Mutational analysis of CACNA1G in idiopathic generalized epilepsy. Mutation in brief #962. Online

Recent studies have strongly implicated low voltage-activated/T-type calcium channels (T-channels) in the etiology of epilepsy. Here, we report the results of a mutational analysis of the CACNA1G gene, encoding the T-channel Ca(V)3.1/(1G) subunit, using a cohort of 123 mostly Japanese and Hispanic patients with idiopathic generalized epilepsies (IGE) and 360 healthy control individuals. We found 13 variants, including five which involved amino acid substitutions. One variant, c.1709C>T (Ala570Val), is present in a sporadic case of juvenile myoclonic epilepsy (JME) with early childhood absence and astatic seizures, but was not found in control samples. Another variant, c.3265G>T (Ala1089Ser), was observed in three family members affected with JME, and also in one control individual. Two JME patients and three control individuals harbored a third variant, c.2968G>A (Asp980Asn). Although not statistically significant, slightly faster inactivation decay rates were observed in some mutant channels. Our collective findings flag CACNA1G as a potential susceptibility locus for IGE subsyndromes that warrants closer investigation.     

Inhibitory effect of regucalcin on Ca-dependent protein kinase activity in rat brain cytosol: involvement of endogenous regucalcin

The effect of regucalcin, a Ca²⁺-binding protein, on Ca²⁺-dependent protein kinase activity in the brain cytosol of rats with different ages (5 and 50 weeks old) was investigated. The addition of calmodulin (10 μg/ml) or dioctanoylglycerol (5 μg/ml) in the enzyme reaction mixture caused a significant increase in protein kinase activity in the presence of CaCl₂ (1 mM), indicating that Ca²⁺ calmodulin or protein kinase C is present in the cytosol. Such an increase was completely prevented by the addition of regucalcin (10⁻⁷ M). Moreover, regucalcin (10⁻⁷ M) significantly inhibited cytosolic protein kinase activity without Ca²⁺/calmodulin or dioctanoylglycerol addition. Meanwhile, the presence of anti-regucalcin monoclonal antibody (10–50 ng/ml) in the enzyme reaction mixture caused a significant elevation of protein kinase activity, suggesting an inhibitory effect of endogenous regucalcin. Brain cytosolic protein kinase activity was significantly elevated by increasing age (50-week-old rats). Also, regucalcin (10⁻⁷ M) significantly decreased protein kinase activity without Ca²⁺ addition in the brain cytosol of aged rats. However, the effect of anti-regucalcin monoclonal antibody (50 ng/ml) in elevating protein kinase activity was not seen in the brain cytosol of aged rats. These results suggest that regucalcin has an inhibitory effect on Ca²⁺-dependent protein kinase activity in rat brain cytosol, and that the effect of endogenous regucalcin may be weakened in the brain cytosol of aged rats.     

Immunoenzymatic visualization of tetrodotoxin (TTX) in Cephalothrix species (Nemertea: Anopla: Palaeonemertea: Cephalotrichidae) and Planocera reticulata (Platyhelminthes: Turbellaria: Polycladida: Planoceridae).

Tetrodotoxin (TTX) was localized as brown color in different tissues of an undescribed species of the nemertean genus Cephalothrix (phylum Nemertea) and a turbellarian Planocera reticulata (phylum Platyhelminthes) on light microscopy by means of a monoclonal anti-TTX antibody. In the Cephalothrix sp., TTX was recognized in the vesicles apically arranged in the bacillary cells in the epidermis, basal lamina, the granular cells in the proboscis epithelium, rhynchocoel epithelium, and the vesicles in the basal portion of the intestinal wall near the blood vessels and rhynchocoel. The excretory system and the ovum also showed positive reaction of TTX antigen-antibody. On the other hand, the hermaphrodite flatworm P. reticulata exhibited TTX antigen-antibody complex only in their ovum. To our knowledge, this is the first experimental effort on micro-distribution of TTX in invertebrates.     

