad allele encodes a truncated Uch-l1 lacking a segment of 42 amino acids containing a catalytic residue. As Uch-l1 is thought to stimulate protein degradation by generating free monomeric ubiquitin, the gad mutation appears to affect protein turnover. Our data suggest that altered function of the ubiquitin system directly causes neurodegeneration. The gad mouse provides a useful model for investigating human neurodegenerative disorders.

Pelvic organ dysfunction is more prevalent and severe in MSA-P compared to Parkinson's disease.

AIMS: It is usually difficult to distinguish between idiopathic Parkinson's disease (PD) and parkinsonian-type multiple system atrophy (MSA-P) in the early stage. However, it is important to make a careful early-stage diagnosis. Therefore, we determined whether an examination of pelvic organ dysfunction would be helpful to distinguish between PD and MSA-P. METHODS: We recruited 61 patients with PD and 54 patients with MSA-P who were examined at our neurology clinic. The mean ages of the patients with PD and MSA-P were 67 and 64 years, respectively. The mean disease duration of both groups was 3.2 years. We administered a questionnaire on pelvic organ dysfunction to the PD and MSA-P groups. The questionnaire had sections focusing on bladder, bowel, and sexual function. Dysfunction, as described in the responses, was evaluated as normal, mild (>once a month), moderate (>once a week), or severe (>once a day). The Mann-Whitney U-test was used for statistical analysis. RESULTS: Compared with the PD group, the prevalence and severity of pelvic dysfunction in the MSA-P group was significantly higher for urinary urgency (MSA-P 76%, PD 58%, P<0.05), retardation in initiating urination (79%, 48%, P<0.05), prolongation in urination (79%, 72%, P<0.05), and constipation (58%, 31%, P<0.05). The quality-of-life index among pelvic organ dysfunctions indicated that urinary and bowel function was significantly more impaired in the MSA-P group than in the PD group. CONCLUSIONS: Urinary urgency, retardation in initiating urination, prolongation in urination, and constipation are more prevalent and severe in MSA-P compared to PD.

Endoscope-assisted greater neurovascular palatal bundle release in cleft palatoplasty.

Performing surgery in the oral cavity is difficult because of the limited view of the surgical field. Intraoral surgery for infantile oral disorders, such as cleft palate, is even more challenging. Endoscopy provides a minimally invasive approach and clear surgical view in surgeries with a constrained field of view. To date, very few reports have described endoscope-assisted palate surgery for children with cleft palate. At the authors' institution, endoscopes have been used in primary palatoplasty using the double-opposing Z-plasty technique. A novel endoscope-assisted procedure is described herein, in which a dissection around the greater palatine neurovascular bundle is used to obtain tension-free closure of the palatal cleft. With this technique, it was possible to minimize the application of additional von Langenbeck-type relaxation incisions, which were previously introduced in most of our cases; the relaxation incision was successfully circumvented in 42.3% of cases. This led to lesser surgical interference, which possibly resulted in favourable palatal development. It was also found that the endoscopic procedure did not increase the operation time or blood loss when compared to those patients who underwent the non-endoscopic procedure. It is concluded that endoscopic guidance is quite useful in primary palatoplasty procedures with a constricted surgical view.

An uncommon cleft subtype of unilateral cleft lip and palate.

The finding that the vomer plays a crucial role in maxillary growth suggests that the bilateral cleft configuration of unilateral cleft lip and palate (UCLP), in which the vomer is detached from the non-cleft-side secondary hard palate, negatively influences palatal development, and this hypothesis was tested. Sixty persons with complete UCLP, including those with the vomer detached from (n = 30, b-UCLP) and attached to (n = 30, u-UCLP) the secondary hard palate, were analyzed morphologically, with the use of cast models taken at 10 days, 3 mos, and 12 mos of age. The anterio-posterior palatal length at 12 mos of age in those with b-UCLP was significantly shorter than that in those with u-UCLP, by 8.7% (p < 0.05). In addition, palatal width development in the first year in those with b-UCLP was also significantly retarded. These results suggest that the uncommon bilateral cleft subtype in UCLP should be included in the cleft classification.

