Crystallin distribution patterns in Litoria infrafrenata and Phyllomedusa sauvagei lenses

The eye lens remains transparent because of soluble lens proteins known as crystallins. For years γ-crystallins have been known as the main lens proteins in lower vertebrates such as fish and amphibians. The unique growth features of the lens render it an ideal structure to study ageing; few studies have examined such changes in anuran lenses. This study aimed to investigate protein distribution patterns in Litoria infrafrenata and Phyllomedusa sauvagei species. Lenses were fractionated into concentric layers by controlled dissolution. Water-soluble proteins were separated into high (HMW), middle (MMW) and low molecular weight (LMW) fractions by size-exclusion HPLC and constituents of each protein class revealed by 1DE and 2DE. Spots were selected from 2DE gels on the basis of known ranges of subunit molecular weights and pH ranges and were identified by MALDI-TOF/TOF MS following trypsin digestion. Comparable lens distribution patterns were found for each species studied. Common crystallins were detected in both species; the most prominent of these was γ-crystallin. Towards the lens centre, there was a decrease in α- and β-crystallin proportions and an increase in γ-crystallins. Subunits representing taxon-specific crystallins demonstrating strong sequence homology with ζ-crystallin/quinone oxidoreductase were found in both L. infrafrenata and P. sauvagei lenses. Further work is needed to determine which amphibians have taxon-specific crystallins, their evolutionary origins, and their function.     

Determination of 19 elements in human eye lenses

Neutron activation analysis was used to determine the concentrations of 19 elements in normal and senile human cataractous lenses. It was found that the concentrations of Ca, Na, Cl, Eu, Sb, and Fe were significantly higher, and those of K, Rb, Cs, Cr, Mn, Co, Sc, and Ce were significantly lower in senile mature cataractous lenses than those in normal human eye lenses. No changes were found for the concentrations of Se, Zn, Mg, S, and Th in the two groups. Positive correlations between Na, Cl, and Ca and K, Rb, and Cs were found, whereas a significantly negative correlation between na, Ca, Cl and K, Rb, Cs were found. The roles of these elements in the evolution of cataract are discussed.     

中华大蟾蜍卵母细胞质膜存在钙依赖性的氯离子通道

本文采用双微电极电压钳方法研究了中华大蟾蜍卵母细胞内源性电压门控型离子通道的成分及其生理特性。卵母细胞去极化至 -30 mV 及更正电压时,有一持续的电压依赖性外向电流出现。钾离子通道拮抗剂四乙基氯化氨(tetraethy-lammonium chloride, TEA, 10 mmol/L)和 4- 氨基吡啶(4-aminopyridine, 4-AP, 10 mmol/L)协同作用时,该电流只能被抑制到最大电流幅度的(23.4±0.72)%。但是,上述浓度的TEA和4-AP 与氯离子通道拮抗剂5- 硝基-2, 3- 苯酚丙胺苯甲酸盐 (5-nitro-2,3-phenypropylamino benzoate, NPPB, 30 μmol/L)、无钙 Ringer 氏液或钙离子通道拮抗剂维拉帕米(40 μmol/L)协同作用时,可分别将此外向电流抑制到最大电流幅度的(2.1±0.08)%、(2.2±0.04)% 和(3.1±0.15)%。结果表明,中华大蟾蜍卵母细胞质膜上除有钾离子电流之外,还存在钙依赖性的氯离子电流。     

Validation of a robust proteomic analysis carried out on formalin‐fixed paraffin‐embedded tissues of the pancreas obtained from mouse and human

A number of reports have recently emerged with focus on extraction of proteins from formalin‐fixed paraffin‐embedded (FFPE) tissues for MS analysis; however, reproducibility and robustness as compared to flash frozen controls is generally overlooked. The goal of this study was to identify and validate a practical and highly robust approach for the proteomics analysis of FFPE tissues. FFPE and matched frozen pancreatic tissues obtained from mice (n = 8) were analyzed using 1D‐nanoLC‐MS(MS)² following work up with commercially available kits. The chosen approach for FFPE tissues was found to be highly comparable to that of frozen. In addition, the total number of unique peptides identified between the two groups was highly similar, with 958 identified for FFPE and 1070 identified for frozen, with protein identifications that corresponded by approximately 80%. This approach was then applied to archived human FFPE pancreatic cancer specimens (n = 11) as compared to uninvolved tissues (n = 8), where 47 potential pancreatic ductal adenocarcinoma markers were identified as significantly increased, of which 28 were previously reported. Further, these proteins share strongly overlapping pathway associations to pancreatic cancer that include estrogen receptor α. Together, these data support the validation of an approach for the proteomic analysis of FFPE tissues that is straightforward and highly robust, which can also be effectively applied toward translational studies of disease.     

