Anticonvulsant profile and teratogenic evaluation of potent new analogues of a valproic acid urea derivative in NMRI mice

BACKGROUND: Valproic acid (VPA) is used to treat epilepsy and bipolar disorders, as well as for migraine prophylaxis. However, its clinical use is limited by two life-threatening side effects: hepatotoxicity and teratogenicity. To develop a more potent and safer second-generation VPA drug, the urea derivatives of four VPA analogs (2-ethyl-3-methylpentanoyl urea, 2-ethylhexanoyl urea, 2-ethyl-4-methylpentanoyl urea, and 2-methylbutanoyl urea) were synthesized. METHODS: Four CNS-active analogs of a VPA urea derivative testedthe anticonvulsant activity in the maximal electroshock seizure test (MES) and subcutaneous metrazol seizure threshold test (scMet). Teratogenic effects of these compounds were evaluated in NMRI mice susceptible to VPA-induced teratogenicity by comparison with VPA. RESULTS: All four VPA analogs showed superior anticonvulsant activity over VPA. Compared with VPA, which induced neural tube defects (NTDs) in fetuses at 1.8 and 3.6 mmol/kg, the analog derivatives induced no NTDs at any concentration up to 4.8 mmol/kg (except for a single abnormality at 3.6 mmol/kg with 2-ethyl-3-methylpentanoyl urea). Skeletal examination also revealed that the acylurea derivatives induced vertebral and rib abnormalities in fetuses markedly less frequently than VPA. Our results confirmed that the analogue derivatives are significantly less teratogenic than VPA in NMRI mice. CONCLUSIONS: The CNS-active VPA analogs containing a urea moiety, which have better anticonvulsant potency and lack teratogenicity, are good potential candidates as second-generation VPA antiepileptic drugs. Birth Defects Res (Part B) 86:394–401, 2009. © 2009 Wiley-Liss, Inc.     

Spatial Prioritization for Biodiversity Restoration: A Simple Framework Referencing Past Species Distributions

The spatial prioritization of biodiversity restoration sites on a national scale lags behind that of protected areas. We present a simple framework for spatial restoration prioritization using complementarity analysis based on the differences between past and present species distributions and restoration feasibility. In the framework, sites that were not only diverse or unique in the past (high restoration potential), but also have high restoration feasibility (e.g. low anthropogenic pressures) are ranked higher. We applied the framework to endangered bird species that breed in Japan for which distribution data were available from nationwide distribution censuses conducted in 1978 and 1998. The data were compiled in a 20‐km grid resolution across Japan. We incorporated the degree of urbanization and number of farmers as a cost to optimize the sets of cells with regard to restoration feasibility. We successfully identified candidate sets of restoration cells, in which at most six can be allocated for each species under the constraint that the total area selected does not exceed 15% of the degraded cells. At most, 11 cells were allocated for each species under the constraint that restoration cells were within 15% of all ecosystem types. Comparison with the distribution of existing nature restoration sites revealed that only 9.3 or 9.7% (depended on the constraint) of high‐priority cells overlapped cells including existing restoration sites. Our framework provides an objective, efficient method to obtain a testable plan for biodiversity restoration, which might contribute to the attainment of the 2020 target set by the Convention on Biological Diversity .     

Decreased thermodynamic stability as a crucial factor for familial amyloidotic polyneuropathy

A single mutation in the wild-type transthyretin (WT TTR) such as V30M causes a familial amyloidotic polyneuropathy disease. Comparison of the three-dimensional crystal structures of WT and V30M does not tell much about the reason. High-pressure NMR revealed that at neutral pH both WT and V30M exist as equilibrium between the native tetramer and the dissociated/unfolded monomer. The native tetramer is highly stable in WT (deltaG(0)=104 kJ/mol at 37 degrees C, pH 7.1), but the stability is significantly reduced in V30M (deltadeltaG(0)=-18 kJ/mol), increasing the fraction of the unfolded monomer by a 1000-fold. Significant reduction of thermodynamic stability of WT TTR by mutation could be a crucial factor for familial amyloidotic polyneuropathy.     

Influence of rainfall on sleeping site choice by a group of anubis baboons (Papio anubis)

For diurnal nonhuman primates, shifting among different sleeping sites may provide multiple benefits such as better protection from predators, reduced risk of parasitic infection, and closer proximity to spatially and temporally heterogeneous food and water. This last benefit may be particularly important in sleeping site selection by primates living in savanna‐woodlands where rainfall is more limited and more seasonally pronounced than in rainforests. Here, we examined the influence of rainfall, a factor that affects food and water availability, on the use of sleeping sites by anubis baboons (Papio anubis) over two 13‐month study periods that differed in rainfall patterns. We predicted that during wet periods, when food and water availability should be higher, the study group would limit the number of sleeping sites and would stay at each one for more consecutive nights than during dry periods. Conversely, we predicted that during dry periods the group would increase the number of sleeping sites and stay at each one for fewer consecutive nights as they searched more widely for food and water. We also predicted that the group would more often choose sleeping sites closer to the center of the area used during daytime (between 07:00 and 19:00) during wet months than during dry months. Using Global Positioning System data from collared individuals, we found that our first prediction was not supported on either monthly or yearly timescales, although past monthly rainfall predicted the use of the main sleeping site in the second study period. Our second prediction was supported only on a yearly timescale. This study suggests that baboons’ choice of sleeping sites is fluid over time while being sensitive to local environmental conditions, one of which may be rainfall.     

