单链胰岛素前体在酵母中的分泌表达及其转变成人胰岛素

将化学合成的单链猪胰岛素前体(PIP)基因和用聚合酶链反应得到的α交配因子前导顺序(αMFL)基因插入质粒pVT102-U的醇脱氢酶基因ADH1的启动子和3’终止顺序之间而生成质粒pVT102-U/αMFL-PIP.被pVT102-U/αMFL-PIP转化的酵母(Saccharomyces cerevistae)可表达单链前体并分泌到培养基中.前体在纯化后可通过胰蛋白酶的转肽作用转变成人胰岛素.纯化的人胰岛素具有全部活力并可结晶.人胰岛素的总收率为每升培养液25mg.     

Molecular phylogeny of the springhare, Pedetes capensis, based on mitochondrial DNA sequences

The phylogenetic position of the Pedetidae, represented by a single species Pedetes capensis, is controversial, reflecting in part the retention of both Hystricomorphous and Sciurognathous characteristics in this rodent. In an attempt to clarify the species evolutionary relationships, mtDNA gene sequences from 10 rodent species (representing seven families) were analyzed using phenetic, parsimony, and maximum-likelihood methods of phylogenetic inference; the rabbit, Oryctolagus cuniculus (Order Lagomorpha), and cow, Bos taurus (Order Artiodactyla), were used as outgroups. Investigation of 714 base pairs of the protein-coding cytochrome b gene indicate strong base bias at the third codon position with significant rate heterogeneity evident between the three structural domains of this gene. Similar analyses conducted on 816 base pairs of the 12S rRNA gene revealed a transversion bias in the loop sections of all taxa. The cytochrome b gene sequences proved useful in resolving associations between closely related species but failed to produce consistent tree topologies at the family level. In contrast, phylogenetic analysis of the 12S rRNA gene resulted in strong support for the clustering of Pedetidae/Heteromyidae/Geomyidae and Muridae in one clade to the exclusion of the Hystricidae/Thryonomyidae and Sciuridae, a finding which is concordant with studies of rodent fetal membranes as well as reproductive and other anatomical features.      

Nomenclature: or what's in a name?

Transgenic Research -     

Optimization of embryo transfer protocols for mice

The efficiency of ova transfer and subsequent survivability were explored in this study. The goals of the experiment were to 1) determine the minimum number of ova necessary for pregnancy maintenance, 2) ascertain if the number of zygotes used in ova transfer approaches or exceeds uterine capacity, and 3) establish if location of deposition of ova influences embryo survival. A total of 1647 pronuclear zygotes were transferred in groups of 1, 2, 4, 6, 15 or 25 on Day 1 of gestation either via the oviducal ampulla or ostium to 156 nulliparous ICR pseudopregnant female mice. Pregnancy status was determined on Day 12 or Day 19 of gestation. Results indicated that pregnancy rates were not significantly increased by transferring larger numbers of zygotes (P < 0.1504) and that beyond transfer of 15 zygotes, the progressive increase in fetal numbers per litter declined. However, on Day 19 of gestation, no definitive evidence of limitation of uterine capacity was obtained with the numbers of zygotes transferred (P < 0.0531), and the estimates of numbers of viable and resorbed fetuses differed when determinations were made on Day 12 versus Day 19 of gestation. Mean numbers of developed fetuses per recipient declined (P < 0.0001), whereas the number of resorptions (partially resorbed fetuses or resorption sites) increased (P < 0.0001) over this period, reflecting fetal loss in mid- to late-gestation and possibly the transient nature of resorptions prior to Day 12. Additionally, there was no difference in pregnancy outcome when transferring ova into the oviducal ostium or isthmus (P < 0.5256). Finally, these results illustrated that when large numbers of zygotes were transferred into the oviducal ampulla, equivalent numbers of ova eventually implanted in the uterus; however, proportionally more of them began resorption.     

Serum concentrations of prolactin, oestrogen and LH during the perioestrous period in prepubertal gilts induced to ovulate and mature gilts