云南省三种典型气候带凋落物层弹尾类多样性分布格局

弹尾类是土壤动物中常见的优势类群,其作为土壤微食物网的重要组成部分,参与凋落物分解、土壤团聚体形成等重要生态过程。以往对弹尾类分布格局的研究通常关注其在不同海拔梯度或者不同生境类型下的分布情况,但在不同气候带下弹尾类多样性沿纬度分布格局仍不清楚。为讨论不同气候带下弹尾类沿纬度的分布格局及其潜在的环境影响机制,于2017年10月(雨季末期)在云南省同一经度(E 101°)分布的三种典型气候带设置海拔梯度样带:热带雨林(西双版纳,800 m、1000 m、1200 m、1400 m)、亚热带常绿阔叶林(哀牢山,2000 m、2200 m、2400 m、2600 m)、亚高山针叶林(丽江玉龙雪山,3200 m、3400 m、3600 m、3800 m),采集凋落物层弹尾类并调查分析土壤温度、土壤含水量、凋落物厚度、土壤pH、土壤容重及土壤孔隙度等环境因子。利用Berlese-Tullgren法收集土壤动物,共获得弹尾类19150只,隶属于10科29属,其中符■属(39.9%,等节■科)数量最多,其余优势属为棘■属(21.7%,棘■科)和球角■属(10.1%,球角■科),这3个属合计占总体的71.7%。凋落物层弹尾类的密度在亚高山针叶林明显高于热带雨林和亚热带常绿阔叶林,弹尾类的属数排序由多到少依次为热带雨林、亚高山针叶林、亚热带常绿阔叶林。通过最小二乘回归法对弹尾类多样性的海拔格局进行回归分析,得出弹尾类的丰富度指数(Margalef′s指数、Menhinick′s指数)、多样性指数(Shannon-Wiener多样性指数、Simpson多样性指数)和均匀度指数(Pielou均匀度指数)沿热带雨林、亚热带常绿阔叶林、亚高山针叶林呈单调递减格局。通过全模型子集回归筛选最佳环境模型表明,温度是影响弹尾类多样性沿不同气候带分布格局的主要环境因子。本研究为预测不同气候带下弹尾类多样性如何响应环境变化提供参考。     

Quantitative evaluation of tryptophan oxidation in actin and troponin I from skeletal muscles using a rat model of acute oxidative stress

Increased levels of “ROS” cause oxidative stress and are believed to play a key role in the development of age‐related diseases and mammalian aging, e.g. through the oxidation of residues, at or close to, the protein surface. In this study, we have investigated the effects of ROS on tryptophan residues in alpha skeletal actin and troponin I (fast skeletal muscle isoform) using an established rat model of acute oxidative stress induced by X‐ray irradiation. In the control samples (no oxidative stress), the single Trp residue of troponin I (position 161) and the four tryptophan residues present in actin (positions 79, 86, 340, and 356) were only oxidized at very low levels. Post‐irradiation, the level of oxidized versions increased for most positions within 3 h. Tryptophan residues located inside the proteins, however, required longer time periods. Based on the increment masses of the tryptophan positions calculated from the b‐ and y‐ion series of the tandem mass spectra, the following oxidation products of tryptophan were detected: kynurenine; oxolactone; hydroxytryptophan or oxindolylalanine (isobaric); hydroxykynurenine; dioxindolylalanine, N‐formylkynurenine or dihydroxytryptophan (all three isobaric); and hydroxyl‐N‐formylkynurenine, with mass gains relative to tryptophan of 4, 14, 16, 20, 32, and 48 u, respectively. Despite a partial recovery after 24 h, the degree of oxidation of all oxidized versions was still higher than in the control samples.     

