Bovine mammary dendritic cells: a heterogeneous population, distinct from macrophages and similar in phenotype to afferent lymph veiled cells

Dendritic cells (DC) are a heterogeneous population of professional antigen presenting cells and are potent stimulators of na?ve T-cells. However, there is little previous research describing DC in bovine mammary tissue, primarily because of the difficulty distinguishing these cells from macrophages, which possess a similar phenotype. Using immunohistofluorescence and a combination of markers (MHC-II, CD205, CD11c), DC were localized in the bovine mammary gland and supramammary lymph node. In mammary tissue DC were found within the alveolar epithelium and within the intralobular connective tissue. In the lymph node DC were found on the periphery of B-cell areas, in the cortex, and among T-cells in the paracortex and medulla. DC in mammary parenchyma and supramammary lymph nodes were quantified and further characterized using flow cytometry. DC were CD11c(hi), CD14(lo) cells that expressed MHC-II and CD205. DC could be distinguished from macrophages based on their low CD14 expression. This research provides a better understanding of mammary gland immunology, while potentially aiding in the targeting of antigens to mucosal DC for vaccine development.     

棉花gDNA体细胞染色体FISH技术

介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ,简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程,并着重分析和讨论了影响试验结果的关键因素,包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉;探针和封阻均采用ｇＤＮＡ,材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等）,分别交互使用。试验结果比较理想,获得良好的ＦＩＳＨ片子,而且重复性好。     

通过饲喂浓缩玉米酒糟可溶物和鱼油提高牛奶中共轭亚油酸含量

选用12头荷斯坦奶牛随机分组饲喂4种日粮中的一种,采用重复4×4拉丁方设计,每期试验4周,研究饲喂鱼油(FO)、浓缩玉米酒糟可溶物(CDS)作为额外添加亚油酸或两者同时添加时对奶牛泌乳性能的影响。日粮中不添加FO和添加0.5%FO,不添加CDS和添加10?S,采用2×2因子试验设计。采用全混合日粮(TMR)自由采食,粗料和精料比例为55∶45(以干物质计),含16.2%粗蛋白质。对照组、FO组、CDS组和FOCDS组粗脂肪分别为2.86%、3.22%、4.77%和5.02%。FO组、CDS组及两者同时添加的日粮分别与不添加FO组和不添加CDS的日粮组相比,对干物质采食量、饲料转化率、体增重和体况评分无显著影响。各日粮组乳蛋白产量(33.3 kg/d)、能量校正乳、乳蛋白、乳糖和乳中尿素氮相近。与不饲喂FO和CDS相比,饲喂FO和CDS降低乳脂百分率(4个试验组分别为3.85%、3.39%、3.33%和3.12%)和乳脂产量。饲喂FO和CDS提高trans-11 C18∶1(十八碳烯酸),cis-9,trans-11共轭亚油酸(每100 g脂肪酸中CLA含量分别为0.52、0.90、1.11和1.52 g)和trans-10 cis-12 CLA浓度(100 g脂肪酸中分别为0.07、0.14、0.13和0.16 g)。FO和CDS对cis-9,trans-11 CLA的影响没有互作作用,但对十八碳烯酸趋于提高方面存在互作作用。添加CDS可提高trans-10 C18∶1,饲喂FO和CDS血浆中十八碳烯酸与cis-9,trans-11 CLA比率高于乳脂中的比率,这表明饲喂FO和CDS促进乳腺组织中十八碳烯酸合成cis-9,trans-11 CLA。日粮中添加占干物质0.5%的鱼油和富含C18∶2 n-6脂肪酸的CDS提高牛奶中CLA含量,降低乳脂百分率。     

Homologies in leaf form inferred from KNOXI gene expression during development

KNOTTEDI-like homeobox (KNOXI) genes regulate development of the leaf from the shoot apical meristem (SAM) and may regulate leaf form. We examined KNOXI expression in SAMs of various vascular plants and found that KNOXI expression correlated with complex leaf primordia. However, complex primordia may mature into simple leaves. Therefore, not all simple leaves develop similarly, and final leaf morphology may not be an adequate predictor of homology.     

棉花花粉中高效转录U6启动子的克隆及功能分析

【目的】以棉花品种新海16基因组DNA为模板,克隆海岛棉（Gossypium barbadense L.）GbU6启动子,筛选出能在棉花生殖细胞中（花粉）高效转录的GbU6启动子,为利用基因组编辑技术进行棉花分子育种奠定重要基础。【方法】采用两轮PCR方法克隆完整的GbU6启动子。根据网上公布的棉花基因组数据库序列,利用Primer 5.0软件设计5对引物,进行第一轮能覆盖完整GbU6启动子的PCR扩增,产物克隆测序正确后,再利用transfer PCR法将GbU6启动子精确亚克隆到CRISPR/Cas9基因组编辑载体中;第二轮扩增产物测序正确后,运用DNAMAN软件对克隆成功的5个GbU6启动子序列中具有转录功能的必要元件进行分析;然后以pBI101质粒DNA为模板,PCR扩增并克隆GUS报告基因,经酶切鉴定、测序正确后,用BbsⅠ酶切GUS,连入经同样酶切后的5个GbU6启动子对应的CRISPR/Cas9基因组编辑载体中,转化、酶切鉴定,获得对应的5种GbU6::GUS的表达载体。将含有CaMV35S启动子驱动的GUS表达载体作为棉花花粉瞬时转化的阳性对照,以上述6种表达载体DNA为模板,采用高保真酶的PCR扩增法获得高浓度DNA片段,利用基因枪轰击法将5种GbU6::GUS和阳性对照的DNA片段分别转化棉花花粉,并进行GUS染色,最后用体式显微镜观察染色情况。每个启动子的基因枪转化重复3次,最后根据染色深浅筛选出在棉花生殖细胞中高效转录的GbU6启动子。【结果】经两轮PCR扩增后获得5种GbU6启动子,启动子长度分别为1 166、1 119、1 134、1 214和1 176 bp,并构建获得了相应的5种CRISPR/Cas9基因组编辑载体;对拟南芥和克隆的5个GbU6启动子序列进行序列比对,结果表明,GbU6启动子区与拟南芥U6启动子一样,也含有比较保守的-60 bp位置的USE元件和-30 bp位置的TATA框,而且这两个元件之间的距离也非常固定;用GbU6::GUS的DNA片段瞬时转化棉花花粉,发现基因枪瞬时转化的3次重复结果显示克隆得到的5个GbU6启动子有4个能驱动GUS在棉花花粉中表达,棉花花粉被染成蓝色。其中GbU6-5P::GUS的染色相对较深,接近于CaMV35S启动子。【结论】成功克隆了棉花生殖细胞中高效转录的GbU6启动子,为构建棉花CRISPR/Cas9基因组编辑载体系统提供了有效的启动子。     

