Morphology of the cockerel's comb after androgen administration.

1. Androgen receptor (AR) expression and morphological changes in blood capillaries were investigated in the comb of cockerels, both untreated controls and after the administration of testosterone (T) or the androgen antagonist flutamide (F) for 7 weeks. 2. Twenty-six male Single Comb White Leghorn roosters were divided into T-treated, F-treated and untreated groups. Tissue sections were examined histologically, immunohistochemically, and comb blood vessel castings were analysed by scanning electron microscopy. 3. Histologically, the capillaries of the peripheral dermis layer in the T-treated group were dilated compared with controls. Many red blood cells were seen in the lumen. Although the capillary diameter in the F-treated group did not show a significant difference as compared with control, blood capillaries with small diameters were often observed, and there were few red blood cells in the capillaries. Some capillary castings were extended markedly in the T treated group, and small blood vessels were observed arborising from the extended blood capillaries. In contrast, all capillaries were slender in the F-treated group, and the casting surface was rough. 4. Immunoreactivity for AR was found in capillary endothelial cells in the peripheral dermis layer of the comb. The intensity of staining in these cells was increased in the T-treated group but was reduced in the F-treated group. 5. It is concluded that immunoreactivity for AR was found in capillary endothelial cells in the peripheral dermis layer of the rooster's comb. The intensity of staining in these cells was increased in the T-treated group but reduced in the F-treated group. Thus, the capillary endothelial cells in the peripheral dermis layer of the comb are androgen targets.

Treatment of a citrin-deficient patient at the early stage of adult-onset type II citrullinaemia with arginine and sodium pyruvate.

Citrin deficiency is a common congenital metabolic defect not only in East Asian populations but also in other populations around the world. It has been shown that although liver transplantation is ultimately required in many patients to prevent neurological decompensation associated with hyperammonaemia, arginine is effective in lowering ammonia in hyperammonaemic patients, and a high-protein low-carbohydrate diet may provide some benefit to infants in improving failure to thrive. In the present study, the clinical symptoms and laboratory findings are reported for a 13-year-old citrin-deficient girl in the early stage of adult-onset type II citrullinaemia (CTLN2), and the therapeutic effect of orally administered arginine and sodium pyruvate was investigated. The patient complained of anorexia, lethargy, fatigue and poor growth, and showed laboratory findings typical of CTLN2; elevated levels of plasma citrulline, threonine-to-serine ratio, and serum pancreatic secretory trypsin inhibitor. Oral administration of arginine and sodium pyruvate for over 3 years improved her clinical symptoms and has almost completely normalized her laboratory findings. It is suggested that the administration of arginine and sodium pyruvate with low-carbohydrate meals may be an effective therapy in patients with citrin deficiency in order either to prolong metabolic normalcy or to provide a safer and more affordable alternative to liver transplantation.

Reduced carbohydrate intake in citrin-deficient subjects.

Citrin is the liver-type aspartate-glutamate carrier that resides within the inner mitochondrial membrane. Citrin deficiency (due to homozygous or compound heterozygous mutations in the gene SLC25A13) causes both adult-onset type II citrullinaemia (CTLN2) and neonatal intrahepatic cholestasis (NICCD). Clinically, CTLN2 is characterized by hyperammonaemia and citrullinaemia, whereas NICCD has a much more varied and transient presentation that can include multiple aminoacidaemias, hypoproteinaemia, galactosaemia, hypoglycaemia, and jaundice. Personal histories from CTLN2 patients have repeatedly described an aversion to carbohydrate-rich foods, and clinical observations of dietary and therapeutic outcomes have suggested that their unusual food preferences may be directly related to their pathophysiology. In the present study, we monitored the food intake of 18 Japanese citrin-deficient subjects whose ages ranged from 1 to 33 years, comparing them against published values for the general Japanese population. Our survey confirmed a marked decrease in carbohydrate intake, which accounts for a smaller proportion of carbohydrates contributing to the total energy intake (PFC ratio) as well as a shift towards a lower centile distribution for carbohydrate intake relative to age- and sex-matched controls. These results strongly support an avoidance of carbohydrate-rich foods by citrin-deficient patients that may lead to worsening of symptoms.

High prevalence of bipolar disorder comorbidity in adolescents and young adults with high-functioning autism spectrum disorder: a preliminary study of 44 outpatients.

