Characterization and allelic variation of the transporters associated with antigen processing (TAP) genes in the domestic dog (Canis lupus familiaris)

The function of the transporters associated with antigen processing (TAP) complex is to shuttle antigenic peptides from the cytosol to the endoplasmic reticulum to load MHC class I molecules for CD8⁺ T-cell immunosurveillance. Here we report the promoter and coding regions of the canine TAP1 and TAP2 genes, which encode the homologous subunits forming the TAP heterodimer. By sampling genetically divergent breeds, polymorphisms in both genes were identified, although there were few amino acid differences between alleles. Splice variants were also found. When aligned to TAP genes of other species, functional regions appeared conserved, and upon phylogenetic analysis, canine sequences segregated appropriately with their orthologs. Transfer of the canine TAP2 gene into a murine TAP2-defective cell line rescued surface MHC class I expression, confirming exporter function. This data should prove useful in investigating the association of specific TAP defects or alleles with immunity to intracellular pathogens and cancer in dogs.     

泪小管鼻侧断端位置分区的临床应用

目的：观察泪小管断裂后鼻侧断端位置与泪阜和内眦韧带的关系及其在泪小管断裂吻合术中的应用。

方法：选取外伤性下泪小管断裂患者65例65眼,测量泪小管泪点至颞侧断端的距离,直视下寻找鼻侧断端,量化分析鼻侧断端与泪阜、内眦韧带的关系。

结果：本组患者颞侧断端距离≤6mm者39眼,其中鼻侧断端位于泪阜区者35眼; 颞侧断端距离>6mm者26眼,其中鼻侧断端位于内眦韧带区者24眼。泪阜区鼻侧断端距离泪阜下缘黏膜的垂直距离为2.1±0.7mm,内眦韧带区鼻侧断端距离皮肤面的垂直距离为3.5±1.2mm。

结论：泪小管断裂后将鼻侧断端位置分为泪阜区和内眦韧带区,有助于寻找鼻侧断端位置。     

反式二羟环氧苯并（a）芘相关新基因brg的全长克隆

背景与目的:在前期研究中发现,16HBE-C细胞表达变化的基因中有未知基因参与,因其与BPDE有关,所以命名为brg(anti-BPDErelatedgene)基因。本研究应用cDNA末端快速扩增法(rapidamplificationofcDNAends,RACE)扩增此未知基因的全长,以进行下一步的基因功能研究。材料与方法:应用cDNA末端快速扩增法(RACE技术),对3'和5'端分别设计了梯度PCR,巢式PCR等优化程序,以获得特异的产物,然后对特异产物直接测序,将测序结果进行生物信息学分析,基因拼接等获得新基因brg全长。结果:3'和5'端均成功获得特异性产物,3'RACE测序为394bp,有明显的PolyA加尾信号,有编码区的终止密码子;5'RACE测序为1017bp,有起始密码子ATG,其中197bp为3'和5'全长共有序列。将3'和5'序列拼接,结果全长1214bp,生物信息学分析brg基因阅读框1032bp,编码344个氨基酸。结论:所得结果与设计一致,说明所采用的技术路线是简便可行的,brg基因可能在反式二羟环氧苯并(a)芘的致癌机制中起重要作用。     

Influence of orthodontic premolar extraction therapy on the eruption of the third molars: A systematic review of the literature

Background

Through a systematic literature review, the authors assess the effect of premolar extractions on third-molar (M3) eruption considering eruption rate, retromolar space, and molar angulation.

Molecular and immunological characterization of subtilisin like serine protease, a major allergen of Curvularia lunata

Serine protease from numerous sources have been identified and characterized as major allergens. The present study aimed to clone, express and characterize a serine protease from Curvularia lunata. cDNA library screening identified partial protease clones. A clone showed significant homology to subtilisin like serine proteases from Aspergillus and Penicillium species. Full length sequence was generated by RACE PCR, subcloned in pET vector, protein expressed in Escherichia coli and purified from inclusion bodies yielding 0.5 mg/L of culture. Bioinformatic analysis identified serine protease motifs of subtilase family, catalytic triad and N-glycosylation sites on the primary sequence. The protein resolved at 54-kDa on SDS-PAGE and was recognized as a major allergen on immunoblot with 13/16 C. lunata sensitive patients’ sera in ELISA and immunoblot. Recombinant protein reacted with rabbit polyclonal antibodies against alkaline serine proteases from C. lunata. Recombinant protein required 50-56 ng of same protein for 50% inhibition of IgE binding in competitive ELISA. In addition, 13 of 16 patients’ samples showed significant basophil histamine release upon stimulation with purified recombinant protein. In conclusion, a 54 kDa major allergen of C. lunata was cloned, expressed, characterized and showed biological activity. It has potential to be used in molecule based approach for allergy diagnosis and therapy.