Compound heterozygous ADAMTS9 variants in Joubert syndrome-related disorders without renal manifestation.

BACKGROUND: Ciliopathies are the outcomes of defects of primary cilia structures and functions which cause multisystemic developmental disorders, such as polycystic kidney disease, nephronophthisis, retinitis pigmentosa, Joubert syndrome (JS), and JS-related disorders (JSRD) with additional organ involvement including oral-facial-digital syndrome and so on. They often share common and unexpected phenotypic features. CASE PRESENTATION: We report a 4-year-old-boy case with compound heterozygous variants of ADAMTS9. Unlike the cases with ADAMTS9 variants in the previous report, which identified that homozygous variants of ADAMTS9 were responsible for nephronophthisis-related ciliopathies in two cases, the current case did not have nephronophthisis nor renal dysfunction, and his clinical features, such as oculomotor apraxia, hypotonia, developmental delay, bifid tongue, and mild hypoplasia of cerebellar vermis indicated JSRD. CONCLUSIONS: The case suggested a possible association between the clinical presentation of JSRD and ADAMTS9-related disease, and it shows a wide spectrum of ADAMTS9 phenotype.

Comprehensive genetic analysis confers high diagnostic yield in 16 Japanese patients with corpus callosum anomalies.

Corpus callosum anomalies (CCA) is a common congenital brain anomaly with various etiologies. Although one of the most important etiologies is genetic factors, the genetic background of CCA is heterogenous and diverse types of variants are likely to be causative. In this study, we analyzed 16 Japanese patients with corpus callosum anomalies to delineate clinical features and the genetic background of CCAs. We observed the common phenotypes accompanied by CCAs: intellectual disability (100%), motor developmental delay (93.8%), seizures (60%), and facial dysmorphisms (50%). Brain magnetic resonance imaging showed colpocephaly (enlarged posterior horn of the lateral ventricles, 84.6%) and enlarged supracerebellar cistern (41.7%). Whole exome sequencing revealed genetic alterations in 9 of the 16 patients (56.3%), including 8 de novo alterations (2 copy number variants and variants in ARID1B, CDK8, HIVEP2, and TCF4) and a recessive variant of TBCK. De novo ARID1B variants were identified in three unrelated individuals, suggesting that ARID1B variants are major genetic causes of CCAs. A de novo TCF4 variant and somatic mosaic deletion at 18q21.31-qter encompassing TCF4 suggest an association of TCF4 abnormalities with CCAs. This study, which analyzes CCA patients usung whole exome sequencing, demonstrates that comprehensive genetic analysis would be useful for investigating various causal variants of CCAs.

A female patient with a hot spot mutation of PRRT2 gene suffering from several types of epileptic seizures in infancy.

Benign familial infantile epilepsy (BFIE) is characterized by non-febrile focal seizures, which sometimes evolve to secondarily generalized seizures and are usually resolved in the second year. Proline-rich transmembrane protein 2 (PRRT2) is confirmed as the major cause of BFIE, familial paroxysmal kinesigeneic dystonia (PKD) and infantile convulsions and choreoathetosis (ICCA) syndrome. We examined a female patient with a hot spot mutation of PRRT2 gene. She had recurrent tonic seizures when she was three months old. The seizures were controlled by several kinds of anticonvulsants. Then, she had several times of focal seizures daily at nine months old. However, the seizures were stopped by small amounts of carbamazepine. Later, when she was two years old, she experienced frequent motor seizures characterized by truncal flexion and swaying the body with partially disturbed consciousness. Her father also had the same PRRT2 gene mutation and non-febrile seizures in infancy. The patient had mild to moderate mental retardation, whereas her father was mentally normal. Therefore, the patient revealed a quiet different phenotype from that of her father as a carrier of the same PRRT2 gene mutation. We speculate that the PRRT2 mutation had caused the BFIE-like seizures both in the patient and her father, whereas other unknown genetic factors specific for the patient might be associated with the atypical seizures observed only in her.

Athetotic and spastic cerebral palsy: anatomic characterization based on diffusion-tensor imaging.

