Light microscopic visualization of peroxisomes and plasmalogens in first trimester chorionic villi

Peroxisomes and their metabolites the plasmalogens were visualized in chorionic villi by quick and simple procedures. Villi were stained and mounted in toto, and can also be embedded. These methods may contribute to early prenatal diagnosis of peroxisomal disorders (Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum's disease a.o.).     

Immunocytochemical localization of peroxisomal β-oxidation enzymes in human fetal liver

In the majority of congenital peroxisomal disorders, β-oxidation of very long chain fatty acids is deficient. We have investigated the appearance and localization of the three peroxisomal β-oxidation enzymes in normal fetal liver (fertilization age between 5 and 18 weeks) with protein A- gold immunocytochemistry and silver enhancement for light microscopic visualization. With specificity-tested polyclonal antibodies, acyl-CoA-oxidase, bifunctiooal enzyme, and 3-oxoacyl-CoA thiolase were localized in the peroxisomes of the parenchymal cells, which appear as brown or black granules. In the youngest specimen, no immunopositive reaction was obtained. A weak reaction with anti-thiolase was obtained at the age of 6–7 weeks. At a fertilization age of 8 weeks, peroxisomes could be distinctly visualized after immunostaining for all three enzymes. From a staining series with anti-thiolase on simultaneously treated slides, it appears that the amount of antigen per peroxisome and the organelle size increase between the seventh and eighteenth weeks. These data should enable a more specific diagnosis in fetal liver biopsies from pregnancies at risk and after termination of pregnancy.     

Immunoblot detection of enzyme proteins of peroxisomal β-oxidation in fibroblasts,amniocytes, and chorionic villous cells. Possible marker for prenatal diagnosis of Zellweger's syndrome

The amounts of enzyme proteins of peroxisomal β-oxidation in fibroblasts and chorionic villous cells from infants with Zellweger syndrome and in fibroblasts, amniocytes, and chorionic villi from healthy controls were measured by immunoblot analysis. Immunoreactive proteins of peroxisomal acyl-CoA oxidase and 3-ketoacyl-CoA thiolase were absent in fibroblasts and chorionic villous cells from the patients, yet these enzyme proteins were present in fibroblasts, cultured amniocytes, and chorionic villi from the normal controls. These results show that immunoblot analysis of peroxisomal β-oxidation enzymes in amniocytes and chorionic villous cells is of potential value for the prenatal detection of Zellweger syndrome.     

Meta-networks for the study of biogeographical traits in ecological networks: the Mexican hummingbird-plant assemblage

Recent studies on ecological networks have quantified the contribution of ecological, historical, and evolutionary factors on the structure of local communities of interacting species. However, the influence of species’ biogeographical traits, such as migratory habits or phylogeographical history, on ecological networks is poorly understood. Meta-networks, i.e., networks that cover large spatial extensions and include species not co-occurring locally, enable us to investigate mechanisms that operate at larger spatial scales such as migratory patterns or phylogeographical distributions, as well as indirect relationships among species through shared partners. Using a meta-network of hummingbird-plant interaction across Mexico, we illustrate the usefulness of this approach by investigating (1) how biogeographical and morphological factors associate with observed interactions and (2) how species-specific biogeographical characteristics associate with species’ network roles. Our results show that all studied hummingbird and plant species in the meta-network were interrelated, either directly or through shared partners. The meta-network was structured into modules, resulting from hummingbirds and plants interacting preferentially with subsets of species, which differed in biogeographical, and, to a lesser extent, morphological traits. Furthermore, migrants and hummingbirds from Nearctic, Transition, and widespread regions had a higher topological importance in the meta-network. Our study illustrates how meta-networks may contribute to our current knowledge on species’ biogeographical traits and biotic interactions, providing a perspective complementary to local-scale networks.     