BACKGROUND: Psychiatric comorbidity of autism spectrum disorder (ASD) has not been well examined. METHODS: Mood disorders in 44 consecutive outpatients with high-functioning ASD were examined at a university hospital according to DSM-IV. Inclusion criteria were an IQ of 70 or higher on the Wechsler Intelligence Scale and age of 12 years or over. RESULTS: Sixteen patients (36.4%) were diagnosed with mood disorder. Of these 16 patients, four were diagnosed as having major depressive disorder, two patients as bipolar I disorder, six patients as bipolar II disorder, and four patients as bipolar disorder not otherwise specified. Bipolar disorder accounted for 75% of cases. Twelve patients had Asperger disorder and four patients had pervasive developmental disorder not otherwise specified. None of the patients had autistic disorder. LIMITATIONS: The sample size was small. We could not use Autism Diagnostic Interview - Revised. Referral bias could not be avoided in this study. CONCLUSIONS: The major comorbid mood disorder in patients with high-functioning ASD is bipolar disorder and not major depressive disorder. The autistic spectrum may share common vulnerability genes with the bipolar spectrum.

Ghrelin stimulates phagocytosis and superoxide production in fish leukocytes.

To clarify the role of ghrelin in the fish immune system, the in vitro effect of ghrelin was examined in phagocytic leukocytes of rainbow trout (Oncorhynchus mykiss). Administration of trout ghrelin and des-VRQ-trout ghrelin, in which three amino acids are deleted from trout ghrelin, increased superoxide production in zymosan-stimulated phagocytic leukocytes from the head kidney. Gene expression of growth hormone (GH) secretagogue-receptor (GHS-R) was detected by RT-PCR in leukocytes. Pretreatment of phagocytic leukocytes with a GHS-R antagonist, [D-Lys3]-GHRP-6, abolished the stimulatory effects of trout ghrelin and des-VRQ-trout ghrelin on superoxide production. Ghrelin increased mRNA levels of superoxide dismutase and GH expressed in trout phagocytic leukocytes. Immunoneutralization of GH by addition of anti-salmon GH serum to the medium blocked the stimulatory effect of ghrelin on superoxide production. These results suggest that ghrelin stimulates phagocytosis in fish leukocytes through a GHS-R-dependent pathway, and also that the effect of ghrelin is mediated, at least in part, by GH secreted by leukocytes.

Non-Menkes-type copper deficiency with regression, lactic acidosis, and granulocytopenia.

A 2-year-old girl with granulocytopenia developed fever followed by truncal ataxia and progressive neurologic regression. CT demonstrated symmetric low-density areas in the cerebral white matter. Sural nerve biopsy revealed axonal degeneration. Blood lactate levels were high, and serum levels of copper and ceruloplasmin and urinary excretion of copper were low. Cultured skin fibroblasts showed normal copper uptake. Treatment with oral copper administration normalized serum copper and ceruloplasmin levels, blood lactate levels, and granulocyte count. However, copper levels in the CSF were still low, and the patient showed no clinical improvement. We speculated that copper transport across the intestinal wall and across the blood-brain barrier was impaired.

Prediction of the development of atopic symptoms in early childhood by cord IgE-binding factors (soluble Fc epsilon R2).

The relationship of cord serum sFc epsilon R2 levels to the development of atopic symptoms in early childhood was studied. Cord sFc epsilon R2 was 444.2 +/- 235.1 pg/ml (n = 77), which was not significantly different from maternal serum sFc epsilon R2 (541.7 +/- 346.9 pg/ml, n = 42). However, there was no correlation between cord and maternal serum sFc epsilon R2, suggesting that most, if not all, of cord serum sFc epsilon R2 was produced by the fetus itself. Cord serum sFc epsilon R2 in infants who developed atopic symptoms later was significantly higher than that in infants who were free of atopic symptoms (P less than 0.01 at 7 and 13 months of age). The incidence of the development of atopic symptoms increased with the increase of cord serum sFc epsilon R2. These results suggest that sFc epsilon R2 is related to the development of atopic disorders and that the measurement of cord serum sFc epsilon R2 may be of value in predicting the development of atopic disorders in early childhood.

Involvement of the noradrenergic system in the seizures of epileptic El mice.