PURPOSE: To evaluate the anatomy of deep gray and white matter structures in children with athetotic cerebral palsy (CP) and those with spastic CP by using diffusion-tensor (DT) imaging and to investigate whether these types of CP have unique anatomic correlates that can support their diagnosis and prognosis. MATERIALS AND METHODS: This study was approved by the institutional review board of each participating institution, and written informed consent was obtained from the parents of each patient. DT imaging was used to retrospectively evaluate 19 children with clinically diagnosed athetotic CP (mean age, 3.4 years ± 3.3 [standard deviation]), 26 children with spastic CP (mean age, 3.3 years ± 3.2), and 31 healthy control subjects (mean age, 3.2 years ± 3.0). Fractional anisotropy (FA) and mean diffusivity (MD) were measured with a region of interest (ROI) method. The ROIs were drawn on bilateral deep gray and white matter structures, including projection fibers, association fibers, and commissural fibers. Statistical analysis was performed by using the Kruskal-Wallis test with Bonferroni correction. P < .05 indicated a significant difference. RESULTS: FA values in the athetotic CP group were significantly lower than those in the control and spastic CP groups for multiple structures, including deep gray and white matter (P < .05 and P = .0001, respectively); these differences were also associated with increasing MD (P < .05 and P < .001, respectively). On the other hand, in the spastic CP group, the significantly decreased FA values, compared with those of the normal group, were limited to several white matter structures (P < .05 and P = .0001). CONCLUSION: In children with athetotic CP, the extent of change on DT images due to early brain damage tends to be more diffuse, including multiple brain structures, compared with the changes in children with spastic CP.

Melanin-concentrating hormone 1-receptor antagonist suppresses body weight gain correlated with high receptor occupancy levels in diet-induced obesity mice.

Melanin-concentrating hormone (MCH), which is a neuropeptide expressed in the hypothalamus of the brain, is involved in regulating feeding behavior and energy homeostasis via the MCH(1) receptor in rodents. It is widely considered that MCH(1) receptor antagonists are worthy of development for medical treatment of obesity. Here we report on the development of an ex vivo receptor occupancy assay using a new radiolabeled MCH(1) receptor antagonist, [(35)S]-compound D. An MCH(1) receptor antagonist inhibited the binding of [(35)S]-compound D to brain slices in a dose-dependent manner. The result showed a good correlation between the receptor occupancy levels and plasma or brain levels of the MCH(1) receptor antagonist, suggesting that the ex vivo receptor binding assay using this radioligand is practical. Quantitative analysis in diet-induced obese mice showed that the efficacy of body weight reduction correlated with the receptor occupancy levels at 24h. Furthermore, more than 90% occupancy levels of MCH(1) receptor antagonists during 24h post-dosing are required for potent efficacy on body weight reduction. The present occupancy assay could be a useful pharmacodynamic marker to quantitatively estimate anti-obese efficacy, and would accelerate the development of MCH(1) receptor antagonists for treatment of obesity.

Discovery of novel diarylketoxime derivatives as selective and orally active melanin-concentrating hormone 1 receptor antagonists.

Optimization of the lead 2a led to the identification of a novel diarylketoxime class of melanin-concentrating hormone 1 receptor (MCH-1R) antagonists. Our focus was directed toward improvement of hERG activity and metabolic stability. The representative derivative 4b showed potent and dose-dependent body weight reduction in diet-induced obese (DIO) C57BL/6J mice after oral administration. The synthesis and structure-activity relationships of the novel diarylketoxime MCH-1R antagonists are described.

Discovery of pyridone-containing imidazolines as potent and selective inhibitors of neuropeptide Y Y5 receptor.

A series of 2-pyridone-containing imidazoline derivatives was synthesized and evaluated as neuropeptide Y Y5 receptor antagonists. Optimization of the 2-pyridone structure on the 2-position of the imidazoline ring led to identification of 1-(difluoromethyl)-5-[(4S,5S)-4-(4-fluorophenyl)-4-(6-fluoropyridin-3-yl)-5-methyl-4,5-dihydro-1H-imidazol-2-yl]pyridin-2(1H)-one (7m). Compound 7m displayed statistically significant inhibition of food intake in an agonist-induced food intake model in SD rats and no adverse cardiovascular effects in anesthetized dogs. In addition, markedly higher brain penetrability and a lower plasma Occ90 value were observed in P-gp-deficient mdr1a (-/-) mice compared to mdr1a (+/+) mice after oral administration of 7m.