Fetal liver alanine: Glyoxylate aminotransferase and the prenatal diagnosis of primary hyperoxaluria type 1

Primary hyperoxaluria type 1 (PH1) is caused by a deficiency of the hepatic peroxisomal enzyme alanine: glyoxylate aminotransferase (AGT, EC 2.6.1.44) (Danpure and Jennings, FEBS Lett., 201 , 20–24, 1986). The activity of AGT has been measured in fetal livers of gestational age 14–21 weeks. Activity increases up to 17 weeks and then levels off between 17 and 21 weeks. At this time, the mean AGT activity is about 30 per cent of the mean normal postnatal level. As in adult liver, the AGT enzyme activity and the AGT immunoreactive protein are peroxisomal. Prenatal diagnosis has been performed by measuring AGT enzyme activity and immunoreactive AGT protein on liver biopsies from two fetuses at risk for primary hyperoxaluria type 1. One was unaffected and one was affected.     

The continuing conundrum of the LEA proteins

Research into late embryogenesis abundant (LEA) proteins has been ongoing for more than 20 years but, although there is a strong association of LEA proteins with abiotic stress tolerance particularly dehydration and cold stress, for most of that time, their function has been entirely obscure. After their initial discovery in plant seeds, three major groups (numbered 1, 2 and 3) of LEA proteins have been described in a range of different plants and plant tissues. Homologues of groups 1 and 3 proteins have also been found in bacteria and in certain invertebrates. In this review, we present some new data, survey the biochemistry, biophysics and bioinformatics of the LEA proteins and highlight several possible functions. These include roles as antioxidants and as membrane and protein stabilisers during water stress, either by direct interaction or by acting as molecular shields. Along with other hydrophilic proteins and compatible solutes, LEA proteins might also serve as “space fillers” to prevent cellular collapse at low water activities. This multifunctional capacity of the LEA proteins is probably attributable in part to their structural plasticity, as they are largely lacking in secondary structure in the fully hydrated state, but can become more folded during water stress and/or through association with membrane surfaces. The challenge now facing researchers investigating these enigmatic proteins is to make sense of the various in vitro defined functions in the living cell: Are the LEA proteins truly multi-talented, or are they still just misunderstood?     

Molecular analysis of genomic DNA allows rapid,and accurate,prenatal diagnosis of peroxisomal D-bifunctional protein deficiency

Prenatal diagnosis was requested for a couple with a previous child affected by the peroxisomal disorder D-bifunctional protein deficiency. Prior analysis of the D-bifunctional protein cDNA sequence from the propositus had shown that it was missing 22 bp. This was subsequently attributed to a point mutation in the intron 5 donor site (IVS5+1G>C) of the D-bifunctional protein gene. Consistent with parental consanguinity, the patient was shown to be homozygous for this mutation, which is associated with loss of a Hph 1 restriction site in the genomic sequence. Prenatal testing of the fetus using genomic DNA isolated from uncultured amniocytes indicated that both alleles of the D-bifunctional protein had the IVS5+1G>C substitution. The peroxisomal defect was later confirmed biochemically using cultured amniocytes, which were found to have elevated levels of very long chain fatty acids (VLCFA). This is the first report of prenatal diagnosis of D-bifunctional protein deficiency using molecular analysis of genomic DNA. Copyright © 2002 John Wiley & Sons, Ltd.     

Prenatal diagnosis of inborn errors in peroxisomal β-oxidation

In recent years, an increasing number of inherited diseases in man have been recognized in which there is an impairment in the peroxisomal β-oxidation of very-long-chain fatty acids. In general, these disorders are associated with severe neurological and physical abnormalities and death within the first years of life. In this paper we describe our experience with regard to the prenatal diagnosis of a number of different inborn errors of peroxisomal β-oxidation. Eleven pregnancies at risk were monitored by measuring very-long-chain fatty acid levels as well as very-long-chain fatty acid β-oxidation in cultured chorionic villous fibroblasts and/or amniotic fluid cells. Five affected fetuses were identified. It is concluded that prenatal diagnosis in this group of diseases can be done reliably using cultured chorionic villous fibroblasts or amniotic fluid cells.     

