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Tested Samples (tested + sample)
Selected AbstractsWhat determines the degree of compactness of a calcium-binding protein?FEBS JOURNAL, Issue 4 2009Liliane Mouawad The EF-hand calcium-binding proteins may exist either in an extended or a compact conformation. This conformation is sometimes correlated with the function of the calcium-binding protein. For those proteins whose structure and function are known, calcium sensors are usually extended and calcium buffers compact; hence, there is interest in predicting the form of the protein starting from its sequence. In the present study, we used two different procedures: one that already exists in the literature, the sosuidumbbell algorithm, mainly based on the charges of the two EF-hand domains, and the other comprising a novel procedure that is based on linker average hydrophilicity. The linker consists of the residues that connect the domains. The two procedures were tested on 17 known-structure calcium-binding proteins and then applied to 59 unknown-structure centrins. The sosuidumbbell algorithm yielded the correct conformations for only 15 of the known-structure proteins and predicted that all centrins should be in a closed form. The linker average hydrophilicity procedure discriminated well between all the extended and non-extended forms of the known-structure calcium-binding proteins, and its prediction concerning centrins reflected well their phylogenetic classification. The linker average hydrophilicity criterion is a simple and powerful means to discriminate between extended and non-extended forms of calcium-binding proteins. What is remarkable is that only a few residues that constitute the linker (between 2 and 20 in our tested sample of proteins) are responsible for the form of the calcium-binding protein, showing that this form is mainly governed by short-range interactions. [source] Rubber tree (Hevea brasiliensis) trunk phloem necrosis: aetiological investigations failed to confirm any biotic causal agentFOREST PATHOLOGY, Issue 1 2007F. Pellegrin Summary Trunk phloem necrosis (TPN) is currently a main constraint in rubber (Hevea brasiliensis) plantations. The apparent spread of the disease, from tree to tree along the planting line, strongly supported the implication of a pathogen that could be transmitted mechanically via the tapping knife. In order to detect a causal agent of the disease, studies focusing on characterization of the known mechanically transmitted pathogens (e.g. viroids, cryptic viruses or phytoplasma) were initiated. RNA strands of low molecular weight (200,400 and >500 bp) displaying structural similarities with viroids and viral dsRNAs were observed in various tested samples. However, attempts to show the potential role of these RNA molecules in the spread of the disease failed. First of all, there was no significant or reproducible correlation between the health status of the rubber trees sampled and these RNA molecules. Moreover, no sequence homology with known pathogens could be found when randomly amplified cDNA fragments isolated from trees presenting the disease symptoms were sequenced. In conclusion, the aetiological investigations, in order to show the presence of a pathogen responsible of the TPN disease, were non-conclusive, which tends to disprove the hypothesis of a biotic causal agent. [source] Ace2, rather than ace1, is the major acetylcholinesterase in the silkworm, Bombyx moriINSECT SCIENCE, Issue 4 2009Hui-Juan Chen Abstract, Two acetylcholinesterase (ace) genes have been reported in many insect species. In pests such as Helicoverpa assulta and Plutella xylostellas, ace1 gene encodes the predominant synaptic enzyme that is the main target of organophosphorus (OP) and carbamate pesticides. It has been reported that pesticide selection has an impact on the ace gene evolution. The domesticated silkworm, Bombyx mori, also has two ace genes. We studied ace gene expression and enzyme activities in silkworm as this has not faced pesticide selection over the past decades. The expression levels of two ace genes, Bm- ace1 and Bm- ace2, were estimated by quantitative real-time polymerase chain reaction. Bm- ace2 was expressed more highly than Bm- ace1 in all tested samples of different developmental stages or tissues, suggesting ace2, rather than ace1, is the major type of acetylcholinesterase (AChE) in Bombyx mori. This is inconsistent with the aforementioned lepidopterons agricultural pests, partly be due to the widespread use of pesticides that may induce high expression of the ace1 gene in these pests. Besides high expression in the head, Bm- ace1 also expresses highly in the silk glands and Bm- ace2 is abundant in the germline, implying both ace genes may have potential non-hydrolytic roles in development. Furthermore, we found that the mRNA levels of two ace genes and their ratios (ace2/ace1) change day to day in the first and third instars. This challenges the conventional method of estimating enzymatic activity using crude extract as an enzyme solution, as it is a mixture of AChE1 and AChE2. An efficient and simple method for separating different AChEs is necessary for reliable toxicological analyses. [source] Effectiveness of ozone against endodontopathogenic microorganisms in a root canal biofilm modelINTERNATIONAL ENDODONTIC JOURNAL, Issue 1 2009K. C. Huth Abstract Aim, To assess the antimicrobial efficacy of aqueous (1.25,20 ,g mL,1) and gaseous