The temporal relationships of serum prolactin, oestrogen and LH concentrations during the perioestrous period were compared in prepubertal gilts induced to ovulate by PMSG and hCG and in mature gilts. In Exp. 1, 2 sustained prolactin surges, beginning 4 days and 1 day before the preovulatory LH surge, occurred in all mature gilts. A single preovulatory prolactin surge occurred in 3 prepubertal gilts, starting just before the preovulatory LH surge, but 4 prepubertal gilts had neither a prolactin nor an LH surge. A status (prepubertal or mature) versus time interaction (P less than 0.01) was detected for serum prolactin concentrations. A preovulatory oestrogen surge occurred in all gilts but was of lesser magnitude (P less than 0.01) and duration (P less than 0.05) in the prepubertal gilts without prolactin and LH surges compared to mature gilts and of lesser magnitude (P less than 0.01) compared to prepubertal gilts with prolactin and LH surges. The relative timing of the oestrogen surge in prepubertal gilts corresponded with that of mature gilts when adjusted to the LH surge (if present) but was delayed (P less than 0.01) in all prepubertal gilts if standardized to the hCG injection. In Exp. 2, mature gilts were examined to determine whether 2 perioestrous prolactin surges were characteristic of all cycling gilts. Of 9 gilts, 8 exhibited an initial prolactin surge 4-5 days before oestrus and 5/9 gilts exhibited a periovulatory prolactin surge. The presence of 2 perioestrous serum prolactin surges was not a requirement for subsequent pregnancy maintenance.(ABSTRACT TRUNCATED AT 250 WORDS)     

玉米叶片生长部位渗透调节和生长的生物物理参数变化

玉米叶片生长部位随着水分胁迫加剧ψ_w降低、LER减慢。LER从最大到零,快速干旱处理的ψw从-0.55降至-0.85 MPa;缓慢干旱处理ψ_w从-0.88降至-1.13 MPa。在任何一种LER下,缓慢干旱处理的ψ_s比快速干旱处理更低,生长停止时,前者为-1.57 MPa,而后者为-1.30MPa。缓慢干旱叶片尽管在更低ψ_w下,仍能维持一定膨压,保持一定的生长速率。经历长时间水分胁迫会改变细胞延伸生长的生物物理参数,增大临界膨压(0.08～0.09 MPa)。这是水分胁迫植株,在一定ψ_p下生长速率减慢的原因。     

Hierarchical Carbide‐Derived Carbon Foams with Advanced Mesostructure as a Versatile Electrochemical Energy‐Storage Material

Highly porous carbide‐derived carbon (CDC) mesofoams (DUT‐70) are prepared by nanocasting of mesocellular silica foams with a polycarbosilane precursor. Ceramic conversion followed by silica removal and high‐temperature chlorine treatment yields CDCs with a hierarchical micro‐mesopore arrangement. This new type of polymer‐based CDC is characterized by specific surface areas as high as 2700 m² g^?1, coupled with ultrahigh micro‐ and mesopore volumes up to 2.6 cm³ g^?1. The relationship between synthesis conditions and the properties of the resulting carbon materials is described in detail, allowing precise control of the properties of DUT‐70. Since the hierarchical pore system ensures both efficient mass transfer and high capacities, the novel CDC shows outstanding performance as an electrode material in electrochemical double‐layer capacitors (EDLCs) with specific capacities above 240 F g^?1 when measured in a symmetrical two‐electrode configuration. Remarkable capacities of 175 F g^?1 can be retained even at high current densities of 20 A g^?1 as a result of the enhanced ion‐transport pathways provided by the cellular mesostructure. Moreover, DUT‐70 can be infiltrated with sulfur and host the active material in lithium–sulfur battery cathodes. Reversible capacities of 790 mAh g^?1 are achieved at a current rate of C/10 after 100 cycles, which renders DUT‐70 an ideal support material for electrochemical energy‐storage applications.     

Tomato yellow leaf curl viruses: ménage à trois between the virus complex,the plant and the whitefly vector

