Development and validation of stable reference materials for food microbiology using Bacillus cereus and Clostridium perfringens spores

Aims: To develop a new type of microbiological Reference Materials (RMs), displaying long‐term stability at room temperature. The purpose was to produce and validate two batches of RMs for the enumeration of Bacillus cereus and Clostridium perfringens. Methods and Results: The RMs were based on spores of B. cereus and Cl. perfringens, adsorbed on calcium carbonate pellets. Two batches of 1000 units were manufactured and validated in compliance with ISO guide 35. After verification of their homogeneity, the stability of the ‘RM‐B. cereus’ and ‘RM‐Cl. perfringens’ batches was proven during at least 36 and 9 months, respectively, at room temperature. The validation study was completed by international collaborative trial involving 12 laboratories, allowing the validation of the assigned values. Conclusions: The methodology developed in this work enabled to produce easy‐to‐handle and cost‐effective RMs, displaying an unprecedented stability at room temperature, a good homogeneity and a precise and validated assigned value. Significance and Impact of the Study: This study revealed new paths for the development of stable microbiological RMs. Overcoming the intrinsic instability of the living cells makes it possible to produce valuable tools for the quality assurance of microbiology laboratories.     

组织工程面临的技术挑战与发展趋势

当前组织工程研究仍处于初级阶段,还仅仅是初步应用组织工程技术修复临床简单组织缺损,还有许多制约组织工程应用与发展的基本科学问题没有阐明.随着组织工程各个层面技术难题的逐个攻破,组织工程的内涵和外延将不断拓展,并有助于加快组织工程的产业化进程,促进临床应用.针对组织工程核心要素研究的不足,结合最新的组织工程研究进展,阐述...     

Hemicellulosic polymers of cabbage leaves

The hemicellulosic polymers of depectinated cell-wall material of immature cabbage leaves have been extracted by alkali, fractionated by ion-exchange chromatography and their structural features studied. In the 1 M potassium hydroxide-soluble fraction the main polymers are arabinoxylan-xyloglucan-pectic, arabinoxylan-pectic-protein and arabinoxylan-xyloglucan-pectic-protein complexes. Small amounts of polysaccharide-protein-polyphenol complexes are also present. In the 4 M potassium hydroxide-soluble fraction the predominant polymers are two xyloglucans, the major one of which appears to have ca 10% of(1 → 4)-linked and 3% of(1 → 4,6)-linked mannose residues associated with it, and both have terminal galactose, fucose and possibly arabinose residues in the side chains. Methylation analysis of the oligosaccharides, formed by degradation with cellulase, and partial hydrolysis of the methylated xyloglucan, followed by re-methylation with CD₃I, have enabled the formulation of a tentative structure which is similar to other plant galactoxyloglucans. It was not possible to establish whether the mannose residues are part of an associated glucomannan or whether they are an integral part of the glucan backbone. The general structural features of the hemicellulosic polymers are discussed in the light of these results.     

A Study of Glutathione <Emphasis Type="Italic">S</Emphasis>-transferase pi Expression in Central Nervous System of Subjects with Amyotrophic Lateral Sclerosis Using RNA Extraction from Formalin-Fixed,Paraffin-Embedded Material

The expression of glutathione S-transferase pi (GST pi), an enzyme responsible for inactivation of a large variety of toxic compounds was studied in spinal cord, motor and sensory brain cortex obtained from patients who died in the course of amyotrophic lateral sclerosis (ALS). The studies were performed on formalin-fixed, paraffin-embedded (FFPE) and freshly frozen tissues. The method of RNA isolation from FFPE was modified. A significant decrease of GST pi-mRNA expression was found in cervical spinal cord and motor brain cortex of ALS subjects comparing to analogue control tissues (P < 0.01), as well as in motor cortex of ALS subjects comparing to their sensory cortex (P < 0.05). In spinal cords the decrease in GST pi-mRNA expression was accompanied by a decrease of GST pi protein level. Results indicated lowered GST pi expression on both mRNA and protein levels in the regions of nervous system affected by ALS. The non-properly inactivated by GST toxic electrophiles and organic peroxides may thus contribute to motor neurons damage.     