Synergistic interaction between neuropeptide Y1 and Y5 receptor pathways in regulation of energy homeostasis.

Neuropeptide Y plays a key role in the physiological control of energy homeostasis. Five neuropeptide Y receptor subtypes have been cloned, and multiple neuropeptide Y receptor subtypes are thought to mediate neuropeptide Y activity. However, interactions among neuropeptide Y receptor subtypes have not been elucidated to date. Herein, we examined the interaction between neuropeptide Y(1) and Y(5) receptors in feeding regulation by employing selective neuropeptide Y(1) and Y(5) receptor antagonists in C57BL/6 and neuropeptide Y(1) receptor knockout mice fed a high-fat diet. A single-dose of a neuropeptide Y(1) receptor antagonist (10-30 mg/kg) suppressed spontaneous food intake and reduced body weight in high-fat diet-fed C57BL/6 mice, while treatment with a neuropeptide Y(5) receptor antagonist did not significantly reduce food intake or body weight. Coadministration of a neuropeptide Y(1) receptor antagonist with a neuropeptide Y(5) receptor antagonist further suppressed food intake and reduced body weight. Next, we evaluated the chronic efficacy of a neuropeptide Y(5) receptor antagonist in high-fat diet-fed neuropeptide Y(1) receptor knockout mice in order to mimic chronic combination treatment with neuropeptide Y(1) and Y(5) receptor antagonists. The neuropeptide Y(5) receptor antagonist produced greater body weight reductions in high-fat diet-fed neuropeptide Y(1) receptor knockout mice than in wild-type C57BL/6 mice. These findings confirm an interaction between neuropeptide Y(1) and Y(5) receptors in the regulation of energy homeostasis, as blockade of both the neuropeptide Y(1) and Y(5) receptors produced a greater anti-obesity effect than blocking either receptor alone.

Identification and characterization of a selective radioligand for melanin-concentrating hormone 1-receptor (MCH1R).

We have developed and characterized [(35)S]4a as a potent and selective radioligand for melanin-concentrating hormone 1-receptor (MCH1R). Compound [(35)S]4a showed appreciable specific signals in brain slices prepared from wild-type mice but not from MCH1R deficient mice, confirming the specificity and utility of [(35)S]4a as a selective MCH1R radioligand for ex vivo receptor occupancy assays.

Chronic intracerebroventricular infusion of nociceptin/orphanin FQ produces body weight gain by affecting both feeding and energy metabolism in mice.

Nociceptin/orphanin FQ (N/OFQ), an endogenous ligand for opioid receptor-like 1 (ORL1), is involved in various central functions, such as pain, psychological stress, locomotor activity, learning and memory, and feeding regulation. Of these functions, the role of N/OFQ in the regulation of feeding has been suggested by the fact that the central administration of N/OFQ leads to feeding behavior. However, the manner in which N/OFQ influences body weight control and subsequent obesity is unclear. To clarify the involvement of N/OFQ in the development of obesity, we evaluated the effects of intracerebroventricular infusion of N/OFQ on food intake and body weight in C57BL/6J mice that were fed a regular chow diet or moderately high-fat (MHF) diet (32.6% kcal fat). N/OFQ significantly increased food intake and body weight both in the regular diet- and MHF diet-fed mice, and these changes were more apparent in the MHF diet-fed mice. When we performed a pair-feeding study in N/OFQ intracerebroventricularly infused mice, N/OFQ did not cause body weight gain but increased white adipose tissue weight and plasma leptin, insulin, and cholesterol levels. N/OFQ reduced rectal temperature in pair-fed mice, in keeping with decreased UCP1 mRNA expression in brown adipose tissue. These results suggest that N/OFQ contributes to the development of obesity not only by inducing hyperphagia but also by decreasing energy expenditure.

Plasticity and injury in the developing brain.