Evaluation of protein expression in bovine bronchoalveolar fluid following challenge with Mannheimia haemolytica

Proteomics analysis of bovine bronchoalveolar fluid (BAF) following induction of pneumonia with Mannheimia haemolytica using nanoflow liquid chromatography coupled with tandem mass spectrometry (nanoLC-MS/MS) resulted in the identification of 88 unique proteins. Proteins detected in BAF included antimicrobial peptides (AMPs), complement factors, acute-phase proteins, protease inhibitors, and proteins involved in oxidation-reduction. Notwithstanding biological variation, differences in relative protein abundance, determined using normalized peptide counts, were detected for select proteins in BAF from genuinely infected versus sham-infected animals. To demonstrate the applicability of using normalized peptide counts to assess protein expression trends, LC-MS/MS data for the acute-phase protein haptoglobin (HPT) were compared with ELISA data, and statistical evaluation of the relationship between the data revealed a strong measure of association. Differences were detected between sham- and genuinely infected animals for haptoglobin, as well as the AMPs cathelicidin-1 and cathelicidin-4, and inter-α-trypsin inhibitor heavy chain-4, a fairly novel protein involved in the acute phase response. Though the small sample size limited the scope of the inferences, the results indicate the likely importance of AMPs and acute-phase proteins during respiratory infection, and provide additional information regarding potential mechanisms involved in the bovine mucosal barrier defense.     

A multicenter study of radiation doses to the eye lenses of medical staff performing non-vascular imaging and interventional radiology procedures in Japan

PurposeThis study aimed to measure the eye lens doses received by physicians and other medical staff participating in non-vascular imaging and interventional radiology procedures in Japan.Material and methodsFrom October 2014 to March 2017, 34 physicians and 29 other medical staff engaged in non-vascular imaging and interventional radiology procedures at 18 Japanese medical facilities. These professionals wore radioprotective lead glasses equipped with small, optically stimulated luminescence dosimeters and additional personal dosimeters at the neck during a 1-month monitoring period. The H_p(3) and the H_p(10) and H_p(0.07) were obtained from these devices, respectively. The monthly H_p(3), H_p(10), and H_p(0.07) for each physician and other medical staff member were then rescaled to a 12-month period to enable comparisons with the revised occupational equivalent dose limit for the eye lens.ResultsAmong physicians, the average annual H_p(3) values measured by the small luminescence dosimeters on radioprotective glasses were 25.5 ± 38.3 mSv/y (range: 0.4–166.8 mSv/y) and 9.3 ± 16.6 mSv/y (range: 0.3–82.4 mSv/y) on the left and right sides, respectively. The corresponding values for other medical staff were 3.7 ± 3.1 mSv/y (range: 0.4–10.4 mSv/y) and 3.2 ± 2.7 mSv/y (range: 0.5–11.5 mSv/y), respectively.ConclusionsThe eye lens doses incurred by physicians and other medical staff who engaged in non-vascular imaging and interventional radiology procedures in Japan were provided. Physicians should wear radioprotective glasses and use additional radioprotective devices to reduce the amount of eye lens doses they receive.     

Identification of 'Campylobacter upsaliensis' and other catalase-negative campylobacters from paediatric blood cultures by numerical analysis of electrophoretic protein patterns

Twenty-one strains of catalase-negative campylobacters from paediatric blood cultures were characterized by one-dimensional SDS-PAGE of cellular proteins. A further 11 Campylobacter strains were included for reference purposes. The partial protein patterns were used as the basis of a numerical analysis, which showed that 17 hippurate-negative strains had a high similarity to 'C. upsaliensis' (r greater than or equal to 0.82) irrespective of their geographical location, and that three hippurate-positive isolates had a high similarity (r greater than or equal to 0.87) to C. jejuni subsp. jejuni. One hippurate-positive CNW strain was not identified. The analysis of SDS-PAGE protein patterns proved an excellent method of characterizing these thermophilic campylobacter as they were difficult to identify by traditional methods.     

Combining selected reaction monitoring with discovery proteomics in limited biological samples

Simultaneous quantification of multiple proteins by selected reaction monitoring (SRM) has several applications in cell signaling studies including embryo proteomics. However, concerns have recently been raised over the specificity of SRM assays due to possible ion redundancy and/or sequence similarity of selected peptide with multiple non‐related proteins. In this Viewpoint article, we discuss some simple measures that can increase our confidence in the accuracy of SRM scans used in proteomic experiments. At least in embryonic samples from porcine species, these measures were found to be useful in validating MS‐identified differentially expressed proteins. Among the nine proteins analyzed by SRM assay, all the proteins that were found to be up‐ or down‐regulated in MS experiment were also faithfully up‐ or down‐regulated in SRM assay.     