Tomato yellow leaf curl disease (TYLCD) is one of the most devastating viral diseases affecting tomato crops in tropical, subtropical and temperate regions of the world. Here, we focus on the interactions through recombination between the different begomovirus species causing TYLCD, provide an overview of the interactions with the cellular genes involved in viral replication, and highlight recent progress on the relationships between these viruses and their vector, the whitefly Bemisia tabaci. Taxonomy: The tomato yellow leaf curl virus‐like viruses (TYLCVs) are a complex of begomoviruses (family Geminiviridae, genus Begomovirus) including 10 accepted species: Tomato yellow leaf curl Axarquia virus (TYLCAxV), Tomato yellow leaf curl China virus (TYLCCNV), Tomato yellow leaf curl Guangdong virus (TYLCGuV), Tomato yellow leaf curl Indonesia virus (TYLCIDV), Tomato yellow leaf curl Kanchanaburi virus (TYLVKaV), Tomato yellow leaf curl Malaga virus (TYLCMalV), Tomato yellow leaf curl Mali virus (TYLCMLV), Tomato yellow leaf curl Sardinia virus (TYLCSV), Tomato yellow leaf curl Thailand virus (TYLCTHV), Tomato yellow leaf curl Vietnam virus (TYLCVNV) and Tomato yellow leaf curl virus(TYLCV). We follow the species demarcation criteria of the International Committee on Taxonomy of Viruses (ICTV), the most important of which is an 89% nucleotide identity threshold between full‐length DNA‐A component nucleotide sequences for begomovirus species. Strains of a species are defined by a 93% nucleotide identity threshold. Host range: The primary host of TYLCVs is tomato (Solanum lycopersicum), but they can also naturally infect other crops [common bean (Phaseolus vulgaris), sweet pepper (Capsicum annuum), chilli pepper (C. chinense) and tobacco (Nicotiana tabacum)], a number of ornamentals [petunia (Petunia×hybrida) and lisianthus (Eustoma grandiflora)], as well as common weeds (Solanum nigrum and Datura stramonium). TYLCVs also infect the experimental host Nicotiana benthamiana. Disease symptoms: Infected tomato plants are stunted or dwarfed, with leaflets rolled upwards and inwards; young leaves are slightly chlorotic; in recently infected plants, fruits might not be produced or, if produced, are small and unmarketable. In common bean, some TYLCVs produce the bean leaf crumple disease, with thickening, epinasty, crumpling, blade reduction and upward curling of leaves, as well as abnormal shoot proliferation and internode reduction; the very small leaves result in a bushy appearance.     

Myofibrillar protein synthesis in myostatin-deficient mice

Either increased protein synthesis or prolonged protein half-life is necessary to support the excessive muscle growth and maintenance of enlarged muscles in myostatin-deficient mice. This issue was addressed by determining in vivo rates of myofibrillar protein synthesis in mice with constitutive myostatin deficiency (Mstn(DeltaE3/DeltaE3)) or normal myostatin expression (Mstn(+/+)) by measuring tracer incorporation after a systemic flooding dose of l-[ring-(2)H(5)]phenylalanine. At 5-6 wk of age, Mstn(DeltaE3/DeltaE3) mice had increased muscle mass (40%), fractional rates of myofibrillar synthesis (14%), and protein synthesis per whole muscle (60%) relative to Mstn(+/+) mice. With maturation, fractional rates of synthesis declined >50% in parallel with decreased DNA and RNA [total, 28S rRNA, and poly(A) RNA] concentrations in muscle. At 6 mo of age, Mstn(DeltaE3/DeltaE3) mice had even greater increases in muscle mass (90%) and myofibrillar synthesis per muscle (85%) relative to Mstn(+/+) mice, but the fractional rate of synthesis was normal. Estimated myofibrillar protein half-life was not affected by myostatin deficiency. Muscle DNA concentrations were reduced in both young and mature Mstn(DeltaE3/DeltaE3) mice, whereas RNA concentrations were normal, so the ratio of RNA to DNA was approximately 30% greater than normal in Mstn(DeltaE3/DeltaE3) mice. Thus the increased protein synthesis and RNA content per muscle in myostatin-deficient mice cannot be explained entirely by an increased number of myonuclei.     

The co-distribution of Plasmodium falciparum and hookworm among African schoolchildren

Background

On the island of Bioko (Equatorial Guinea), insecticide-treated nets (ITNs) have been the main tool used to control malaria over the last 13 years. In 2004, started an indoor residual spraying (IRS) campaign to control malaria. The purpose of this study is to asses the impact of the two control strategies on the island of Bioko (Equatorial Guinea), with regards to Plasmodium infection and anaemia in the children under five years of age.

Methods

Two transversal studies, the first one prior to the start of the IRS campaign and the second one year later. Sampling was carried out by stratified clusters. Malaria infection was measured by means of thick and thin film, and the packed cell volume (PCV) percentage. Data related to ITN use and information regarding IRS were collected. The Pearson's chi-square and logistic regression statistical tests were used to calculate odds ratios (OR)

Results

In the first survey, 168 children were sampled and 433 children in the second one. The prevalence of infection was 40% in 2004, and significantly lower at 21.7% in 2005. PCV was 41% and 39%, respectively. 58% of the children surveyed in 2004 and 44.3% in 2005 had slept under an ITN. 78% of the dwellings studied in 2005 had been sprayed. In the 2005 survey, sleeping without a mosquito net meant a risk of infection 3 times greater than sleeping protected with a net hanged correctly and with no holes (p < 0.05).

Conclusion

IRS and ITNs have proven to be effective control strategies on the island of Bioko. The choice of one or other strategy is, above all, a question of operational feasibility and availability of local resources.