Inhibition of trigeminal respiratory activity by suckling.

UNLABELLED: The trigeminal motor system is involved in many rhythmic oral-motor behaviors, such as suckling, mastication, swallowing, and breathing. Despite the obvious importance of functional coordination among these rhythmic activities, the system is not well-understood. In the present study, we examined the hypothesis that an interaction between suckling and breathing exists in the brainstem, by studying the respiratory activity in trigeminal motoneurons (TMNs) during fictive suckling using a neonatal rat in vitro brainstem preparation. The results showed that fictive suckling, which was neurochemically induced by bath application of N-methyl-D,L-aspartate and bicuculline-methiodide, or by local micro-injection of the same drugs to the trigeminal motor nucleus, inhibited the inspiratory activities in both respiration TMNs and respiratory rhythm-generating neurons. Under patch-clamp recording, fictive suckling caused membrane potential hyperpolarization of respiration TMNs. We conclude that the brainstem preparation contains an inhibitory circuit for respiratory activity in the trigeminal motor system via the rhythm-generating network for suckling. ABBREVIATIONS: BIC, bicuculline methiodide; GABA, gamma aminobutyric acid; NMA, N-methyl-D,L-aspartate; NMDA, N-methyl-D-aspartate; and TMN, trigeminal motoneuron.

Micturitional disturbance in pure autonomic failure.

We obtained micturitional histories and performed urodynamic studies in six patients with pure autonomic failure. All patients had urinary symptoms. Urodynamic studies showed postmicturition residuals in two, small bladder capacities in two, detrusor hyperreflexia in four, low bladder compliance in two, detrusor-external sphincter dyssynergia in one, neurogenic sphincter electromyography in three, and denervation supersensitivity of the bladder in two. Micturitional disturbance is a common feature in pure autonomic failure because of peripheral and central types of abnormalities.

Micturitional disturbance in patients with chronic inflammatory demyelinating polyneuropathy.

Eight of 32 patients (25%) with chronic inflammatory demyelinating polyneuropathy (CIDP) had micturitional disturbance, which consisted of voiding difficulty (n = 4), urgency (n = 4), or urgency incontinence (n = 1). Urodynamic studies on four symptomatic patients showed disturbed bladder sensation in two, bladder areflexia in one, and neurogenic changes of the external sphincter in one, indicative of peripheral parasympathetic and somatic nerve dysfunctions. Cystometry also showed detrusor overactivity in two patients but no evidence of CNS involvement, evidence that bladder overactivity occurs by probable pelvic nerve irritation.

Effects of subthalamic nucleus stimulation on the micturation reflex in cats.

High frequency stimulation (HFS) of the subthalamic nucleus (STN) has been performed to reverse motor dysfunction in severe parkinsonian patients. Recent studies suggested that neural circuitry in the basal ganglia might regulate micturition function as well. In 15 adult male cats under ketamine anesthesia, in which spontaneous isovolumetric micturition reflex had been generated, we performed electrical stimulation and extracellular single unit recording in the STN. Electrical stimulation applied in the STN elicited inhibition of the micturition reflex. None of the responses was facilitatory. Effective amplitude of the electrical stimulation for evoking inhibitory responses was less than 50 microA, which gradually increased and exceeded 250 microA as the location of the stimulation exceeded an area of the STN. Effective frequency of the electrical stimulation with given stimulus intensity was 50 Hz and higher. Total 10 neurons were recorded in the STN that were related to urinary storage/micturition cycles. All neurons were tonically active throughout storage/micturition cycles with storage phase predominance, with almost constant firing activities during the storage phase. In conclusion, our results showed that HFS-STN inhibited the micturition reflex and there were micturition-related neuronal firings in the STN in cats, suggesting the STN may be involved in neural control of micturition. The results also provide an implication that clinical HFS-STN may alter urinary function in parkinsonian patients.