Effects of ambient air particles on the endothelin system in human pulmonary epithelial cells (A549)

Inhalation of urban particles results in higher circulating levels of the vasoconstrictor peptide endothelin-1 (ET-1), which may account for the adverse cardiovascular impacts associated with air pollution. The objective of this study was to examine the direct effects of urban particles on the production of ET-1 by human epithelial cells (A549). A549 cells were exposed to TiO₂, SiO₂, Ottawa urban particulate matter EHC-93, and fractions of the urban particles. The levels of ET-1, interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF) in the culture medium were detected by ELISA. The mRNA levels of preproET-1, endothelin converting enzyme (ECE-1), ETa receptor and ETb receptor, matrix metalloproteinase (MMP-2), tissue inhibitor of MMP (TIMP-2), and heat shock protein (HSP-70) were determined by quantitative real-time RT-PCR. Cluster analysis of the variables identified similarities in the patterns of effects. Cluster I comprised variables that were primarily inhibited by particles: ET-1 and MMP-2 mRNAs, ET-1 and bigET-1 peptides, and cell viability. Clusters II and III comprised variables that were either inhibited or induced, depending on the test material: HSP-70, ETaR and ECE mRNAs, and IL-8 and VEGF proteins. Cluster IV comprised variables that were mainly induced by particle preparations: ETbR and TIMP-2 mRNAs. The decreased expression of preproET-1 in A549 cells suggests that epithelial cells may not be the source of higher pulmonary ET-1 spillover in the circulation measured in vivo in response to inhaled urban particles. However, higher ECE-1 in A549 cells after exposure to particles suggests an increased ability to process bigET-1 into the mature ET-1 peptide, while increased receptor expression implies higher responsiveness. The increased release of IL-8 and VEGF by epithelial cells in response to particles could possibly upregulate ET-1 production in the adjacent pulmonary capillary endothelial cells, with concomitant increased ET-1 spillover in the systemic circulation.     

Lipophilic flavones of Primula veris L. from field cultivation and in vitro cultures

Ten lipophilic flavones were isolated from the leaves of Primula veris from field cultivation - the newly described 3'-hydroxy-4',5'-dimethoxyflavone and 3'-methoxy-4',5'-methylenedioxyflavone, the previously known from chemical synthesis 3',4'-dimethoxyflavone, 2',5'-dimethoxyflavone, and also flavone, 2'-hydroxyflavone, 2'-methoxyflavone, 3'-methoxyflavone, 3',4',5'-trimethoxyflavone and 5,6,2',6'-tetramethoxyflavone (zapotin) which were previously known from plants. The same flavones were found in the leaves of P. veris obtained by in vitro propagation. The structural assignments were derived from (1)H NMR, (13)C NMR, EIMS and UV spectral data and the influence of B-ring oxygen substituents on the C-2, C-3 and H-3 NMR resonances in flavones unsubstituted in the A ring is taken into consideration.     

Waste Treatment and Assessment of Long-Term Emissions (8pp)