Involvement of lipoic acid in plasma metabolites, hepatic oxygen consumption, and metabolic response to a beta-agonist in broiler chickens

The present study was conducted to determine the role of alpha-lipoic acid (LA) in plasma metabolites, hepatic O2 consumption, and beta-adrenergic response in broilers. In Expt 1, 12-d-old female broiler chicks were divided into three dietary groups and fed on diets with or without LA (5 or 50 mg/kg) until 4 or 6 weeks of age, as a 2 x 3 factorial arrangement. The dietary LA had no effect on growth rates (body weight, abdominal fat, breast muscle, and liver). The higher level of LA increased plasma non-esterified fatty acid and decreased plasma triacylglycerol concentrations only at 6 weeks of age. A 42% increase in hepatic respiration was observed in the 4-week-old chickens given 50 mg LA/kg diet. In Expt 2, 3-d-old female broiler chicks were treated with or without dietary LA at 50 mg/kg. At 30 and 31 d old, isoproterenol (2 mg/kg body weight per h) was continuously infused into a wing vein for 2 h, and changes in plasma glucose, triacylglycerol, and non-esterified fatty acid concentrations were analysed. Isoproterenol increased plasma glucose over basal levels maximally at 60 min. Furthermore, the glucose increase in the LA-treated chickens was 35% greater than that of the controls at this time. Plasma non-esterified fatty acid and triacylglycerol concentrations were decreased by the isoproterenol infusion, regardless of LA administration. Therefore, the present study suggests that dietary LA has repartitioning effects on energy metabolism in chickens (although this depends on age-related metabolic state) and is a possible facilitator in the beta-adrenergic response of plasma glucose to a beta-agonist.     

Effect of lactulose on calcium and magnesium absorption: a study using stable isotopes in adult men

To evaluate the effect of lactulose on calcium (Ca) and magnesium (Mg) absorption, we performed a clinical trial with a double-blind, randomized, crossover design in 24 healthy adult male volunteers. The absorptions of Ca and Mg were evaluated by a single-labeling method using stable isotopes. The test foods, containing lactulose at a dose of 0 g (placebo), 2 g (low-dose), or 4 g (high-dose) together with 300 mg of Ca containing 20 mg of 44Ca, and 150 mg of Mg containing 28 mg of 25Mg, were administered orally. Urine samples were collected for 8 h after the ingestion of the test food. The ratios of stable isotopes in urine (44Ca/40Ca and 25Mg/24Mg) were measured by ICP-MS (inductively coupled plasma-mass spectrometry). The urinary stable-isotopes ratios (44Ca/40Ca and 25Mg/24Mg) increased with lactulose dosage. Significant differences were observed in the Ca ratio between placebo and high-dose lactulose (p<0.01), and in the Mg ratio between placebo and low-dose lactulose and between placebo and high-dose lactulose (p<0.01). Lactulose ingestion did not change the levels of bone-resorption markers (type I collagen cross-linked N-telopeptide and deoxypyridinoline) in urine. The test foods did not cause any side effects. This study demonstrates that lactulose enhances the absorptions of Ca and Mg in adult men.     

Penetrating keratoplasty and cataract surgery

Of 46 eyes undergoing simultaneous penetrating keratoplasty and cataract extraction, 25 grafts (54%) remained clear with an average follow-up of 37 months. This low success rate can be party explained by the fact that more than 50% of our patients had unfavourable ocular conditions preoperatively. In case of simultaneous procedures, methods of cataract extraction (ICCE or ECCE) did not have any significant affect on the rate of clear grafts. In eyes with favourable ocular conditions preoperatively, the rates of clear grafts were similar (approximately 75%) in cases of simultaneous operation (21 eyes) and separate operation (8 eyes).     

Development Of The Human Mesencephalic Trigeminal Nucleus: A Morphometric Study

Anatomical development of the human mesencephalic trigeminal nucleus was assessed quantitatively and qualitatively, using serial sections of brains, with the help of a computer-assisted image-analyser. Five fetuses at 16, 18, 21, 23 and 27 weeks of gestation; five neonates at 30, 32, 33, 35 and 40 weeks of gestation; a two-month-old infant and a 63-year-old adult were studied. Morphometric analyses of the mesencephalic trigeminal nucleus revealed that orocaudal columnar length and neuronal areas gradually increased with gestational age. Diversity of neuronal areas appeared after 32 weeks of gestation, but neuronal numbers and shapes showed little change throughout prenatal and postnatal life.     

Functional neural reorganization following ischemic stroke in the precentral knob: an fMRI study]

We evaluated a 63 year-old, right-handed woman by functional MRI (fMRI) in the early and the recovery stages following a cerebral infarction in the right precentral knob. An activated signal in the ipsilateral sensorimotor cortex and contralateral supplementary motor cortex during deteriorated hand grasping (left) was observed in the early stages, whereas being unable to detect in the functionally recovered period. When simultaneous bilateral hand grasping was performed, the activated signal around infarct region was enlarged in recovered period. The functionally neural reorganization processes relating recovered hand movement after localized cortical infarction (precentral knob) was suggested.