High-frequency electrically stimulated skeletal muscle contractions increase p70 phosphorylation independent of known IGF-I sensitive signaling pathways

Insulin-like growth factor (IGF-I) is hypothesized to be a critical upstream regulator of mammalian target of rapamycin (mTOR)-regulated protein synthesis with muscle contraction. We utilized a mouse model that expresses a skeletal muscle specific dominant-negative IGF-I receptor to investigate the role of IGF-I signaling of protein synthesis in response to unilateral lengthening contractions (10 sets, 6 repetitions, 100 Hz) at 0 and 3 h following the stimulus. Our results indicate that one session of high frequency muscle contractions can activate mTOR signaling independent of signaling components directly downstream of the receptor.     

Food resource allocation patterns in lactating females in a long-term selection experiment for litter size in mice

Resource allocation patterns, as quantified by residual food intake (RFI), and the consequences for offspring development were investigated during lactation in 96 females of a mouse line selected for 104 generations for high litter size at birth (S-line) and in 87 females of a non-selected control line (C-line). Litters of 45 C-line dams (Cs) and 48 S-line dams (Ss) were standardised (s) at birth; other dams (ns) supported total number of pups born (Cns and Sns, respectively). RFI during lactation was significantly lower in Sns-dams than in C-line dams and Sns-dams. After weaning Sns-dams seemed to be able to restore the negative resource situation. Sns-pups were about 25% less mature than Cns-pups at all times. Maturity was similar for Cs- and Ss-pups from 2 d in lactation on, and about 18% and 53% higher than Cns- and Sns-pups. The pre-weaning mortality rate was significantly higher in Sns-litters (35.6 ± 2.76) than in Cns-litters (4.95 ± 2.23). The results suggest that S-line dams allocated considerably more resources to maintenance of offspring than C-line dams. This was insufficient to provide the offspring with an adequate amount of resources, resulting in reduced pup development and increased pre-weaning mortality rates.     

Synthesis and biological activity of certain 1,2,3-triazole carboxamide nucleosides related to bredinin and pyrazofurin

5-Hydroxy-1-(beta-D-ribofuranosyl)-1,2,3-triazole-4-carboxamide (II) has been prepared by direct glycosylation of the trimethylsilyl derivative of 5-hydroxy-1,2,3-triazole-4-carboxamide (IV). The use of trimethylsilyltrifluoromethane sulfonate as a catalyst and 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose in acetonitrile gave a 95% yield of a 1:1 mixture of V and VI as blocked nucleosides which were readily separated on a silica gel column. The synthesis of II was also achieved by the cycloaddition of 2,3,5-tri-O-benzoyl-beta-D-ribofuranosyl azide with ethyl malonamate. II shows significant antiviral activity against herpes and measles virus in vitro.     

硫氧还蛋白抗体柱的制备

目的:探索硫氧还蛋白(Trx)抗体柱对Trx融合蛋白纯化的可行性。方法与结果:对含有Trx基因的质粒表达载体pTrxFus进行改造,在Trx读框之后加入6×His序列,并在大肠杆菌中表达C端带有6×His标签的Trx,经Ni2+柱亲和纯化后制备多克隆抗体;把经蛋白A纯化后的抗体偶联在溴化氰活化的琼脂糖凝胶上,制成Trx抗体柱;用此抗体柱纯化与Trx融合表达的豇豆胰蛋白酶抑制剂(CpTI),SDS-PAGE结果显示获得了纯度较高的Trx-CpTI。结论:用Trx抗体制成的免疫亲和层析柱可以有效纯化Trx融合蛋白。     

Are environmental factors important facilitators of pompom weed (<Emphasis Type="Italic">Campuloclinium macrocephalum</Emphasis>) invasion in South African rangelands?

The absence of natural enemies being keystone to the success of invasive alien plants (IAPs) can only be accepted once all the factors governing the invader and the ecosystems it invades have been established. Few studies have attempted this approach. This study reports on the relations between the invasive alien forb Campuloclinium macrocephalum (Less.) DC. (pompom weed), herbivory and the ecology of the rangelands invaded in South Africa. Eighty invaded rangelands in Gauteng Province were studied. Pompom weed herbivory was insignificant with the greatest damage being caused by native grasshoppers. Rainfall, topography and soil texture were the primary drivers of vegetation pattern. Pompom weed exhibited disturbance-mediated invasion strategies, favouring rangelands affected by non-sustainable commercial grazing practices, abandoned agricultural fields and drained wetlands. It invaded vegetation in poor condition with sward basal cover <19%. Weed density was exacerbated by high fire frequency. Vegetation susceptible to pompom weed was also invaded by other alien and native pasture weeds. The absence of natural enemies could give pompom weed a competitive advantage over native pioneer cohorts in disturbed rangelands, but is unlikely to increase fitness enabling its establishment in vegetation in good condition. Grazing strategies that promote the dominance of grass species adapted to frequent non-selective defoliation appear to restrict pompom weed better than degraded rangelands that are not utilised. Biotic resistance of rangelands to pompom weed invasion will be greatest in vegetation in good condition and under good management. The low abundance of pompom weed in rangelands in communal areas and fence-line contrasts in invaded and un-invaded road reserves warrants further investigation.     