The child's brain is more malleable or plastic than that of adults and this accounts for the ability of children to learn new skills quickly or recovery from brain injuries. Several mechanisms contribute to this ability including overproduction and deletion of neurons and synapses, and activity-dependent stabilization of synapses. The molecular mechanisms for activity-dependent synaptic plasticity are being discovered and this is leading to a better understanding of the pathogenesis of several disorders including neurofibromatosis, tuberous sclerosis, Fragile X syndrome and Rett syndrome. Many of the same pathways involved in synaptic plasticity, such as glutamate-mediated excitation, can also mediate brain injury when the brain is exposed to stress or energy failure such as hypoxia-ischemia. Recent evidence indicates that cell death pathways activated by injury differ between males and females. This new information about the molecular pathways involved in brain plasticity and injury are leading to insights that will provide better therapies for pediatric neurological disorders.

Symmetrical central tegmental tract (CTT) hyperintense lesions on magnetic resonance imaging in children.

The central tegmental tract (CTT) is mainly the extrapyramidal tract connecting between the red nucleus and the inferior olivary nucleus. There are only a few case reports describing CTT abnormalities on magnetic resonance imaging (MRI) in children. Our purpose was to evaluate the frequency of CTT lesions and their characteristics on MRI, and to correlate the MR imaging findings with clinical features. We reviewed retrospectively the MR images of 392 children (215 boys and 177 girls) ranging in age from 1 to 6 years. To evaluate symmetrical CTT hyperintense lesions, we defined a CTT lesion as an area of bilateral symmetrical hyperintensity in the tegmentum pontis on both T2-weighted images and diffusion-weighted images in more than two slices. We measured the ADC (apparent diffusion coefficient) values of symmetrical CTT hyperintensity, and compared them with those of children without CTT abnormality. CTT lesions were detected in 20 (5.1%) of the 392 children. The mean ADC value for these 20 children was significantly lower than that of the normal CTT (p<0.001). On MR imaging, other than CTT lesions, associated parenchymal lesion included: none (n=6); other abnormalities, including periventricular leukomalacia (n=3); thin corpus callosum (n=3); ventricular dilatation (n=2); encephalopathy (n=2). Clinically, cerebral palsy was the most frequent clinical diagnosis (n=6), accounting for 30%, which was significantly more frequent than the prevalence of cerebral palsy among children without CTT lesions (13%) (n<0.05). CTT lesions were detected in 5.1% of all the children examined. Cerebral palsy was the most frequent clinical diagnosis.

Effects of a novel Y5 antagonist in obese mice: combination with food restriction or sibutramine.

OBJECTIVE: To further address the function of the Y5 receptor in energy homeostasis, we investigated the effects of a novel spironolactone Y5 antagonist in diet-induced obese (DIO) mice. METHODS AND PROCEDURES: Male C57BL/6 or Npy5r(-/-) mice were adapted to high-fat (HF) diet for 6-10 months and were submitted to three experimental treatments. First, the Y5 antagonist at a dose of 10 or 30 mg/kg was administered for 1 month to DIO C57BL/6 or Npy5r(-/-) mice. Second, the Y5 antagonist at 30 mg/kg was administered for 1.5 months to DIO C57BL/6 mice, and insulin sensitivity was evaluated using an insulin tolerance test. After a recovery period, nuclear magnetic resonance measurement was performed to evaluate body composition. Third, DIO mice were treated with the Y5 antagonist alone, or in combination with 10% food restriction, or with another anorectic agent, sibutramine at 10 mg/kg, for 1.5 months. Plasma glucose, insulin, and leptin levels, and adipose tissue weights were quantified. RESULTS: The spironolactone Y5 antagonist significantly reduced body weight in C57BL DIO mice, but not in Npy5r(-/-) DIO mice. The Y5 antagonist produced a fat-selective loss of body weight, and ameliorated obesity-associated insulin resistance in DIO mice. In addition, the Y5 antagonist combined with either food restriction or sibutramine tended to produce greater body weight loss, as compared with single treatment. DISCUSSION: These findings demonstrate that the Y5 receptor is an important mediator of energy homeostasis in rodents.

A pair-feeding study reveals that a Y5 antagonist causes weight loss in diet-induced obese mice by modulating food intake and energy expenditure.