We studied the role of the noradrenergic system in the seizures of epileptic El mice. To this end, the anticonvulsant activity of adrenergic drugs was tested with a scoring method, and the binding of [3H]dihydroalprenolol, [3H]prazosin and [3H]yohimbine was evaluated in whole brains and various brain regions from stimulated and unstimulated El mice, and their maternal ddy mice. The seizures of El mice were inhibited by noradrenaline, phenylephrine, oxymetazoline, clonidine and yohimbine in a dose-dependent manner. These preventive effects of alpha-adrenoceptor agonists were antagonized by pretreatment with alpha-adrenoceptor antagonists. The preventive effect of yohimbine was reversed by pretreatment with clonidine or alpha-methyl-p-tyrosine, although the latter drug did not affect the anticonvulsant effect of clonidine. The binding of [3H]dihydroalprenolol was the same in the three groups of mice. More [3H]prazosin was bound in the cerebellum and striatum, and there were more [3H]yohimbine binding sites in the whole brain, cerebral cortex, hippocampus and brainstem of stimulated and unstimulated El mice than in the same areas of ddy mice. These findings suggest that up-regulated alpha 1- and alpha 2-adrenoceptors are involved in the inhibition of the seizures of El mice.

MELAS of infantile onset: mitochondrial angiopathy or cytopathy?

An 83-day-old male infant had convulsions, hypertrophic cardiomyopathy, and lactic acidosis. Cranial computed tomography revealed low-density areas in both parieto-occipital lobes and in the left temporal lobe. Muscle biopsy did not reveal ragged-red fibers, but abnormal mitochondria were found in the capillary endothelial cells as well as in the muscle fibers. At 5 months of age, the patient developed purpura on the soles and palms. Skin biopsy showed degeneration of the endothelial cells with abnormal mitochondria in the arterioles and capillaries. Myelinated nerves in the skin had vacuolated axons with swollen mitochondria, and their myelin sheaths showed vacuolation. At 9 months of age, he died of heart failure, and autopsy revealed abnormal mitochondria in the myocardium but not in the coronary vessels. Our findings indicate that the symptoms of the mitochondrial encephalopathy, myopathy, lactic acidosis, and strokelike episodes (MELAS) syndrome cannot be fully explained by the mitochondrial angiopathy alone.

Association of NMDA receptor sites and seizures of El mice.

To investigate the possible role of N-methyl-D-aspartate (NMDA) receptors in the seizures of El mice, a genetic animal model of epilepsy, we measured [3H]3-[(+)-2-(carboxypiperazin-4-yl)][1,2-3H]- propyl-1-phosphonic acid (CPP) binding in several brain regions of El and ddY mice. At 22-24 weeks of age, the maximum number of binding sites (Bmax) of [3H]CPP was lower only in the cerebral cortex of both stimulated and unstimulated El mice (El(+) and El(-), respectively) than in that of ddY mice. A reduction in Bmax values of cortical [3H]CPP binding of El mice was detected after the age of 12 weeks. Cortical [3H]CPP binding in El(+) mice decreased further transiently after evoked seizures. No significant change was observed in El(-) mice after postural stimulation. These results suggest that El(+) and El(-) mice share seizure propensity and that activation of NMDA receptors is involved in the seizures of El mice.

NMDA-sensitive L-[3H]glutamate binding in cerebral cortex of El mice.

NMDA-sensitive L-[3H]glutamate binding was examined in the brains of El mice, a genetic animal model of epilepsy, and in ddY mice. In whole brain, Scatchard analysis showed that both stimulated and unstimulated El mice had significantly lower Bmax values for binding than did ddY mice. In regional studies, the binding of NMDA-sensitive L-[3H]glutamate was significantly less in the cerebral cortex of both stimulated and unstimulated El mice than in that of ddY mice. These data suggest that NMDA receptors may be involved in the genetic susceptibility of El mice to seizures.

Onset age-dependent variations of three islet specific autoantibodies in Japanese IDDM patients.

