Egg investment in response to helper presence in cooperatively breeding Tibetan ground tits

Life‐history theory predicts a trade‐off between current and future reproduction to maximize lifetime fitness. In cooperatively breeding species, where offspring care is shared between breeders and helpers, helper presence may influence the female breeders’ egg investment, and consequently, survival and future reproductive success. For example, female breeders may reduce egg investment in response to helper presence if this reduction is compensated by helpers during provisioning. Alternatively, female breeders may increase egg investment in response to helper presence if helpers allow the breeders to raise more or higher quality offspring successfully. In the facultatively cooperative‐breeding Tibetan ground tit Pseudopodoces humilis, previous studies found that helpers improve total nestling provisioning rates and fledgling recruitment, but have no apparent effects on the number and body mass of fledglings produced, while breeders with helpers show reduced provisioning rates and higher survival. Here, we investigated whether some of these effects may be explained by female breeders reducing their investment in eggs in response to helper presence. In addition, we investigated whether egg investment is associated with the female breeder's future fitness. Our results showed that helper presence had no effect on the female breeders’ egg investment, and that egg investment was not associated with breeder survival and reproductive success. Our findings suggest that the responses of breeders to helping should be investigated throughout the breeding cycle, because the conclusions regarding the breeders’ adjustment of reproductive investment in response to being helped may depend on which stage of the breeding cycle is considered.     

湖北黄石典型水域超微型真核浮游生物多样性研究

为探讨湖北省黄石市典型水域中超微型真核浮游生物多样性, 采用18S rDNA 扩增片段限制性酶切技术, 结合优势类群测序及相关统计学分析, 对2015年夏季湖北黄石境内营养程度不同的典型水域(长江, 磁湖, 青山湖, 青港湖)中超微型真核浮游生物的群落组成及遗传多样性进行了研究。结果表明, 4个水域的超微型真核浮游生物多样性有明显差异, 多样性随水域营养化程度的增高而降低。优势克隆的测序结果显示, 优势类群藻类包括隐藻、绿藻、甲藻, 真菌包括隐真菌和壶菌, 此外还有纤毛虫及未分类的浮游生物, 隐藻在4个水域中都占优势。不同水域的优势类群差别较大。优势类群的种类随水域营养化程度的增加而减少。群落的多样性与总磷呈极显著的负相关。     

抗克伦特罗单链抗体表达载体的构建与鉴定

为构建克伦特罗(Clenbuterol, CBL)单链抗体(scFv)表达载体, 实现其在大肠杆菌中的表达。以pCANTAB5E- CBL质粒为模板, 扩增CBL的scFv基因, 与pPICZaA载体重组, 然后以重组质粒pPICZaA-scFv为模板扩增scFv及其相连的6 个组氨酸(6×His)片段, 再与载体pBV220连接, 转化大肠杆菌DH5a, 阳性克隆质粒经酶切和PCR鉴定后进行目的片断的序列测定。重组菌经温度诱导表达重组单链抗体, 并通过SDS-PAGE和Western blotting对其鉴定。采用竞争ELISA检测重组单链抗体的抗原结合活性。结果表明重组质粒pBV220-scFv含有插入片段, 与原序列的同源性达99.8%。重组蛋白的分子量接近37 kD, 能够被抗His标签单克隆抗体特异性识别且可被游离的CBL竞争性抑制, IC50值为4.55 ng/mL。这说明我们成功构建了重组质粒pBV220-scFv并实现了其在大肠杆菌中的表达, 为进一步进行CBL免疫法快速检测奠定了一定的基础。     

USP31 acetylation at Lys1264 is essential for its activity and cervical cancer cell growth

Ubiquitin-specific protease 31 (USP31) is a member of deubiquitinase family that is involved in nuclear factor-κB activation and sarcomagenesis.However,little i...     