Neuropeptide Y (NPY) is thought to have a significant role in the physiological control of energy homeostasis. We recently reported that an NPY Y5 antagonist inhibits body weight gain in diet-induced obese (DIO) mice, with a moderate reduction in food intake. To clarify the mechanism of the antiobesity effects of the Y5 antagonist, we conducted a pair-feeding study in DIO mice. The Y5 antagonist at 100 mg/kg produced a moderate feeding suppression leading to an 18% decrease in body weight, without altering body temperature. In contrast, the pair-fed group showed only a transient weight reduction and a reduced body temperature, thus indicating that the Y5 antagonist stimulates thermogenesis. The Y5 antagonist-treated mice showed an up-regulation of uncoupling protein mRNA in brown adipose tissue (BAT) and white adipose tissue (WAT), suggesting that both BAT and WAT contribute to energy expenditure. Thus, the Y5 antagonist induces its antiobesity effects by acting on both energy intake and expenditure.

A neuropeptide Y Y5 antagonist selectively ameliorates body weight gain and associated parameters in diet-induced obese mice.

Neuropeptide Y (NPY) is thought to have a major role in the physiological control of energy homeostasis. Among five NPY receptors described, the NPY Y5 receptor (Y5R) is a prime candidate to mediate some of the effects of NPY on energy homeostasis, although its role in physiologically relevant rodent obesity models remains poorly defined. We examined the effect of a potent and highly selective Y5R antagonist in rodent obesity and dietary models. The Y5R antagonist selectively ameliorated diet-induced obesity (DIO) in rodents by suppressing body weight gain and adiposity while improving the DIO-associated hyperinsulinemia. The compound did not affect the body weight of lean mice fed a regular diet or genetically obese leptin receptor-deficient mice or rats, despite similarly high brain Y5R receptor occupancy. The Y5R antagonist acts in a mechanism-based manner, as the compound did not affect DIO of Y5R-deficient mice. These results indicate that Y5R is involved in the regulation and development of DIO and suggest utility for Y5R antagonists in the treatment of obesity.

Long-term effects of music therapy on elderly with moderate/severe dementia.

Over a period of 2 years we assessed the long-term effects of group music therapy carried out once weekly on the elderly (mean age: 83 years) suffering from moderate or severe dementia by observing changes in the cortisol level in saliva and in blood pressure and by an intelligence assessment. Systolic blood pressure determined 1 and 2 years after the start of therapy increased significantly in the nonmusic therapy group compared with that in music therapy group (p < .05). Systolic blood pressure increases with aging; the systolic blood pressure was significantly lower in participants who received music therapy. No significant differences in cortisol level in saliva or intelligence assessment score were observed, but the music therapy group maintained their physical and mental states during the 2-year period better than the nonmusic therapy group. This result indicates the lasting effect of once-a-week continuous music therapy. Even the elderly with moderate or severe dementia were able to participate in the group music therapy, and results suggest that enjoying singing and playing musical instruments in a concert was effective in preventing cardiac and cerebral diseases.

PARP-1 gene disruption in mice preferentially protects males from perinatal brain injury.

Poly(ADP-ribose) polymerase-1 is over-activated in the adult brain in response to ischemia and contributes to neuronal death, but its role in perinatal brain injury remains uncertain. To address this issue, 7-day-old wild-type (wt) and PARP-1 gene deficient (parp+/- and parp-/-) Sv129/CD-1 hybrid mice were subjected to unilateral hypoxia-ischemia and histologic damage was assessed 10 days later by two evaluators. Poly(ADP-ribose) polymerase-1 knockout produced moderate but significant (p < 0.05) protection in the total group of animals, but analysis by sex revealed that males were strongly protected (p < 0.05) in contrast to females in which there was no significant effect. Separate experiments demonstrated that PARP-1 was activated over 1-24 h in both females and males after the insult in neonatal wt mice and rats using immnocytochemistry and western blotting for poly(ADP-ribose). Brain levels of NAD+ were also significantly reduced, but the decrease of NAD+ during the early post-hypoxia-ischemia (HI) phase was only seen in males. The results indicate that hypoxia-ischemia activates Poly(ADP-ribose) polymerase-1 in the neonatal brain and that the sex of the animal strongly influences its role in the pathogenesis of brain injury.