The age related incidence rate of insulin-dependent diabetes mellitus shows a bimodal distribution, not only in Caucasians but also in Japanese. To evaluate the onset age-related autoimmune profile at presentation in insulin-dependent diabetes mellitus (IDDM), glutamic acid decarboxylase (GAD) autoantibody, islet cell antibody (ICA), and insulin autoantibody (IAA) were measured in 137 newly diagnosed Japanese IDDM patients with onset ages between 0-29 years. The prevalence of GAD autoantibody was significantly increased from the lowest (32%) in the 0-5 years onset age group to 75% in the 13-19 years onset age group (P < 0.05), whereas the IAA prevalence significantly decreased from the peak (48%) in the 6-12 years onset age group to 10% in the 20-29 years onset age group (P < 0.05). The ICA prevalence was increased from the lowest (32%) in the 0-5 years onset age group to the highest (53%) in the 20-29 years onset age group similar to that for the GAD autoantibody. Such results demonstrate that there was age-related autoimmune characteristics at presentation of IDDM in Japanese as well as in Caucasians.

CT appearance of children in a persistent vegetative state.

A morphometric CT study was performed on 17 children who were in a persistent vegetative state. Four cases with compromised brain stem function (group 1) showed a significantly smaller lateral pontine ratio (LPR, width of pons/greatest internal diameter between temporal bone) than the remaining 13 cases with preserved brain stem function (group 2) and controls. LPR was considered a useful indicator of brain stem atrophy and to correlate well with brain stem function.

Regional cerebral blood flow in diagnosis of childhood onset partial epilepsy.

We compared regional cerebral blood flow assessed by [123I]N-isopropyl-p-iodoamphetamine (IMP) single-photon emission tomography (SPECT) with magnetic resonance imaging (MRI), computed tomography (CT) and interictal surface electroencephalography (EEG) to evaluate its diagnostic potential in 24 patients with partial seizures with onset in childhood. Focal low uptake areas were observed on SPECT scans of 18 patients and were presumed to represent epileptogenic areas in 17. MRI revealed an abnormality in 12 and CT in 6 patients, and all organic lesions showed SPECT abnormalities, too. Six patients without focal structural abnormalities showed regional perfusion deficit on SPECT. Routine scalp EEG revealed an epileptic focus in 17 patients and three of them showed discordant results between SPECT and EEG, which suggested more serious brain disorders. In two patients without EEG localization only SPECT showed focal abnormalities in the probable epileptic area. [123I]IMP-SPECT was useful in locating the epileptic focus, particularly during the early period after the onset of partial seizures when the EEG gave inconclusive results.

123I-IMP SPECT findings in mitochondrial encephalomyopathies.

We performed N-isopropyl-[123I]p-iodoamphetamine (IMP) single photon emission computed tomography (SPECT) in three patients with Leigh syndrome, two patients with mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS), and two siblings with progressive external ophthalmoplegia (PEO). The SPECT images were compared with the findings on magnetic resonance imaging (MRI) and computed tomography (CT). All Leigh syndrome patients showed low accumulation areas (LAA) bilaterally in the frontal lobes and the basal ganglia. The frontal lobe LAA was seen even in an area without abnormalities on CT/MRI. Each MELAS patient showed a focal LAA. SPECT could also detect an old stroke-like lesion that was no longer shown by CT/MRI. However, SPECT did not show LAA in the basal ganglia, which showed calcification on CT or abnormal signal intensity on MRI. MRI in the 2 PEO patients showed lesions bilaterally in the basal ganglia in one, and in the internal capsules in the other. SPECT showed LAA not only in corresponding areas, but also in the occipital lobes, where no lesions were revealed by MRI. Thus, 123I-IMP SPECT was more sensitive than CT/MRI for detecting stroke-like lesions in MELAS patients, although it did not detect small lesions in the basal ganglia. LAA in the frontal lobes and occipital lobes may be SPECT findings characteristic of Leigh syndrome and PEO, respectively.

Characteristics of auditory P300 in children: application of single trial analysis.

As, especially in children, cognitive function might vary from one stimulus to the other, even when the same target stimuli are presented in the oddball paradigm, we applied the single trial analysis adopting discrete Fourier transformation followed by inverse discrete Fourier transformation to the auditory P300 for the first time in children. In 9 children, age 9-13 years, a positive peak was clearly identified in approximately a quarter of the processed single sweep responses to target stimuli (single sweep P300), whereas it was observed less frequently for non-target stimuli. In 2 children of age 6 years, it was difficult to discriminate single sweep P300 from background EEG activities even in the processed data. As for the former group, the detection ratio of P300 in single sweep target responses ranged from 10.0 to 40.0%, and its mean latency and amplitude ranged from 311 to 348 ms and from 31.7 to 41.5 microV, respectively. There was a significant correlation between average data and single sweep data in terms of the P300 latency. However, the amplitude of P300 in the average data bore no relation to the detection ratio, mean amplitude or variability of latency among single sweep P300s. As compared with adult data previously reported, the detection ratio of single sweep P300 was lower and the P300 amplitude was higher in childhood. P300 in children, at least past puberty, can be analyzed by using the single trial method, which is expected to provide more information about the development of human higher brain functions, especially cognitive function.