In Silico Screening on the Three-dimensional Model of the Plasmodium vivax SUB1 Protease Leads to the Validation of a Novel Anti-parasite Compound

Widespread drug resistance calls for the urgent development of new antimalarials that target novel steps in the life cycle of Plasmodium falciparum and Plasmodium vivax. The essential subtilisin-like serine protease SUB1 of Plasmodium merozoites plays a dual role in egress from and invasion into host erythrocytes. It belongs to a new generation of attractive drug targets against which specific potent inhibitors are actively searched. We characterize here the P. vivax SUB1 enzyme and show that it displays a typical auto-processing pattern and apical localization in P. vivax merozoites. To search for small PvSUB1 inhibitors, we took advantage of the similarity of SUB1 with bacterial subtilisins and generated P. vivax SUB1 three-dimensional models. The structure-based virtual screening of a large commercial chemical compounds library identified 306 virtual best hits, of which 37 were experimentally confirmed inhibitors and 5 had K_i values of <50 μm for PvSUB1. Interestingly, they belong to different chemical families. The most promising competitive inhibitor of PvSUB1 (compound 2) was equally active on PfSUB1 and displayed anti-P. falciparum and Plasmodium berghei activity in vitro and in vivo, respectively. Compound 2 inhibited the endogenous PfSUB1 as illustrated by the inhibited maturation of its natural substrate PfSERA5 and inhibited parasite egress and subsequent erythrocyte invasion. These data indicate that the strategy of in silico screening of three-dimensional models to select for virtual inhibitors combined with stringent biological validation successfully identified several inhibitors of the PvSUB1 enzyme. The most promising hit proved to be a potent cross-inhibitor of PlasmodiumSUB1, laying the groundwork for the development of a globally active small compound antimalarial.     

Unusual assemblages of Late Cretaceous silicoflagellates from the Canadian Archipelago

Devon and Ellef Ringnes islands in the Canadian Arctic Archipelago contain well-preserved Santonian through early Campanian silicoflagellates. These occur in three unusual assemblages, which comprise three biostratigraphic zones. The oldest, the Vallacerta tumidula Partial Range Zone of Santonian age, contains an extremely variable microflora that includes species of the poorly-known genus Variramus, two new species of Cornua, and a new genus Umpiocha. The Schulzyocha ruppelii Range Zone of early Campanian age includes several species of the new genus Schulzyocha, reflecting quadrate silicoflagellate skeletal morphologies that lack a basal structure. The youngest assemblage includes the poorly-known species Cornua trifurcata, the nominative taxon of the C. trifurcata Partial Range Zone (proposed by McCartney et al., in press c; emended herein), which allows a correlation with a silicoflagellate-bearing sequence from the Horton River area, Northwest Territories of Canada. This report presents information on 24 silicoflagellate taxa, proposes 2 new genera, Schulzyocha and Umpiocha, and describes 13 new species and 1 new combination. All of these new taxa disappear by the top of the Campanian Cornua trifurcata Partial Range Zone. These assemblages reveal a new history of silicoflagellate evolution prior to the late Campanian, the period from which most of our prior knowledge about Cretaceous silicoflagellates was derived. The assemblages show progressive change from skeletons with highly variable morphologies to more stable silicoflagellate skeletal geometry. Two important Corbisema species, C. apiculata and C. archangelskiana, appear to be derived from separate lines within Cornua in the latest Santonian and early Campanian. The important Cretaceous genus Lyramula makes its first appearance at the beginning of the Campanian.     

Ex vivo measurement of calpain activation in human peripheral blood lymphocytes by detection of immunoreactive products of calpastatin degradation

Limited proteolysis of multiple intracellular proteins by endogenous Ca-dependent cysteine proteases--calpains--is an important regulatory mechanism for cell proliferation, apoptosis etc. Its importance for cellular functions is stressed by existence of endogenous calpain inhibitors--calpastatins. The calpain-calpastatin system within living cells is in a fragile balance, which depends on both partners. The interdependence of calpain--a protease--and calpastatin--an endogenous inhibitor and at the same time a substrate for this enzyme makes any assessment of actual activity of this enzyme in the cells very difficult. In this work we made an attempt to estimate and compare the activity of calpain in human peripheral blood lymphocytes by assessing the levels of limited proteolysis of calpastatin in these cells by western blot, while at the same time the levels of calpain protein inside these cells was measured by flow cytometry. Our results indicate that it is possible to compare (semi-quantitatively) the activities of calpain in peripheral blood CD4+ and CD19+ lymphocytes from various donors that way. Preliminary results showed that calpain activity is increased in the CD4+ T cells isolated from peripheral blood of rheumatoid arthritis patients as compared to control lymphocytes. Extremely high intrinsic activity of calpain was detected in chronic lymphocytic leukemia (CD19+) cells. All this confirms the detection of immunoreactive products of calpastatin as a good maker of endogenous calpain activity.