Responses of wheat seedlings to cadmium, mercury and trichlorobenzene stresses

The molecular response of wheat (Triticum aestivum L.,cv.Yangmai 10) seedlings to heavy metal (Cd,Hg) and 1,2,4-trichlorobenzene (TCB) stresses were examined by two-dimensional gel electrophoresis,image analysis,and peptide mass fingerprinting.The results showed inhibitions of root and shoot growth by Cd,Hg and TCB.These stresses led to water deficit and lipid phosphorylation in the seedling which also promoted protein phophorylation in the leaves.Hg stress inhibited protein synthesis while Cd and TCB stresses induced or up-regulated more proteins in the leaves.Most of these induced proteins played important roles in the biochemical reactions involved in tolerance of wheat to Cd and TCB stresses.The primary functions of Cd- and TCB-induced proteins included methionine metabolism,Rubisco modification,protein phosphorylation regulation,protein configuration protection,H+ transmembrane transportation and also the synthesis of ethylene defense substances and cell wall compounds.     

Possible role of extracellular vesicles in exogenous chemical exposure-associated health concern

Extracellular vesicles(EVs) are mediators of intercellular message communication by transferring lipids, proteins, genetic materials, and are therefore involved in a wide spectrum of physiological or pathological processes. Exogenous chemicals exposure may affect the function of EVs at the biogenesis, secretion, transport or reception stage, which could further result in abnormal information communication between cells, and thus induce toxicity and diseases. This review briefly explores the role of EVs in chemical exposure-associated health concern and progressive diseases.     

红球菌跨膜运输荧蒽的差异膜蛋白分析

提取不同时间荧蒽诱导下红球菌BAP-1分泌的膜蛋白,利用同位素标记相对和绝对定量技术（iTRAQ技术）结合液相二级质谱对差异膜蛋白进行聚类以及生物信息学分析.旨在从蛋白质层面上研究红球菌BAP-1跨膜运输荧蒽过程中起关键作用的膜蛋白种类及其作用机制.结果共鉴定到172个差异膜蛋白.通过差异膜蛋白的COG和GO功能分析,发现荧蒽的加入主要影响了红球菌BAP-1细胞膜上与能量、转运相关的膜蛋白.ABC转运蛋白和TonB依赖转运蛋白作为微生物主要的载体蛋白对BAP-1跨膜运输荧蒽起到了重要作用.过氧化氢酶和超氧化物歧化酶在第6d有一个显著上调,作为抗氧化防御机制来保护微生物.各种膜蛋白在不同阶段发挥着各自作用,这些蛋白共同组成蛋白互作网络调控红球菌BAP-1的跨膜运输过程.     

嗜酸硫杆菌双组分信号转导系统多样性分析

为探索嗜酸硫杆菌(Acidithiobacillus)对极端矿山环境条件感知和反应分子机制的差异,利用生物信息学方法鉴定和分析了两株嗜酸氧化亚铁硫杆菌、一株喜温嗜酸硫杆菌和一株耐冷嗜酸硫杆菌的双组分信号转导系统(TCS)结构多样性特征.结果表明,4株嗜酸硫杆菌拥有27~38个TCS蛋白,聚类分析显示有16组配对的TCS族、4组孤儿组氨酸蛋白激酶(HK)和9组孤儿反应调节蛋白(RR),与其进化分支有较好的相关性;其中9个配对和5个不配对的TCS 蛋白族为嗜酸硫杆菌菌株所共有,其余特有TCS均在一个或几个菌株中缺失.嗜酸硫杆菌菌株共有的TCS功能涉及氮素固定及代谢调节、柠檬酸/苹果酸代谢调节等,特有TCS功能涉及如趋化性调控、重金属响应、铜抗性调控等,这可能有助于菌株在极端矿山环境中生存.     

2000—2019年全球猪肉贸易格局演变及其对中国的启示——基于复杂贸易网络分析视角

基于2000—2019年全球猪肉贸易数据,采用复杂网络分析方法,研究了全球猪肉贸易网络格局演变及中国猪肉贸易网络特征,并进一步探讨了中国扩大猪肉进口的潜在贸易伙伴选择问题。研究结果表明：过去近20年来,全球猪肉贸易网络核心国相对固定,猪肉贸易网络呈“碎片化”特征。目前中国已经成为全球第一大猪肉进口国,且进口呈现北美、西欧、南美“三足鼎立”的来源格局。未来中国扩大猪肉进口,潜在的贸易伙伴可选范围比较广泛。一方面可以继续巩固和深化同德国、西班牙、美国、巴西等传统贸易伙伴之间的合作关系,另一方面也可以积极拓展朋友圈,增加从意大利、比利时等其他出口国及泰国、越南等周边国家的进口,以打造更加多元高效的猪肉进口网络。     