Maturation of somatosensory evoked potentials upon posterior tibial nerve stimulation.

Somatosensory evoked potentials produced in response to posterior tibial nerve stimulation were studied in 42 normal infants and children, ages 4 months to 16 years. The maturation of afferent conduction from the lower limb was evaluated for the peripheral nerve, spinal cord, and central nervous system. Although the maturation of conduction in the peripheral nerve (from the ankle to the popliteal fossa and from the popliteal fossa to L3) was complete by 6 years of age, afferent conduction in the spinal cord (from L3 to C7) was not complete until 12 years of age or older. Spinal evoked potentials investigated in the thoracolumbar area revealed a phase-reversed potential located between the lower thoracic spine and upper lumbar spine in over 80% of patients. Reciprocal velocities for the major cortical positive potential P1 (corresponding to P37 in adults) and its onset, N1, steadily decreased with age and leveled off at greater than 12 years of age and by 12 years of age, respectively. The propagation velocity from L3 to the cerebral cortex also increased steadily with age, leveling off at greater than 12 years of age. Accordingly, the maturation of afferent conduction in the central nervous system was not complete until after 12 years of age.

Possible induction of systemic lupus erythematosus by zonisamide.

An epileptic male to whom zonisamide and ethosuximide had been administered exhibited fever, pericarditis, pleuritis, arthralgia, and elevated titers of antinuclear antibody and anti-DNA-antibody at 5 years, 11 months of age. It was suggested that zonisamide induced the systemic lupus erythematosus symptoms because the antinuclear antibody titers began to rise after zonisamide was administered, and clinical recovery and a decrease in the anti-DNA-antibody levels promptly followed the discontinuation of the drug. The positive lymphocyte transformation test results against zonisamide further supports the etiologic relationship of the male's lupus and the drug. This first case of zonisamide-induced lupus should alert physicians to the possibility of this serious unwanted effect in patients receiving zonisamide.

Somatosensory evoked potentials after posterior tibial nerve stimulation in focal spinal cord diseases.

We investigated the somatosensory evoked potentials (SEPs) produced by posterior tibial nerve (PTN) stimulation in 8 infants and children with focal spinal cord disorders. The spinal responses of the PTN-SEPs were considered to assist in the localization of spinal lesions because their abnormalities were consistent with the neurologic and/or radiologic findings in all 6 examinations that revealed abnormal spinal SEPs. The cortical responses correlated significantly with proprioception in the lower limbs because proprioception was only disturbed when the cortical SEPs were absent (3 examinations). When both the spinal and cortical responses were abnormal, the spinal lesion probably involved the dorsal column so extensively that it completely interrupted the afferent impulses. In contrast, when SEP studies demonstrated abnormal spinal and normal cortical SEPs (3 examinations), the dorsal column involvement was probably less severe; therefore, both the spinal and cortical responses provided useful information regarding afferent conduction in the dorsal column. PTN-SEPs appear to have the potential to be of value in the diagnosis of focal spinal disease, especially in infants and young children who cannot cooperate with detailed neurologic examinations.

Nontraumatic spinal intramedullary hemorrhage in an infant.

After an uncomplicated pregnancy and delivery, this female child suddenly became quadriplegic on the fifth day of life. She gradually regained movement in her upper extremities. At 2 months of age, she exhibited paraplegia with exaggerated deep tendon reflexes in the lower extremities. Babinski reflex was present bilaterally and sensory disturbances below the trunk were suspected. Somatosensory evoked potentials after median and ulnar nerve stimulation revealed preserved conduction from the upper extremities through the cervical spinal cord to the cortex. Somatosensory evoked potentials after posterior tibial nerve stimulation suggested disturbed conduction in the upper thoracic spinal cord. Magnetic resonance imaging disclosed a hypodense area in the thoracic cord between T1 and T4 on both the T1-weighted and gradient echo images consonant with an old hematoma cavity. Digital subtraction angiography failed to demonstrate any vascular malformation.