An in-frame deletion in peripheral myelin protein-22 gene causes hypomyelination and cell death of the Schwann cells in the new Trembler mutant mice.

Cloning and sequencing of the peripheral myelin protein-22 cDNA and genomic DNA from newly found Trembler mice revealed an in-frame deletion including exon IV which codes for the second (TM2) and a part of third (TM3) transmembrane domain of peripheral myelin protein-22. This mutation was distinct from those in both other allelic Trembler and Trembler-J mice, which carry point mutations within the putative transmembrane spanning regions of peripheral myelin protein-22. Inheritance was autosomal dominant. The affected mice revealed an abnormal gait, which appeared at 15-20 days of age, followed by motor and sensory ataxia, which remained throughout life. Most of the affected mice could survive more than one year. One of the most notable pathological phenotypes was a giant vacuolar formation in the sciatic nerve of homozygotes. They vary in size within the cytoplasm of Schwann cells, which failed to assemble myelin at any ages studied. Heterozygotes showed normal myelination during the early postnatal stages, followed by a segmental demyelination at an advanced stage. Vacuolar formation was not so frequent as in the homozygotes. These results suggest that the missing of transmembrane spanning region (TM2 and TM3) of peripheral myelin protein-22 may disturb a dual biological function of peripheral myelin protein-22, leading to a dysmyelination of axons and to a vacuolar formation within the cytoplasm of the Schwann cells. The latter phenotype is discussed in conjunction with the disruption of an intracellular transport system and subsequent cell death.

The role of M(2)-muscarinic receptors in mediating contraction of the pig urinary bladder in vitro.

1. In urinary bladder, M(2)-muscarinic receptors predominate, but it is the smaller population of M(3)-receptors which mediate detrusor contraction. This study examines the M(2) : M(3) ratio and the role of M(2)-receptors in contraction of pig urinary bladder. 2. Competition experiments with [(3)H]-QNB determined the ratio of M(2) : M(3). In functional studies, affinity values (pK(B)) for 4-DAMP, darifenacin and methoctramine were calculated. Similar experiments were performed on tissues following selective M(3)-inactivation (incubation with 40 nM 4-DAMP mustard in the presence of 1 microM methoctramine to protect M(2)-receptors), precontraction with 50 mM KCl and relaxation with isoprenaline (30 microM) or forskolin (1 microM). 3. In competition binding, displacement of [(3)H]-QNB by 4-DAMP, darifenacin and methoctramine best fitted a two-site model suggesting a predominant (70 - 80%) population of M(2)-receptors. 4. On normal detrusor in vitro, 4-DAMP and methoctramine caused surmountable antagonism of responses to carbachol with pK(B) values of 9.37+/-0.07 and 6.05+/-0.05 respectively. Darifenacin caused unsurmountable antagonism, the apparent pK(B) value being 8.61+/-0.10. 5. In tissues where the M(3)-receptors had been inactivated and cyclic AMP levels elevated, 4-DAMP and darifenacin were less potent, with apparent pK(B) values of 8.72+/-0.08 and 6.74+/-0.07. In contrast, methoctramine was more potent, the apparent pK(B) value increasing significantly to 6.86+/-0.06. 6. se data suggest that the pig bladder possesses a similar muscarinic receptor population to the human bladder and that the M(3)-receptor subtype mediates contraction of the normal detrusor muscle. However an involvement of M(2)-receptors in contraction can be observed following pharmacological manipulation of the receptor population.

The influence of afferent inputs from skin and viscera on the activity of the bladder and the skeletal muscle surrounding the urethra in the rat.