'Molecular farming' of antibodies in plants

'Molecular farming' is the production of valuable recombinant proteins in transgenic organisms on an agricultural scale. While plants have long been used as a source of medicinal compounds, molecular farming represents a novel source of molecular medicines, such as plasma proteins, enzymes, growth factors, vaccines and recombinant antibodies, whose medical benefits are understood at a molecular level. Until recently, the broad use of molecular medicines was limited because of the difficulty in producing these proteins outside animals or animal cell culture. The application of molecular biology and plant biotechnology in the 1990s showed that many molecular medicines or vaccines could be synthesised in plants and this technology is termed 'molecular farming'. It results in pharmaceuticals that are safer, easier to produce and less expensive than those produced in animals or microbial culture. An advantage of molecular farming lies in the ability to perform protein production on a massive scale using hectares of cultivated plants. These plants can then be harvested and transported using the agricultural infrastructure. Thus, molecular farming allows rapid progress from genetic engineering to crop production, and new cash crops producing recombinant proteins are already being commercially exploited. We speculate that as functional genomics teaches us more about the nature of disease, molecular farming will produce many of the protein therapeutics that can remedy it.     

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase (APase) exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine (Pinus tabulaeformis), under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, di ered under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the di erent ions played di erent roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.     

生物地质环境与糖尿病的发生

糖尿病是由多种病因引起的糖、蛋白和脂类代谢异常的代谢紊乱性疾病,其发生与发展与生物地质环境密切相关。本文研究了不同地区的糖尿病发病率差异原因以及不同地球化学元素在糖尿病发生发展过程中的作用,认为生物地质环境在糖尿病发生发展过程中起重要作用。根据相同遗传背景的人在不同生物地质环境条件下发病率的差异,提出目前人类糖尿病发病率增加的原因是:片面追求农作物产量,使得谷物、蔬菜和水果中地球化学元素含量降低,进一步引发人类通过食物链从生物地质环境获取地球化学元素绝对量减少以及少量地球化学元素污染生物地质环境引起人体过载。因此,控制人口过快增长,保护环境,实现人类社会与生物地质环境和谐发展,是控制人类糖尿病发病率的关键举措之一。     

Application of proteomics in environmental science

Proteomics involves the separation of proteins, identification of the amino acid sequence of the interested or target proteins, study of the function of the proteins, modification, structure and ultimate assignments to functional pathways in the cell. The proteomic investigations have contributed greatly to human diseases studies, new drugs discovery researches, and environmental science in recent years. This article provides a review on the development of the main proteomic technologies, including both the gel based and non-gel based technologies, and their applications in environmental science. Proteomic technologies have been utilized in the environmental stresses studies to analyze the induction or reduction of proteins at expression level and identify the target proteins to investigate their function in response to environmental stresses, such as high or low pH, oxidation stress, and toxic chemicals. Such protein responses are also helpful to understand the mechanisms of some cellular activities and the functions of some proteins.     

恶臭假单胞菌好氧降解高氯联苯的蛋白质组分析

利用固相pH梯度双向凝胶电泳分离恶臭假单胞菌以葡萄糖和Aroclor1260作为营养基质生长的菌株总蛋白,凝胶经银蓝显色后,采用PDQuest图像分析软件比较、分析识别差异表达蛋白,应用MOLDI-TOF-MS得到相应的肽质量指纹图谱,然后搜索数据库鉴定部分差异蛋白点.结果表明,获得了分辨率较高、重复性较好的不同营养基质条件下菌株蛋白的双向凝胶电泳图谱,比较分析共发现80个差异蛋白点,成功鉴定14个差异表达蛋白,包括应激应答蛋白、蛋白质生物合成、运载体和代谢相关酶类.提示与葡萄糖作为生长基质相比,菌体以多氯联苯作为碳源和能源生长时,产生应激,通过增强应激响应蛋白、蛋白质生物合成相关蛋白和相关代谢酶类的表达使细胞在胁迫条件下维持自身的稳定性与生长代谢.