Goal, Scope and Background The disposal phase of a product’s life cycle in LCA is often neglected or based on coarse indicators like ‘kilogram waste’. The goal of report No. 13 of the ecoinvent project (Doka 2003) is to create detailed Life Cycle Inventories of waste disposal processes. The purpose of this paper is to give an overview of the models behind the waste disposal inventories in ecoinvent, to present exemplary results and to discuss the assessment of long-term emissions. This paper does not present a particular LCA study. Inventories are compiled for many different materials and various disposal technologies. Considered disposal technologies are municipal incineration and different landfill types, including sanitary landfills, hazardous waste incineration, waste deposits in deep salt mines, surface spreading of sludges, municipal wastewater treatment, and building dismantling. The inventoried technologies are largely based on Swiss plants. Inventories can be used for assessment of the disposal of common, generic waste materials like paper, plastics, packaging etc. Inventories are also used within the ecoinvent database itself to inventory the disposal of specific wastes generated during the production phase. Inventories relate as far as possible to the specific chemical composition of the waste material (waste-specific burdens). Certain expenditures are not related to the waste composition and are inventoried with average values (process-specific burdens). Methods The disposal models are based on previous work, partly used in earlier versions of ecoinvent/ETH LCI data. Important improvements were the extension of the number of considered chemical elements to 41 throughout all disposal models and new landfill models based on field data. New inventories are compiled for waste deposits in deep salt mines and building material disposal. Along with the ecoinvent data and the reports, also Excel-based software tools were created, which allow ecoinvent members to calculate waste disposal inventories from arbitrary waste compositions. The modelling of long-term emissions from landfills is a crucial part in any waste disposal process. In ecoinvent long-term emissions are defined as emissions occurring 100 years after present. They are reported in separate emission categories. The landfill inventories include long-term emissions with a time horizon of 60’000 years after present. Results and Discussion As in earlier studies, the landfills prove to be generally relevant disposal processes, as also incineration and wastewater treatment processes produce landfilled wastes. Heavy metals tend to concentrate in landfills and are washed out to a varying degree over time. Long-term emissions usually represent an important burden from landfills. Comparisons between burdens from production of materials and the burdens from their disposal show that disposal has a certain relevance. Conclusion The disposal phase should by default be included in LCA studies. The use of a material not only necessitates its production, but also requires its disposal. The created inventories and user tools facilitate heeding the disposal phase with a similar level of detail as production processes. The risk of LCA-based decisions shifting burdens from the production or use phase to the disposal phase because of data gaps can therefore be diminished. Recommendation and Perspective Future improvements should include the modelling of metal ore refining waste (tailings) which is currently neglected in ecoinvent, but is likely to be relevant for metals production. The disposal technologies considered here are those of developed Western countries. Disposal in other parts of the World can differ distinctly, for logistic, climatic and economic reasons. The cross-examination of landfill models to LCIA soil fate models could be advantageous. Currently only chemical elements, like copper, zinc, nitrogen etc. are heeded by the disposal models. A possible extension could be the modelling of the behaviour of chemical compounds, like dioxins or other hydrocarbons.     

The restoration of anatomic form of atrophied alveolar processes of jaws with the help of bioimplant lioplast

The purpose of the present research was: 1. To investigate the opportunity of application of allogen osteoplastic materials such as Lioplast, received in Samara Tissue Bank of Samara State Medical University. 2. To work out a new technique of producing lyophilized allogen osteoplastic materials, such as Lioplast, with application of an ultrasonic method of clearing. 3. The development of various techniques of operative intervention on jaws for increase in volume of atrophied, as a result of secondary adentia, alveolar shoot. 4. The application of spongy allogen osteoplastic material as a skeleton for filling in surgical operations on jaws. 5. The demonstration of the Technique of a bone-periostic Lioplast 'sandwich' for formation of an alveolar shoot.     

A taxonomic review of the nivicolous species of myxomycetes described by Kowalski. I. Order Stemonitales

Five type specimens of species from the order Stemonitales proposed by Kowalski are re-examined. These are Comatricha fusiforme, Diacheopsis effusa, D. serpula, Lamproderma acanthosporum and L. disseminatum. SEM and LM micrographs of spores, capillitia and sporocarps are provided. Critical study of these type specimens supports Kowalski’s recognition of these taxa as good and valid species.     

沙漠石蜂的筑巢材料和蜂巢结构

沙漠石蜂Megachile(Chalicodoma) desertorum Morawitz为多种植物的传粉昆虫,在西鄂尔多斯地区是多种濒危植物和荒漠建群植物的重要传粉者。在该地区研究沙漠石蜂的筑巢场所、筑巢材料和蜂巢结构。共调查统计5148个蜂巢的筑巢场所和朝向;选择4个土壤表层成分不同的筑巢地,分别测定其筑巢沙土的机械组成、pH值和土壤紧实度;定点连续观察整个筑巢过程。结果表明:沙漠石蜂95.80%的巢建于岩石表面;土墙、灌木枝条、水泥柱及钢窗框上的巢室数均较少。83.1%的巢室建在东向一面;南向14.2%,西向2.4%,北向仅0.3%;筑巢材料为沙土和唾液制成的泥浆,所用沙土的pH值在8.18～9.34之间、紧实度在1.28～1.84g/cm3之间,其中粒径为0.06～0.15mm的沙土比例最高;1个蜂巢一般由4～10个巢室组成。说明该蜂喜欢将巢室建在朝向东方的岩石表面;筑巢材料主要由唾液和以碱性的粒径为0.06～0.15mm左右的沙土构成。也说明是对干旱炎热的荒漠环境的适应。