Qualitative variation in roadside weed vegetation along an urban–rural road gradient

Information on mechanisms of differentiation or homogenization of urban floras is deficient, despite their importance for urban nature conservation and urban land management. Roads, as a major human promoter of urbanization, can be an initial habitat for plants dispersed by transportations. We assumed that variation in weed vegetation along urban–rural roadside gradients is small, particularly in curbside cracks. We classified vegetation occurring in curbside cracks along the National Route 3 in southern Japan and compared the characteristics of the vegetation types recognized. Species in curbside cracks were recorded on 40 plots. Three vegetation types were classified, in part related to surrounding land-use types. Although the Shannon–Wiener diversity index and the number of native species clearly differed among the vegetation types in the curbside cracks, no significant differences between the three floristic vegetation types were found in the number of non-natives and invasive alien species. This may result from the small specificity and complexity of landscape structures, due to the continuity and connectivity of paved-road networks. Of the 122 species, 44 were non-natives or invasive alien species. The vegetation types generally involved the same plant families, with large numbers of species from the Asteraceae and Poaceae, regardless of vegetation types, but frequency of occurrence of the two families clearly differed between natives and non-natives and invasive alien species. Ephemeral non-natives and invasive alien species, particularly Asteraceae and Poaceae, seem to have advantageous dispersal strategies or low habitat dependency facilitating their occurrence in curbside cracks, regardless of adjacent land uses and the urban–rural landscape gradient. Expansion of these species may cause a homogenization of regional floras along roads.     

Slow conformational dynamics in the hamster prion protein

Although the mechanism of the conformational conversion from the cellular (PrP(C)) to the scrapie (PrP(Sc)) form of animal prion proteins has yet to be elucidated, evidence is accumulating that may provide insight into the conversion process at atomic resolution. Here we show critical aspects of the slow fluctuation dynamics of the recombinant hamster prion protein, rPrP(90-231), based on NMR relaxation analysis using Carr-Purcell-Meiboom-Gill (CPMG) experiments, and compare them in detail with results from high-pressure NMR. Residues exhibiting slow fluctuations on the time scale of microseconds to milliseconds are mainly localized on helices B and C (172-193 and 200-227), which include locally disordered regions in an intermediate conformer, PrP*, identified previously by high-pressure NMR [Kuwata, K., et al., (2002) Biochemistry 41, 12277-12283]. Moreover, chemical shift differences between two putative exchanging conformers obtained by the CPMG relaxation analysis and the linear component of the pressure-induced chemical shift changes are reasonably correlated at individual residue sites. These observations suggest that both the CMPG relaxation and the pressure shifts reflect slow conformational fluctuations and that these slow motions in PrP(C) are related to the trajectories leading to the transition to PrP*.     

Comparisons of recruitment, survival, and growth in invasive and native saplings on a volcano

We monitored the recruitment, survival, and growth of tree saplings on invasive (Larix kaempferi) versus native species (Betula and Populus) using 16 20 m × 20 m plots established along elevation gradient on the volcano Mount Koma, Japan, for 7 years because the sapling behaviors should determine forest structures. The crowding of overstory consists mostly of Larix decreased with increasing elevation. Larix recruits were conspicuous, particularly at middle elevation where overstory crowding was intermediate, while Betula recruits were least. Larix overstory crowding inhibited the recruitment of all the taxa, although intermediate crowding promoted the recruitment of Larix. The restriction of sapling emergence was conspicuous at lower elevation where the overstory crowding was highest, probably because of shading, and/or competition with overstory trees. Sapling recruitment for all taxa was restricted at higher elevation, due to high stresses derived from direct solar radiation and strong wind without overstory. The survival of saplings was 96% for Larix and Betula, while it was ca. 50% for Populus. Larix overstory decreased the survival and growth of all the taxa, except Larix survival and Betula growth. The results implied that Larix could establish by high survival once the recruits succeeded everywhere and native sapling regeneration was restricted by Larix overstory. Strong recruitment, survival, and growth of Larix, together with resistance to overstory crowding, enables it to dominate and persist in such disturbed areas regardless of the canopy closure.     

Organ-specific and age-dependent expression of insulin-like growth factor-I (IGF-I) mRNA variants: IGF-IA and IB mRNAs in the mouse

Insulin-like growth factor-I (IGF-I) gene generates several IGF-I mRNA variants by alternative splicing. Two promoters are present in mouse IGF-I gene. Each promoter encodes two IGF-I mRNA variants (IGF-IA and IGF-IB mRNAs). Variants differ by the presence (IGF-IB) or absence (IGF-IA) of a 52-bp insert in the E domain-coding region. Functional differences among IGF-I mRNAs, and regulatory mechanisms for alternative splicing of IGF-I mRNA are not yet known. We analyzed the expression of mouse IGF-IA and IGF-IB mRNAs using SYBR Green real-time RT-PCR. In the liver, IGF-I mRNA expression increased from 10 days of age to 45 days. In the uterus and ovary, IGF-I mRNA expression increased from 21 days of age, and then decreased at 45 days. In the kidney, IGF-I mRNA expression decreased from 10 days of age. IGF-IA mRNA levels were higher than IGF-IB mRNA levels in all organs examined. Estradiol-17beta (E2) treatment in ovariectomized mice increased uterine IGF-IA and IGF-IB mRNA levels from 3 hr after injection, and highest levels for both mRNAs were detected at 6 hr, and relative increase was greater for IGF-IB mRNA than for IGF-IA mRNA. These results suggest that expression of IGF-I mRNA variants is regulated in organ-specific and age-dependent manners, and estrogen is involved in the change of IGF-I mRNA variant expression.