Grouping Pentylenetetrazol-Induced Epileptic Rats According to Memory Impairment and MicroRNA Expression Profiles in the Hippocampus

Previous studies have demonstrated a close relationship between abnormal regulation of microRNA (miRNA) and various types of diseases, including epilepsy and other neurological disorders of memory. However, the role of miRNA in the memory impairment observed in epilepsy remains unknown. In this study, a model of temporal lobe epilepsy (TLE) was induced via pentylenetetrazol (PTZ) kindling in Sprague-Dawley rats. First, the TLE rats were subjected to Morris water maze to identify those with memory impairment (TLE-MI) compared with TLE control rats (TLE-C), which presented normal memory. Both groups were analyzed to detect dysregulated miRNAs in the hippocampus; four up-regulated miRNAs (miR-34c, miR-374, miR-181a, and miR-let-7c-1) and seven down-regulated miRNAs (miR-1188, miR-770-5p, miR-127-5p, miR-375, miR-331, miR-873-5p, and miR-328a) were found. Some of the dysregulated miRNAs (miR-34c, miR-1188a, miR-328a, and miR-331) were confirmed using qRT-PCR, and their blood expression patterns were identical to those of their counterparts in the rat hippocampus. The targets of these dysregulated miRNAs and other potentially enriched biological signaling pathways were analyzed using bioinformatics. Following these results, the MAPK, apoptosis and hippocampal signaling pathways might be involved in the molecular mechanisms underlying the memory disorders of TLE.     

Central and Midperipheral Corneal Thickness Measured with Scheimpflug Imaging and Optical Coherence Tomography

Purpose

To compare corneal thickness measurements using Pentacam (Oculus, Germany), Sirius (CSO, Italy), Galilei (Ziemer, Switzerland), and RTVue-100 OCT (Optovue Inc., USA).

Methods

Sixty-six eyes of 66 healthy volunteers were enrolled. Three consecutive measurements were performed with each device. The mean value of the three measurements was used for subsequent analysis. Central corneal thickness (CCT), thinnest corneal thickness (TCT), and midperipheral corneal thickness (MPCT; measured at superior, inferior, nasal, and temporal locations with a distance of 1 mm (CT_2mm) or 2.5 mm (CT_5mm) from the corneal apex) were analyzed. Differences and agreement between measurements were assessed using the repeated-measures analysis of variance (ANOVA) and Bland-Altman analyses, respectively.

Results

Statistically significant differences (p<0.001) among the four devices were revealed in CCT, TCT and CT_2mmmeasurements. The CCT, TCT, and CT_2mm values were ranked from the thickest to the thinnest as follows: Galilei>Sirius>Pentacam>RTVue OCT. For these measurements, agreement between measurements by Sirius and Pentacam was good, whereas Galilei overestimated and RTVue underestimated corneal thickness compared to Sirius and Pentacam. As regards CT_5mm measurements, Pentacam provided the largest values, whereas RTVue OCT yielded the smallest values. Agreement of the CT_5mm measurements was good between the Pentacam, Sirius, moderate between Galilei and the other two Scheimpflug systems, and poor between the RTVue OCT and the remaining devices.

Conclusions

The Pentacam and Sirius can be used interchangeably for CCT measurements, while the Galilei and RTVue systematically over- and underestimate CCT, respectively. The three Scheimpflug cameras, but not the RTVue, may be used interchangeably for MPCT measurements.     

我国人乳头瘤病毒（HPV16Z）基因克隆和鉴定

本文作者采用基因克隆手段,以pAt153为载体,从一份山西襄坦宫颈癌高发区宫颈癌患者的手术标本中,成功地克隆到2株与HPV16同源的基因片段。经PstI、KpnI、TaqvI、PvuII等16种限制性内切酶酶谱分析及其部分基因序列的鉴定,证明这是在国内首次克姓到一株分子量约为8．0kb完整的HPV16型全序列DNA及一株分子量为5．4kb的HPV6基因片段。经实验证明：该基因片段E6、E7及部分LI基因丢失,在750单核苷酸处发生变异,产生一新的BamHI酶切位点。该完整的HPV16基因组被命名为HPV16Z,HPV16基因片段被命名为HPV16F。用新分离到的HPV16Z作分子探针,检测襄坦337份宫颈癌及阴道活检标本的HPV16型同源序列的结果显示,慢性阴道炎阳性率为17．28％（14／81）;宫颈炎为11．89％（17／143）;宫颈癌前病变46．81％（22／47）;宫颈癌为72．73％（48／66）。证明山西宫颈癌高发区宫颈癌前病变及宫颈癌组织中主要为HPV16Z感染。     

