Poinsettia's wild ancestor in the Mexican dry tropics: Historical, genetic, and environmental evidence

? Premise of the study: The poinsettia (Euphorbia pulcherrima) is the world's most economically important potted plant, but despite its preeminence it is not clear which wild populations are ancestral to the varieties cultivated around the world. Tradition holds that the U.S. envoy to Mexico J. R. Poinsett collected the progenitors of the over 300 varieties in global cultivation on an 1828 excursion to northern Guerrero State, Mexico. It is unknown whether the contemporary cultivars are descended from plants from Guerrero or whether germplasm from other parts of poinsettia's 2000 km long distribution entered into cultivation during the nearly 200 yr of subsequent poinsettia horticulture. ? Methods: To identify the wild populations that likely gave rise to the cultivars and test this historical account, we sequenced plastid and nuclear DNA regions and modeled poinsettia's potential distribution. ? Key results: The combination of nuclear and plastid haplotypes characterizing cultivars was found only in northern Guerrero. Distribution modeling indicated that suitable habitat conditions for wild poinsettias are present in this area, consistent with their likely wild status. ? Conclusions: Our data pinpoint the area of northern Guerrero as the cultivated poinsettia's probable ancestral region, congruent with the traditional account attributing the original collections to Poinsett. Abundant genetic variation likely offers raw material for improving the many shortcomings of cultivars, including vulnerability to cold, stem breakage, and pathogens such as Pythium and Phytophthora. However, genetic differences between populations make conservation of all of poinsettia's diversity difficult.     

Surface proteins from Lactobacillus kefir antagonize in vitro cytotoxic effect of Clostridium difficile toxins

In this work, the ability of S-layer proteins from kefir-isolated Lactobacillus kefir strains to antagonize the cytophatic effects of toxins from Clostridium difficile (TcdA and TcdB) on eukaryotic cells in vitro was tested by cell detachment assay. S-layer proteins from eight different L. kefir strains were able to inhibit the damage induced by C. difficile spent culture supernatant to Vero cells. Besides, same protective effect was observed by F-actin network staining. S-layer proteins from aggregating L. kefir strains (CIDCA 83115, 8321, 8345 and 8348) showed a higher inhibitory ability than those belonging to non-aggregating ones (CIDCA 83111, 83113, JCM 5818 and ATCC 8007), suggesting that differences in the structure could be related to the ability to antagonize the effect of clostridial toxins. Similar results were obtained using purified TcdA and TcdB. Protective effect was not affected by proteases inhibitors or heat treatment, thus indicating that proteolytic activity is not involved. Only preincubation with specific anti-S-layer antibodies significantly reduced the inhibitory effect of S-layer proteins, suggesting that this could be attributed to a direct interaction between clostridial toxins and L. kefir S-layer protein. Interestingly, the interaction of toxins with S-layer carrying bacteria was observed by dot blot and fluorescence microscopy with specific anti-TcdA or anti-TcdB antibodies, although L. kefir cells did not show protective effects. We hypothesize that the interaction between clostridial toxins and soluble S-layer molecules is different from the interaction with S-layer on the surface of the bacteria thus leading a different ability to antagonize cytotoxic effect. This is the first report showing the ability of S-layer proteins from kefir lactobacilli to antagonize biological effects of bacterial toxins. These results encourage further research on the role of bacterial surface molecules to the probiotic properties of L. kefir and could contribute to strain selection with potential therapeutic or prophylactic benefits towards CDAD.     

海藻酸微囊替代DMSO和FBS的STO细胞冷冻保存研究

目的:改进现有的细胞冷冻保存方法,建立一个不含二甲基亚砜(DMSO)和血清(FBS)的高效冷冻保存方法,为细胞治疗等临床实践提供优质细胞.方法:海藻酸微囊包埋鼠胚成纤维细胞(STO细胞)后用不含DMSO和FBS的冷冻保存液进行冷冻保存.设四个对照组:添加10％DMSO和20％FBS的组、仅添加10％DMSO的组、仅添加20％FBS、DMSO和FBS均不添加组.在冷冻前后对各实验组细胞用台盼兰染色,进行细胞计数,计算细胞存活率,同时利用溴乙锭的二聚物(EthD)、钙黄绿素-AM(Calcein-AM)进行染色观察细胞的形态,且进一步验证细胞存活率;解冻复苏后用MTT法评估细胞的增殖速度和生长活力.结果:冷冻保存30天后对各组的细胞数量、细胞存活率、细胞形态和解冻复苏后细胞的生长活力进行比较发现,海藻酸微囊包埋冷冻组的细胞数、细胞存活率、细胞形态和生长活力均与添加DMSO和FBS的组之间无显著性差异,而与其它三个对照组呈显著性差异.结论:使用海藻酸微囊替代DMSO和FBS保存STO细胞,能有效的维持细胞形态、数量、存活率,同时不影响细胞的生长活力,从而建立了一个不含DMSO和FBS的高效冷冻保存方法.     