Geographical patterns of genetic variability in introduced Australian populations of the marine toad,Bufo marinus: Sorbitol dehydrogenase,Sdh

The marine toad, Bufo marinus, was introduced to Australia from Hawaii in 1935. From 1935 to 1974, the toad population expanded exponentially to occupy 584,000 km², and now has a continuous distribution from Cape York to Tweed River on the eastern coast of the continent. Genetic analysis of the population indicates a difference in allele frequency at the sorbitol dehydrogenase locus. There are two alleles segregating at the locus (NAD-Sdh^a and NAD-Sdh^b). The NAD-Sdh^aa homozygote is common in the two southern populations, but uncommon in northern populations. The north-south difference has been established in less than 25 generations.     

Proteomics of the milk fat globule membrane from Camelus dromedarius

Camel milk has been widely characterized with regards to casein and whey proteins. However, in camelids, almost nothing is known about the milk fat globule membrane (MFGM), the membrane surrounding fat globules in milk. The purpose of this study was thus to identify MFGM proteins from Camelus dromedarius milk. Major MFGM proteins (namely, fatty acid synthase, xanthine oxidase, butyrophilin, lactadherin, and adipophilin) already evidenced in cow milk were identified in camel milk using MS. In addition, a 1D‐LC‐MS/MS approach led us to identify 322 functional groups of proteins associated with the camel MFGM. Dromedary MFGM proteins were then classified into functional categories using DAVID (the Database for Annotation, Visualization, and Integrated Discovery) bioinformatics resources. More than 50% of MFGM proteins from camel milk were found to be integral membrane proteins (mostly belonging to the plasma membrane), or proteins associated to the membrane. Enriched GO terms associated with MFGM proteins from camel milk were protein transport (p‐value = 1.73 × 10^?14), translation (p‐value = 1.08 × 10^?11), lipid biosynthetic process (p‐value = 6.72 × 10^?10), hexose metabolic process (p‐value = 1.89 × 10^?04), and actin cytoskeleton organization (p‐value = 2.72 × 10^?04). These findings will help to contribute to a better characterization of camel milk. Identified MFGM proteins from camel milk may also provide new insight into lipid droplet formation in the mammary epithelial cell.     

A novel type of crystallin in the frog eye lens. 35-kDa polypeptide is not homologous to any of the major classes of lens crystallins

The nucleotide sequence of a cloned DNA coding for the 35-kDa polypeptide of the eye lens of the frog (Rana temporaria) has been determined. The sequence without connectors and poly(A) tract is 889 nucleotides in length and shows no homology with sequences coding for other classes of crystallins: alpha-, beta-, gamma- or delta-crystallins. The sequence contains one reading frame 675 nucleotides in length, an apparently intact 3'-non-translated region with the polyadenylation signal sequence and a poly(A) tract; the 5'-non-translated region is lost along with part of the coding region; this accounts for about 1/4 of the total mRNA length. The secondary structure prediction according to the Ptitsin - Finkelstein method shows the presence of predominantly beta-strands with only a few alpha-helical regions. We conclude that the 35-kDa polypeptide from the frog eye lens belongs to a new class of eye lens crystallins for which we propose the name epsilon-crystallin.     

Invasive alien plant species in China: regional distribution patterns

Plant invasions have been attracting increasing attention from ecologists because of their worldwide environmental impacts and huge economic costs. Research on the characteristics of the recipient regions is essential for understanding the process of plant invasion. However, few previous studies on invasibility of habitats include social factors, although human activities are critical in the process of plant invasion. China is a vast country with high plant species diversity and a long history of introduction of exotic plant species and is particularly vulnerable to invasive plant species. Alien plant species are widespread in the country. Therefore, the study of invasive plants in China is urgent in practice and theoretically important for developing invasion ecology. For the present study, 126 species were selected to represent the major invasive plant species in China. We then collected data on their species richness in 31 provincial administrative units of China and performed Spearman rank correlations between species richness and possible natural and socio‐economic factors. We found that socio‐economic factors, such as human density and GDP, correlated positively with the species richness of invasive plants in China. In conjunction with the natural and socio‐economic correlations in the study of regional distribution pattern of the major invasive plants, we discussed the factors influencing the regional distribution pattern of the major invasive plants in China. We suggest that native plant species richness was mainly determined by the natural conditions of the regions, while invasive species richness was influenced by natural conditions and human disturbance together.     