(1) Somato-visceral and viscero-visceral reflex interactions have been studied in the bladder branches of the pelvic nerve and in the electromyographic (EMG) activity of the periurethral skeletal muscles of the anesthetized rat, and by observations of changes in bladder motility. (2) Slow distensions of the bladder caused some elevation of intravesical pressure, and culminated in a micturition contraction. Periurethral EMG activity increased gradually during the bladder distension, and showed an oscillatory marked increase during the bladder contraction. There was a small increase in pelvic nerve efferent activity during slow distension, and there was a substantial increase before, or at the start, of a micturition contraction. (3) Oscillatory bursting activity occurred in recordings of the EMG activity from periurethral skeletal muscle during the rising phase of micturition contraction; this was particularly so during the most rapid rise in intravesical pressure, and periods of electrical silence lasting 80-270 ms alternated with bursts of activity in the periurethral EMG. (4) In the present experiments, the switching mechanism activated by pelvic afferent signals related to intravesical pressure reversed the behavior of a number of reflex pathways. When the bladder pressure was low, nociceptive pinching of the perineal skin usually caused bladder contraction and a rise in pelvic nerve efferent activity and in periurethral EMG activity. When the bladder was full, micturition contractions were present and reduced in size and frequency by pinching of the perineal skin. The pelvic nerve efferent activity was correspondingly reduced, while the EMG activity increased during and following the nociceptive stimulus. Cooling the scrotal skin with ice also decreased the frequency of bladder contractions. (5) When the bladder pressure was low, distension of the anus and colon increased periurethral EMG activity, but did not affect bladder tone. However, when the bladder was full, these stimuli reduced the size and frequency of bladder contractions, associated with a reduction in the pelvic nerve efferent activity. There was usually a simultaneous reduction in the EMG activity in periurethral muscles. Similar results were obtained during distension of the seminal vesicles or vagina, or following injection of 20-60 microliters of saline into the lumen of the vas deferens. Reversal of the responses at extremes of intravesical pressure was observed in every case. (6) Following spinal transection at the upper cervical or thoracic level, micturition contractions were absent at high bladder volumes. However the effects described when the neuraxis was intact and the bladder pressure was low were still observed.(ABSTRACT TRUNCATED AT 400 WORDS)

Sequence analysis, tissue expression and chromosomal localization of a mouse secreted superoxide dismutase gene.

Using a reverse transcriptase polymerase chain reaction (RT-PCR), a complement DNA encoding secreted superoxide dismutase (s-SOD) of a mouse kidney has been isolated and the nucleotide sequence was determined. The deduced amino acid sequence of mouse s-SOD cDNA shares 79% identity with the rat seminal SOD sequence and 61% identity with the human SOD3 sequence. Northern blot analysis showed that mouse s-SOD is intensely expressed in the kidney and lung tissues and detectable in other tested tissues, including the brain. The mouse s-SOD gene was assigned to chromosome 5 using fluorescence in situ hybridization analysis and PCR analysis of mouse/hamster hybrid cells.

Antinociceptive synergism between supraspinal and spinal sites after subcutaneous morphine evidenced by CNS morphine content.

Morphine antinociception after various administration routes was estimated by the tail-flick method in rats. The antinociceptive ED50 (AD50) values for i.c.v., i.t., i.c.v. + i.t. (4:1 dose ratio) and s.c. were 6.9 micrograms, 0.49 + 0.12 micrograms and 2.7 mg/kg, respectively. Isobolographic analysis of AD50 (except s.c.) suggested that concurrent administration of i.c.v. and i.t. morphine interacted multiplicatively to produce antinociception. Morphine content in the CNS after administration of AD50 morphine for each route was estimated. Isobolographic analysis of morphine content revealed that supraspinal and spinal morphine interacted multiplicatively to produce antinociception after i.c.v. + i.t. and s.c. administration. Comparison of the dose-response curves (i.c.v. alone, i.t. alone, various i.c.v. + fixed i.t., fixed i.c.v. + various i.t.) suggested that supraspinal and spinal morphine can potentiate the antinociception induced by the other site, and that they have almost equal importance in the antinociceptive synergism. These results provide direct evidence for the synergism between supraspinal and spinal morphine to play an important role in the antinociception of systemically administered morphine.

Micturitional disturbance and the pontine tegmental lesion: urodynamic and MRI analyses of vascular cases.