H-FABP基因的多态性和营养因素对猪肉质的影响

遗传和营养因素都能影响猪肉的品质。但是, 到目前为止同时研究遗传和营养因素对肉质影响的报道很少。在本研究中, 136头PIC5系杂交猪, 体重65 kg, 被随机分成4组, 各组分别给予不同日粮。在饲养35 d、体重大约90 kg时统一屠宰并且进行肉质测定、H-FABP基因分型及其与肉质性状的关联分析。结果表明: (1)所采用的3种日粮对肉色、屠宰后24 h的pH、肌内脂肪和肌肉蛋白含量有极显著的影响; (2)H-FABP基因型对肌内脂肪和肌肉蛋白含量存在极显著的影响; (3)H-FABP基因多态性和营养因素的交互作用对pH 和肌内脂肪含量均有显著的影响, 对照组的AA基因型具有最高pH值, 高维生素E组的AA基因型具有最高肌内脂肪值。实验结果提示在关于猪肉质的育种和生产过程中应该同时考虑营养因素和遗传因素。     

披针叶黄华的界定

通过对标本的比较研究及广泛的野外考察,界定了披针叶黄华,承认了一个种级名称Thermopsis lupinoides（L．）Link．并组合了9个新异名。     

MDR1基因单靶点双荧光素酶报告基因系统的建立

目的通过构建以MDR1启动子为启动序列的双荧光素酶报告基因系统并进行活性分析,为MDR1基因表达的单靶点调控研究和逆转剂的筛选提供一种有效的方法。方法从HCT-8细胞中提取DNA并克隆含有MDR1基因启动子中Y—box序列。将该序列重组到萤光素酶报告基因载体pGL-3．Basic的启动区域中,从而构建报告基因载体pGL-MDR1。将pGL-MDR1和pRL-TK载体共转染到HCT-8和HCT-8／VCR细胞中。通过调节不同载体的比例来优化转染效率。利用MDRI基因激活剂（热诱导）和抑制剂（EGCG）等处理来分析其启动转录活性受外界因素的影响。结果通过直接测序法验证了pGL-MDR1含有MDR1基因启动子Y—box序列且没有出现碱基突变。在pGL-MDR1和pRL-TK的转染比例为5：5时,转染效率最高并具有最高的萤光素酶活性。通过MDR1基因激活处理后表现为时间依赖性地激活MDR1基因的表达,而MDR1基因抑制剂的作用则相反。结论MDR1启动子为启动序列的双荧光素酶报告基因系统建立成功。该系统不但可以用于研究活体生物发光成像和MDRI基因表达的机理,而且可用于单靶点的多药耐药抑制剂的筛选。     

Clustering cancer gene expression data: a comparative study

Background  

The use of clustering methods for the discovery of cancer subtypes has drawn a great deal of attention in the scientific community. While bioinformaticians have proposed new clustering methods that take advantage of characteristics of the gene expression data, the medical community has a preference for using "classic" clustering methods. There have been no studies thus far performing a large-scale evaluation of different clustering methods in this context.     

Purification and Characterization of Hieronymain III. Comparison with Other Proteases Previously Isolated from Bromelia hieronymi Mez

A new proteolytic enzyme, named hieronymain III, has been purified by ion-exchange chromatography from unripe fruits of Bromelia hieronymi Mez. The new peptidase belongs to the cysteine catalytic type, as well as hieronymain I and II, the other two peptidases previously isolated from this species. Hieronymain III showed optimum alkaline pH range (8.6–9.3) and the molecular mass (MALDI-TOF) was 23713 Da. The N-terminal sequence (AVPQSIDWRRYGAVTTSRNQG) exhibited a higher percentage identity with hieronymain II (93%) than with hieronymain I (71%). The three peptidases showed notable differences on synthetic substrates degradation: whereas hieronymain III was the only one able to hidrolyze Z-Arg-Arg-p-nitroanilide, hieronymain I and II could degrade Z-Phe-Arg-p-nitroanilide; on the other hand, PFLNA was only split by hieronymain I. Finally, the three proteases showed different preferences on N-α-CBZ-p-nitrophenyl aminoacid ester substrates. From a biotechnological point of view, cleavage specificity differences are significant enough to use these enzymes as potential tools in that area.     

Indices of anti‐dengue immunoglobulin G subclasses in adult Mexican patients with febrile and hemorrhagic dengue in the acute phase

Heterologous secondary infections are at increased risk of developing dengue hemorrhagic fever (DHF) because of antibody‐dependent enhancement (ADE). IgG subclasses can fix and activate complement and bind to Fc? receptors. These factors may also play an important role in the development of ADE and thus in the pathogenesis of DHF. The aim of this study was to analyze the indices of anti‐dengue IgG subclasses in adult patients with febrile and hemorrhagic dengue in the acute phase. In 2013, 129 patients with dengue fever (DF) and 57 with DHF in Veracruz, Mexico were recruited for this study and anti‐dengue IgM and IgG determined by capture ELISA. Anti‐dengue IgG subclasses were detected by indirect ELISA. Anti‐dengue IgG2 and IgG3 subclasses were detected in patients with dengue. IgG1 increased significantly in the sera of patients with both primary and secondary infections and DHF, but was higher in patients with secondary infections. The IgG4 subclass index was significantly higher in the sera of patients with DHF than in that of those with DF, who were in the early and late acute phase of both primary and secondary infection. In conclusion, indices of subclasses IgG1 and IgG4 were higher in patients with DHF.