The absence of the long 3'-non-translated region in mRNA coding for eye lens alpha A2-crystallin of the frog (Rana temporaria)

The nucleotide sequence of a cloned cDNA (clone pRt(1)297; GENE (1982) 17, 131) coding for a 18 kDa polypeptide of the frog eye lens has been determined. The sequence, 791 nucleotide in length has only one long open reading frame (447 nucleotides). The derived amino acid sequence in this frame has greater than 90% homology with the region 25-173 of alpha A2-crystallin amino acid sequence from a related frog species Rana pipiens. The 5'-terminal part of mRNA corresponding to the first 24 amino acids of alpha A2-crystallin has been lost in cloning and substituted by an artefactual sequence. The 3'-terminal part appears to be intact as follows from the presence of the universal poly(A) addition site and poly(A) tract. The 3'-nontranslated region present in frog alpha A2-crystallin mRNA (130 nucleotides) is about 4-times shorter than in mammalian alpha A2-crystallin mRNA. Intact alpha A2-crystallin mRNA with a size of about 700 nucleotides as determined by Northern blot hybridization is about twice smaller than corresponding mammalian mRNAs.     

Scots pine root distribution derived from radial sap flow patterns in stems of large leaning trees

This study characterizes whole tree root system distribution in a non-destructive way based on its functional parameters, particularly the sap flow patterns in stems. This approach particularly considers sap flow variation across stems, both radial and circumferential patterns of flow that are usually used for a better integration of sap flow density at the whole tree level. We focused at: (1) Showing examples of sap flow variation across stems at a defined situation (high midday values at the period of non-limiting water supply; (2) Analyzing radial flow patterns in terms of root distribution; (3) Validating these results at the stand level (mean data of series of individual trees) using results of classical biometric methods used at the same site; and (4) Applying the results for evaluation of root distribution around leaning trees. Sap flow rate was measured by the heat deformation method on a set of 14 trees at an experimental pine forest stand in Brasschaat (Belgium) during the growing season of 2000. Sap flow variation across stems was measured at a total of 700 points. Amounts of water supplied by superficial (horizontally oriented) and sinker (vertically oriented) roots were estimated from sap flow patterns. The vertical distribution of absorbing roots as derived from the analysis of sap flow patterns in stem sapwood was very similar to the distribution determined by the classical biometric analysis of fine roots. Trees leaning to the East had stem radii at the stump level and crown radii enhanced in the leaning direction. Sinker roots showed higher absorption activities in the leaning direction, but superficial roots were more absorbing in the opposite direction. The application of the above-described method allows for a better evaluation of the whole-tree behavior and facilitates the evaluation of tree and stand properties in traditional forest stands, which are not equipped for detailed scientific research. This may also facilitate practical applications in landscape-level studies.     

Proteomic analysis of physiological function response to hot summer in liver from lactating dairy cows

Lactation performance of dairy cattle is susceptible to heat stress. The liver is one of the most crucial organs affected by high temperature in dairy cows. However, the physiological adaption by the liver to hot summer conditions has not been well elucidated in lactating dairy cows. In the present study, proteomic analysis of the liver in dairy cows in spring and hot summer was performed using a label-free method. In total, 127 differentially expressed proteins were identified; most of the upregulated proteins were involved in protein metabolic processes and responses to stimuli, whereas most of the downregulated proteins were related to oxidation–reduction. Pathway analysis indicated that 3 upregulated heat stress proteins (HSP90α, HSP90β, and endoplasmin) were enriched in the NOD-like receptor signaling pathway, whereas several downregulated NADH dehydrogenase proteins were involved in the oxidative phosphorylation pathway. The protein–protein interaction network indicated that several upregulated HSPs (HSP90α, HSP90β, and GRP78) were involved in more interactions than other proteins and were thus considered as central hub nodes. Our findings provide novel insights into the physiological adaption of liver function in lactating dairy cows to natural high temperature.     

陷阱法调查棉田蜘蛛的分布动态

采用陷阱法系统调查棉田各发育期的蜘蛛群落分布型,运用5种聚集度指标、Taylor的幂法则以及m*--x的回归关系综合分析。结果显示:(1)陷阱法所采集的蜘蛛绝大多数为游猎型蜘蛛。(2)棉田游猎性蜘蛛群落在整个棉花发育期都呈随机分布。