Structure and regeneration status of mangrove patches along the estuarine and coastal stretches of Kerala,India

This study presents the structural characteristics and regeneration potential of mangrove patches in the estuarine and coastal areas of Kerala, a tropical maritime state in India. Field surveys were carried out at 46 selected sites during August 2015 to May 2016. In each site, the vegetative structure and regeneration status were assessed using the quadrat method. Altogether 219 quadrates were laid out and a total of 13 true mangrove species, belonging to 5 families and 8 genera, were recorded. The total tree density and stand basal area of the study region was1678.08/ha and 20.33 m^2/ha respectively. The low basal areas indicate the reduced structural development in mangroves. Of the 13 tree species, Avicennia constitutes 56%of the total Important Value Index(IVI) and Avicennia officinalis represents 41% of the IVI in Kerala, followed by Avicennia marina(15%), Rhizophora mucronata(15%),Sonneratia alba(8%) Rhizophora apiculata(7%) and Excoecaria agallocha(7%). The diameter at breast height(DBH) in the study area revealed that 47% of the tree species came under the 1–10 cm DBH class. Total sapling and seedling density in Kerala was 2238.35 and 3232.42 individuals/ha respectively. Density of young plants(seedlings ? saplings) was only 31% greater of tree density and varied from 3–63%, which indicates poor regeneration potential. The Maturity index value(MIV) and complexity index(Ic) value of mangroves were 18.30 and 109.81 respectively. However, the low Ic value(\ 10) observed in seven out of ten coastal districts indicated poor structural development of mangroves in Kerala. Therefore, locationspecific conservation and management measures, guided by the knowledge on spatial distribution and habitat requirements of mangrove varieties should be taken to preserve the mangrove diversity of Kerala.     

Prevalence of rabies antibodies in street and household dogs in Chandigarh,India

The present study reports the first survey for the detection of antirabies antibodies in street and household dogs in India. We aimed to check the efficacy of control programs for the disease in the Union territory of Chandigarh. The serum samples were collected from 100 street and 50 household dogs and tested for the presence of antirabies antibodies by ELISA. As per WHO criteria, a titre of >0.5 IU/ml of antirabies antibody in serum samples was taken as the protective level. Protective antirabies antibody titre was found only in 1% of the street dogs and 16% of the pet dogs. The awareness among the pet dog owners about the antirabies vaccination schedule was low as 18% did not know the vaccination status of the dog and 66% had got the initial immunization done with only three doses and annual boosters were not given. A National Rabies Elimination Program needs to be launched as a collaborative venture by both medical and veterinary practitioners to curb this deadly disease. Also periodic surveys to test the status of antirabies IgG among dogs need to be carried out to ascertain the attainment of WHO protective levels     

不同颜色薄膜对设施环境及甜椒生长发育和产量品质的影响

以无色透明膜为对照（CK）,在相近光照强度下,研究了紫色膜、蓝色膜和红色膜覆盖对设施环境及甜椒生长发育和产量品质的影响。结果表明：不同颜色薄膜覆盖对设施内环境因子的影响不大;红色膜覆盖甜椒茎粗最大,株高较小,而蓝色膜覆盖甜椒茎粗最小,株高较大;蓝色膜覆盖延迟了甜椒开花时间,红色膜覆盖则使甜椒开花期提前;甜椒单株产量和单株果数均以蓝色膜覆盖最低,红色膜覆盖最高,单果重以紫色膜处理最高;蓝色膜覆盖提高了甜椒果实Vc和可溶性蛋白含量,果实可溶性糖和游离氨基酸含量则以红色膜处理最高。     

Double-stranded RNA: distribution and analysis among isolates of Rhizoctonia solani AG-2 to -13

The occurrence, distribution, and genetic relatedness of double-stranded RNA (dsRNA) components from 36 isolates of Rhizoctonia solani belonging to nine anastomosis groups (AGs) were studied using electrophoretic analysis and RNA–RNA blot hybridization. DsRNA was consistently detected in all 36 isolates. The size of the dsRNA components varied considerably, ranging from 0·74 to 23 kb. Two thirds of isolates possessed different size dsRNA components. Only two of the isolates had small size dsRNA between 0·5 and 1·0 kb. The biotin-labelled dsRNA probes provided the sensitivity and specificity required to study genetic relationships of dsRNA. As little as 10 pg of 'hybridizing' dsRNA could be detected using biotin-labelled total dsRNA with no detectable nonspecific-hybridization. Results from several dot-spot as well as RNA–RNA gel hybridization experiments revealed considerable sequence heterogeneity among dsRNA components within each isolate or isolates from the same AG.