Micturitional histories were taken from 39 patients with acute brainstem stroke. Within 3 months from onset, 49% had irritative as well as obstructive urinary symptoms, the most common being voiding difficulty and nocturnal urinary frequency in 28%, followed by urinary retention in 21%. Urodynamic studies of 11 symptomatic patients revealed detrusor hyperreflexia in 73%, low compliance bladder in 9%, atonic cystometrogram in 27%, detrusor-sphincter dyssynergia in 45% and uninhibited sphincter relaxation in 27%. Three asymptomatic patients had normal urodynamic findings. Brain magnetic resonance images of the lesions of the symptomatic patients were concentrated in the dorsolateral pons including pontine reticular nucleus and the reticular formation adjacent to the medial parabrachial nucleus and the locus coeruleus. These regions seem to be mainly responsible for supranuclear types of pelvic and pudendal nerve dysfunction in our patients with brainstem stroke, corresponding to the pontine urinary storage and micturation center reported in animal studies.

Micturitional disturbance after acute hemispheric stroke: analysis of the lesion site by CT and MRI.

Micturitional histories and urodynamic studies were performed in 72 acute hemispheric stroke patients. Within 3 months from the onset, 53% of the patients had urinary symptoms including irritative as well as obstructive, and the most common symptom was nocturnal urinary frequency in 36%, which was followed by urge urinary incontinence in 29% and difficulty of voiding in 25% of the patients. We found a correlation between micturitional disturbance with hemiparesis (p <0.05) and not with hemianopsia (p <0.05). Micturitional disturbance was more common in lesions of the frontal lobe (p <0.05) than in those of the occipital lobe. Brain CT or MRI in symptomatic patients showed lesions of anterior and medial surface of the frontal lobe, anterior edge of the paraventricular white matter, genu of the internal capsule and large lesions of putamen or thalamus. Urodynamic studies of 22 symptomatic patients revealed various findings in 91% of them, including detrusor hyperreflexia in 68%, detrusor-sphincter dyssynergia (DSD) in 14% and uninhibited sphincter relaxation in 36%. Patients with urinary retention had atonic cystometrogram and DSD. Detrusor hyperreflexia was noted in lesions of the frontal lobe as well as the basal ganglia, uninhibited sphincter relaxation in the frontal lobe, and detrusor-sphincter dyssynergia common in the basal ganglia. Above findings seem to indicate that anteromedial frontal lobe and its descending pathway, and the basal ganglia seem to be mainly responsible for supranuclear types of pelvic and pudendal nerve dysfunction in our patients with stroke.

Micturitional disturbance in syringomyelia.

Micturitional histories and urodynamic studies were performed in 14 patients with syringomyelia. Eleven patients were revealed to have urinary symptoms including difficulty of voiding in 8, urinary retention in 3, nocturnal and diurnal urinary frequency in 3, urinary incontinence in 2, and sense of urgency and enuresis in one. These urinary symptoms appeared after 5.3 years (ranging from 2 months to 13 years) from the occurrence of the neurological symptoms. Urodynamic studies revealed detrusor hyperreflexia in 7, detrusor areflexia in 4, detrusor-sphincter dyssynergia in 4 and uninhibited sphincter relaxation in 2 patients. Analysis of the motor unit potentials of the external sphincter revealed 5 of 6 patients had high amplitude or polyphasic neurogenic changes. Supranuclear as well as nuclear types of parasympathetic and somatic nerve dysfunctions seemed to be responsible for micturitional disturbance in our patients with syringomyelia. During the follow-up period of 2 to 63 months, urinary symptoms gradually improved in 4 of 6 patients after syringosubarachnoid shunts and in 3 of 4 after alpha-adrenoreceptor blocking agents.

SPECT imaging of the dopamine transporter with [(123)I]-beta-CIT reveals marked decline of nigrostriatal dopaminergic function in Parkinson's disease with urinary dysfunction.

