The voltage-gated Kv2.1 potassium channel encoded by KCNB1 produces the major delayed rectifier potassium current in pyramidal neurons. Recently, de novo heterozygous missense KCNB1 mutations have been identified in three patients with epileptic encephalopathy and a patient with neurodevelopmental disorder. However, the frequency of KCNB1 mutations in infantile epileptic patients and their effects on neuronal activity are yet unknown. We searched whole exome sequencing data of a total of 437 patients with infantile epilepsy, and found novel de novo heterozygous missense KCNB1 mutations in two patients showing psychomotor developmental delay and severe infantile generalized seizures with high-amplitude spike-and-wave electroencephalogram discharges. The mutation located in the channel voltage sensor (p.R306C) disrupted sensitivity and cooperativity of the sensor, while the mutation in the channel pore domain (p.G401R) selectively abolished endogenous Kv2 currents in transfected pyramidal neurons, indicating a dominant-negative effect. Both mutants inhibited repetitive neuronal firing through preventing production of deep interspike voltages. Thus KCNB1 mutations can be a rare genetic cause of infantile epilepsy, and insufficient firing of pyramidal neurons would disturb both development and stability of neuronal circuits, leading to the disease phenotypes.
Brown rot basidiomycetes have long been thought to lack the processive cellulases that release soluble sugars from crystalline cellulose. On the other hand, these fungi remove all of the cellulose, both crystalline and amorphous, from wood when they degrade it. To resolve this discrepancy, we grew Gloeophyllum trabeum on microcrystalline cellulose (Avicel) and purified the major glycosylhydrolases it produced. The most abundant extracellular enzymes in these cultures were a 42-kDa endoglucanase (Cel5A), a 39-kDa xylanase (Xyn10A), and a 28-kDa endoglucanase (Cel12A). Cel5A had significant Avicelase activity-4.5 nmol glucose equivalents released/min/mg protein. It is a processive endoglucanase, because it hydrolyzed Avicel to cellobiose as the major product while introducing only a small proportion of reducing sugars into the remaining, insoluble substrate. Therefore, since G. trabeum is already known to produce a -glucosidase, it is now clear that this brown rot fungus produces enzymes capable of yielding assimilable glucose from crystalline cellulose.
Production of melon (Cucumis melo) may be limited by excesses of boron and salinity, and it was hypothesized that melon grafted onto Cucurbita rootstock would be more tolerant to excessive boron concentrations than non-grafted plants. The objectives of this study were: (i) to determine the effects of salinity and excessive boron concentrations in irrigation water on growth and yields of grafted and non-grafted melon plants; and (ii) to study the interaction between the effects of salinity and boron on the uptake of macroelements and boron by grafted and non-grafted melon plants. The plants were grown in pots of Perlite in a greenhouse. The combined effects of boron and salinity on growth and yield were investigated for five boron concentrations, ranging from 0.2 to 10 mg L −1 , and two salinity levels, electrical conductivity (EC) 1.8 and 4.6 dS m −1 , in the irrigation water. With low salinity the boron concentrations in old leaves of non-grafted and grafted plants ranged from 249 to 2827 and from 171 to 1651 mg kg −1 dry weight, respectively; with high salinity the corresponding concentrations ranged from 192 to 2221 and from 200 to 1263 mg kg −1 dry weight, respectively. These results indicate that the grafted plants accumulated less boron than the non-grafted plants when they were exposed to similar boron concentrations, and that both plant types absorbed less boron when irrigated with the more saline irrigation water. It is suggested that: (i) the Cucurbita rootstock excluded some boron and that this, in turn, decreased the boron concentration in the grafted plants; and (ii) the low boron uptake under high-salinity irrigation was mainly a result of reduced transpiration of the plants. Significant negative linear regressions were found between fruit yield and leaf boron concentration for grafted plants, under both low and high salinity levels, and for non-grafted plants under low salinity. The fruit yield of the grafted plants was less affected by boron accumulation in the leaves than that of non-grafted plants. Increasing the water salinity decreased the sensitivity of both plant types to increases in leaf boron concentration, which indicates that the effects of boron and salinity on melon plants were not additive.
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