Bacillus thuringiensis subspecies huazhongensis, serotype H40, isolated from soils in the People's Republic of China

DAI JINGYUAN, YU LING, WANG BO, LUO XIXIA, YU ZINIU AND M.-M. LECADET. 1996. Two isolates (YBt-981 and Ybt-978) of Bacillus thuringiensis were isolated from soil samples from Shanxi and Neimeng provinces in China. The isolates produced small and irregular parasporal inclusions that were not toxic to larvae of Culex fatigans, Culex pipiens, Anopheles sinensis, Aedes aegypti, Spodoptera littoralis, Spodoptera exigua, Plutella xylostella and Bombyx mori . H-antigens from isolates of YBt-981 and YBt-978 differed from those of the known B. thuringiensis H1 to H39 serotypes. YBt-981 and YBt-978 had the same H-antigens and the same biochemical characters. The two isolates were identified as a new serotype designated H40. The name of B. thuringiensis serovar huazhongensis for this new subspecies represented by YBt-981 and YBt-978 is proposed.     

Cloning, expression, purification, and characterization of a novel single-chain variable fragment antibody against the 2-nitrobenzaldehyde derivative of a furaltadone metabolite in Escherichia coli

Furaltadone is an illicit veterinary drug that shows toxic, carcinogenic, and mutagenic effects, as does its metabolite 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ)(1). Recombinant antibodies with desirable affinity and specificity that can replace polyclonal or monoclonal antibodies are important factors for effective AMOZ immunoassays. In the present study, a novel single-chain variable fragment (scFv) antibody against the 2-nitrobenzaldehyde derivative of AMOZ (NPAMOZ) was prepared and characterized. The scFv gene was cloned into the pET-22b(+) expression vector, and 6His-tagged scFv antibodies expressed as inclusion bodies in Escherichia coli BL21 (DE3), which were then purified by nickel nitrilotriacetic acid column chromatography. Characterization of the target protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and a novel indirect competitive chemiluminescence enzyme immunoassay (icCLEIA) showed that the scFv antibody was ～27kDa and exhibited HRP-anti-His-tag antibody-recognized activity. The final purity, yield and mg of this scFv antibody after ultrafiltration concentration were 97%, 20% and 29.1mg, respectively. The icCLEIA indicated that the antibody competitively combined with NPAMOZ, exhibiting an IC(50) value of 1.46±0.01 ng/ml (n=6). Cross-reactivity studies revealed that the antibody showed desirable specificity to NPAMOZ and little reactivity to analogs except the parent furaltadone. In summary, these findings suggested that the prepared recombinant scFv antibody can be used for future immunoassay screening for AMOZ.     

Production and Characterization of a Single-Chain Fv Antibody–Alkaline Phosphatase Fusion Protein Specific for Clenbuterol

The production and characterization of an anti-clenbuterol single-chain Fv antibody (CBLscFv)–bacterial alkaline phosphatase (AP) fusion protein are described. The CBLscFv and the phoA gene of Escherichia coli strain K12 chromosomal DNA were cloned by PCR and sequentially inserted into the expression vector pBV220 to express the CBLscFv–AP fusion protein in E. coli strain BL21(DE3)pLysS. SDS–PAGE and western blot analyses revealed that the fusion protein showed a molecular weight of 73 kDa and bound with the antibacterial AP monoclonal antibody. Determination of enzymatic activity indicated that k _cat and K _m values of the fusion protein were 113.60 s⁻¹ and 29.82 μM, respectively. Competitive direct enzyme-linked immunosorbent assay based on the obtained fusion protein indicated that the average concentration required for 50% inhibition of binding (IC₅₀) and the limit of detection for CBL were 4.74 ± 0.003 (n = 3) and 0.54 ± 0.004 (n = 3) μg/l, respectively, and the linear response range extended from 1.13 to 69.68 μg/l. Cross-reactivity studies showed that the fusion protein did not cross-react with CBL analogs. The present findings indicate that the production of the CBLscFv–AP fusion protein in E. coli strain BL21(DE3)pLysS is feasible and suggest that it could be further used to develop a one-step ELISA for the specific detection of CBL.