[Health of aged people living with siblings who are professional nurses in China from the view point of lifestyle and health condition].

PURPOSE: Since 1978 in China, rapid economic development has taken place and the nation's quality of life has improved through the introduction of reform-opening policies. Such change has caused new health problems, partially due to aging of the population, with increase in lifestyle-related diseases and environmental pollution, and also expansion of regional variation. In this study, diseases undergoing treatment (relevance) and lifestyles of the elderly living with siblings who are professional nurses were evaluated. METHOD: We conducted a study in 23 provinces to discern characteristics and factors related to lifestyle and situations of patients undergoing treatment. We analyzed 1,548 senior citizens (response rate: 82.1%) over 65 years old living with a sibling who is a professional nurse. The professional nurse provided the replying to the questions. RESULTS: 1. A total of 457 out of 597 males (76.5%) and 725 out of 951 females (76.2%) had diseases under treatment. Males over 75 years old suffered from arteriosclerosis, cerebrovascular diseases, and heart disease. Females over 75 years old suffered from arteriosclerosis, respiratory diseases, and eye diseases. 2. In both males and females over 75 years old (older elderly) there were no significant differences in the Health Practice Index (HPI) from persons under 75 (younger elderly). Older elderly were more likely to snack often. Among males and females, 5 of 8 health-practices, such as a napping and physical exercise, differed. Females were less likely to smoke and drink alcohol. 3. In both males and females, non-diseased participants had a higher HPI than that of diseased participants. This tendency was the same in both younger and older elderly. 4. Cluster analyses of patterns of diseases revealed that the 23 provinces could be classified into 4 areas. The HPI in areas with a low proportion of diseased subjects was significantly higher than that in areas with a high proportion of diseased. One of the areas' HPI appeared to be noticeably lower than that of the other 3 and the number of participants with low physical exercise was higher in this case. CONCLUSION: Our data indicate that individuals having a high HPI appear less likely to develop lifestyle-related diseases. In 4 areas divided by the cluster analysis of patterns of diseases, there were significant differences in HPI, smoking and physical exercise.

Multiple coronary artery fistulas with a huge right coronary artery showing exacerbation during 16 years of follow-up.

A 38-year-old Japanese woman underwent cardiac evaluation to assess an arrhythmia detected after the delivery of her fourth baby. At the age of 22 years, she had undergone cardiac evaluation, including catheter examination, because of a heart murmur. Coronary angiography showed a huge right coronary artery (RCA) draining to the right atrium and a normal left coronary artery. Because the left to right shunt ratio was trivial and the patient was asymptomatic, conservative observation was undertaken. On the current admission to hospital, there was a grade III continuous murmur at the second left sternal border. The patient underwent right and left cardiac catheter examination in March 2000. Selective coronary angiography demonstrated an enlarged lumen of the RCA, an enlarged aneurysmal cavity in the terminal portion of the RCA, and the postero-descending artery being filled by collateral circulation from the left coronary artery. Moreover, 2 new fistulas had appeared from the left coronary system. An oxygen saturation study showed that the pulmonary to somatic flow ratio (Qp/Qs) was 2.2. The patient underwent surgical treatment in July 2000 and on coronary angiography 1 month later, there was no abnormal shunt flow from either coronary artery into the aneurysmal cavity, although the RCA was still enlarged.

Protective effect of erythropoietin in neonatal hypoxic ischemia in mice.

The effect of systemic erythropoietin pretreatment on hypoxic ischemic injury was examined in neonatal mice. Injury was significantly less in cortex, hippocampus, striatum and thalamus of erythropoietin-treated animals (5 U/g vs vehicle) 24 h after hypoxic ischemia and in all of these regions except hippocampus at 7 days. Activated caspase-3- and activated NFkappaB-immunoreactive neurons were observed in the injured areas; these areas were smaller in the erythropoietin group. To our knowledge, this is the first report demonstrating persistent neuroprotective effects of erythropoietin in neonatal mice.