We studied a correlation of urinary dysfunction with nigrostriatal dopaminergic deficit in Parkinson's disease (PD) by single-photon emission computed tomography (SPECT) imaging of dopamine transporter with [123I]-2beta-carbomethoxy-3beta-(4-iodophenyl)tropane (beta-CIT). Eleven patients were enrolled in the study, including four men and seven women, with a mean age of 64 years. Seven patients had urinary symptoms 1-5 years after the onset of motor disorder, which included nighttime frequency in six, urinary retardation in four, daytime frequency in one and urge urinary incontinence in one. Using a SPECT camera, the ratio specific to nondisplaceable [123I]-beta-CIT uptake, designated as "striatal V3" was obtained in the caudate, anterior and posterior putamen 24 h after the tracer injection. The striatal V3 was compared in patients with and without urinary dysfunction, and between men and women, using unpaired Student's t-test. Correlation of motor dysfunction and duration of illness with urinary dysfunction, was also analyzed. In the patients, there was a reduction of [123I]-beta-CIT binding in the striatum on both sides, particularly in the putamen contralateral to the affected body side. The striatal V3 of the caudate (p<0.01, Rt; p<0.05, Lt), anterior putamen (p<0.05, Rt) and posterior putamen (p<0.05, Rt) in patients with urinary dysfunction was significantly reduced than those without urinary dysfunction. No sex difference was seen in reduction of [123I]-beta-CIT binding. Urinary dysfunction in PD was more common in patients with higher Unified Parkinson's Disease Rating Scale (UPDRS) score, higher Hoehn-Yahr grade, but not in those with longer duration of disease, although there was no statistical significance. It is likely that our results reflect the association of urinary dysfunction and degeneration of the nigrostriatal dopaminergic cells in PD.

Breast-conservation treatment for patients with ductal carcinoma in situ.

Fourteen cases with symptomatic ductal carcinoma in situ (DCIS) were treated with breast-conservation treatment intensified with endocrine therapy. Nine of 14 patients with palpable mass had tumor detected on mammography. CT, ultrasonography, and MRI were able to detect linear and/or spotty lesion or enhancement suggesting DCIS. Whereas these findings were not particular to DCIS, the combination of these modalities would be useful in deciding the extent of resection for DCIS. There was no patient selection for breast-conservation treatment in our department. All patients received tangential and boost radiation, and were treated with endocrine therapy using anti-estrogen drugs. The reason that nine cases had close margins (<5 mm) might be on account of the treatment including lumpectomy with 1 cm of surgical margin. In spite of their margin status, no local or systemic failure was experienced, and the cosmetic results of most patients were rated as excellent or good. Therefore, our breast-conservation treatment intensified with systemic therapy is thought to be adequate for patients with symptomatic DCIS. Six of eight cases who received preoperative treatment containing endocrine therapy with or without CAF chemotherapy showed a decrease in tumor size. Preoperative treatment may effect the microinvasion and/or breast tissue surrounding a DCIS tumor.

Adrenalectomy-induced potentiation of morphine analgesia: reversal by prednisolone.

Effects of adrenalectomy (ADX) on analgesic potency and morphine (MOR) content after SC administration of 3.5 or 7 mg/kg of MOR, and effects of prednisolone (PRED) on the ADX-induced effects were studied. ADX significantly potentiated MOR analgesia at both MOR doses, and PRED reversed the ADX-induced potentiation of MOR analgesia, ADX did not affect MOR content in brain and plasma after 3.5 mg/kg MOR, but significantly increased MOR content in brain and plasma after 7 mg/kg MOR, and PRED reversed the ADX-induced increase in the MOR content. Although the analgesic potency of 3.5 mg/kg MOR in ADX group was equipotent with those of 7 mg/kg MOR in sham-operated and PRED-treated ADX groups, MOR content in the former group was significantly lower than those in the latter two groups. These results suggest that ADX potentiates MOR analgesia through both mechanisms of the increased MOR content and the increased sensitivity to MOR, and that the lack of glucocorticoids participates in both of these ADX-induced effects.