ozone (1,53 g m,3) as an alternative antiseptic against endodontic pathogens in suspension and a biofilm model. Methodology,Enterococcus faecalis, Candida albicans, Peptostreptococcus micros and Pseudomonas aeruginosa were grown in planctonic culture or in mono-species biofilms in root canals for 3 weeks. Cultures were exposed to ozone, sodium hypochlorite (NaOCl; 5.25%, 2.25%), chlorhexidine digluconate (CHX; 2%), hydrogen peroxide (H2O2; 3%) and phosphate buffered saline (control) for 1 min and the remaining colony forming units counted. Ozone gas was applied to the biofilms in two experimental settings, resembling canal areas either difficult (setting 1) or easy (setting 2) to reach. Time-course experiments up to 10 min were included. To compare the tested samples, data were analysed by one-way anova. Results, Concentrations of gaseous ozone down to 1 g m,3 almost and aqueous ozone down to 5 ,g mL,1 completely eliminated the suspended microorganisms as did NaOCl and CHX. Hydrogen peroxide and lower aqueous ozone concentrations were less effective. Aqueous and gaseous ozone were dose- and strain-dependently effective against the biofilm microorganisms. Total elimination was achieved by high-concentrated ozone gas (setting 2) and by NaOCl after 1 min or a lower gas concentration (4 g m,3) after at least 2.5 min. High-concentrated aqueous ozone (20 ,g mL,1) and CHX almost completely eliminated the biofilm cells, whilst H2O2 was less effective. Conclusion, High-concentrated gaseous and aqueous ozone was dose-, strain- and time-dependently effective against the tested microorganisms in suspension and the biofilm test model. [source] Rapid assessment of the sex of codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) eggs and larvaeJOURNAL OF APPLIED ENTOMOLOGY, Issue 4 2009I. Fuková Abstract Two different methods were tested to identify the sex of the early developmental stages of the codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) with a WZ/ZZ (female/male) sex chromosome system. First, it was shown that the sex of all larval stages can be easily determined by the presence or absence of sex chromatin, which is formed by the female-specific W chromosome in interphase nuclei. This trait can also be used to identify the sex of newly hatched larvae but it does require care and accuracy. Secondly, a new sexing technique was developed based on a molecular marker of the codling moth W chromosome. Flanking regions of an earlier described W-specific sequence (CpW2) were isolated and sequenced and a 2.74 kb sequence (CpW2- EcoRI), specific for the W chromosome, was obtained. Several PCR tests were conducted, which confirmed that the CpW2- EcoRI sequence is a reliable marker for the sex identification in codling moth samples of different geographical origin. In addition, a fragment of a codling moth gene, period (Cpper) was isolated and sequenced. Results of southern hybridization of the Cpper probe with female and male genomic DNA suggested that the Cpper gene is located on the Z chromosome. Then a multiplex PCR assay was developed, which co-amplified the CpW2- EcoRI sequence to identify the W chromosome and the Z-linked Cpper sequence, which served as a positive control of accurate processing of tested samples. The multiplex PCR provides an easy and rapid identification of the sex of embryos and early larval instars of the codling moth. [source] Can the high levels of human verocytotoxigenic Escherichia coli O157 infection in rural areas of NE Scotland be explained by consumption of contaminated meat?JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2007O. Solecki Abstract Aims:, To determine if contamination levels of Escherichia coli O157 and generic E. coli in retail-minced meat products are greater in rural shops compared with urban shops in Grampian, NE Scotland. We also investigated whether meat from supermarkets and meat from local butcher shops had a similar bacteriological quality. Methods and Results:, Minced beef and minced lamb were tested from November 2004 to August 2006. Escheichia coli O157 was found at low levels in four samples out of 530 tested samples (0·75%). Generic E. coli were present in 11% of the samples tested, of which 67% came from supermarkets. We observed no significant difference in the prevalence of generic E. coli between rural and urban areas. Conclusions:, Low levels of contamination with E. coli O157 and generic E. coli in retail meat suggest that meat is not a major route of infection in NE Scotland. Significance and Impact of the Study:, The study does not suggest that the high incidence of E. coli O157 human infection in the rural areas of Grampian is because of meat consumption , this provides further evidence of contact with animals or water being the routes of infection. Hence, risk mitigation should be focussed more on environmental pathways of infection. [source] Rietveld structure and in vitro analysis on the influence of magnesium in biphasic (hydroxyapatite and ,-tricalcium phosphate) mixturesJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2009S. Kannan Abstract The structure of two different Mg-substituted biphasic (HAP and ,-TCP) mixtures along with the biphasic mixtures without substituted Mg2+ was investigated using Rietveld refinement technique. The substituted Mg2+ was found in the ,-TCP phase and its influence on the composition has led to an increase in HAP content of Mg-containing biphasic mixtures when compared with the HAP content detected in pure biphasic mixtures. The refined structural parameters of Ca10(PO4)6(OH)2 and ,-Ca3(PO4)2 confirmed that all the investigated compositions have crystallized in the corresponding hexagonal (space group P63/m) and rhombohedral (space group R3c) structures. The substitution of lower sized magnesium was found preferentially incorporated at the sixfold-coordinated Ca (5) site of ,-TCP, which is due to the strong Ca (5)·O interaction among all the five different Ca sites of ,-Ca3(PO4)2. The in vitro tests using primary culture of osteoblasts showed that all the tested samples are biocompatible and promising materials for in vivo studies. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2009 [source] OCCURRENCE OF LISTERIA SPECIES IN THE PROCESSING STAGES OF FROZEN PEPPERJOURNAL OF FOOD SAFETY, Issue 2 2007SOLMAZ LEE ABSTRACT The occurrence of Listeria monocytogenes and other Listeria spp. in a frozen vegetable processing factory was investigated. From May to October 2002, four separate visits were made to the plant and during all of these visits, a total of 216 samples were collected at different stages of the cube and strip pepper processing line. Additionally, 28 swabs were taken from equipment and food-related contact surfaces. The cube and strip pepper processing lines include raw materials, washing, conveyor belt, scalding, cutting, sieving (drying), and the interior sieve of individually quick frozen (IQF), IQF and finished products. Swab samples were taken from the scalding tank, cooling tank, conveyor belt to IQF, interior part of IQF, mixing shovel of IQF, transport saddles and packaging materials. No Listeria spp. were isolated from the strip pepper processing stages, however, 26 out of 108 (24.1%) samples taken from the cube pepper processing stages were found to be contaminated with Listeria spp. Among these isolates, L. monocytogenes was not identified; however, Listeria welshimeri, Listeria innocua and Listeria ivanovii species were identified in 15, 6 and 5 of the tested samples, respectively. L. welshimeri and L. ivanovii were also isolated from three swab samples. These indicate that even though L. monocytogenes was not isolated, the presence of other Listeria species, particularly L. innocua, in the processing line would be an important criterion for eventual L. monocytogenes contaminations. Thus, periodic controls and application of general hygiene and sanitation principles are necessary in the prevention of possible contaminations. [source] SIMULTANEOUS RECOVERY AND DETECTION OF FOUR HEAT-INJURED FOODBORNE PATHOGENS IN GROUND BEEF AND MILK BY A FOUR-COMPARTMENT THIN AGAR LAYER PLATEJOURNAL OF FOOD SAFETY, Issue 2 2006VIVIAN C.H. WU ABSTRACT A four-compartment thin agar layer (4-TAL) system was developed to improve operation efficiency and recover injured foodborne pathogens simultaneously. The system consisted of a layer of nonselective agar overlaid on four different selective agars (xylose lysine desoxycholate [XLD], cefsulodin irgasan novobiocin [CIN], modified Oxford medium [MOX] and MacConkey sorbitol agar [MSA]) housed in a four-compartment petri dish. We applied this system to simultaneously recover heat-injured (55C, 10 min) Escherichia coli O157:H7 (MSA), Listeria monocytogenes (MOX), Salmonella Typhimurium (XLD) and Yersinia enterocolitica (CIN) from ground beef and pasteurized milk. No significant difference (P > 0.05) occurred between the single recovery unit (nonselective agar overlaid on one selective agar in a standard petri dish) and the 4-TAL for detecting four heat-injured pathogens in tested samples. Both TAL methods showed greater recovery of four heat-injured pathogens than the pathogen-specific selective media (P < 0.05). The 4-TAL system appears to be efficient for recovery and detection of injured pathogens in food in terms of operation, material and labor costs, and space of incubation. [source] Trans Fatty Acid Content of Selected Foods in an African-American CommunityJOURNAL OF FOOD SCIENCE, Issue 6 2006Z. Huang ABSTRACT:,Trans fatty acid content was examined in several grocery foods and fast foods in an African-American community. Food samples were selected based on the frequency of use among this population group in the local community. Samples were collected 3 times with an interval of 1 wk. Total fat content was analyzed by Soxhlet method. Fatty acids profile, including trans fatty acids, was analyzed by GC,MS. In grocery foods, no trans fatty acids were detected in fish sticks, salad dressing, mayonnaise, muffin, and potato chips. Margarine contained the highest trans fatty acid at a level of 19.13%. The trans fatty acid level in crackers, cookies, butter, chicken patties, and biscuits mix ranged from 0.51% to 1.77%. In fast foods, no trans fat was detected in dressing. All the fried food and bakery food sampled in this study contain trans fat. The level varied from 2.07% to 10.30%. The principal trans fatty acid was trans 18:1. Other trans fatty acids found were trans 18:2, trans 19:1, and trans 16:1. In a total of 23 food samples, 16 of them were found to contain trans fatty acid. The results demonstrated that trans fat is commonly found in foods of the African-American community. The trans fatty acids content in tested samples varied from 0% to 19.13%. [source] Chemical fingerprint of commercial Radix Echinopsis and quantitative analysis of ,-terthienylJOURNAL OF SEPARATION SCIENCE, JSS, Issue 4-5 2010Xue Qiao Abstract New TLC, HPLC and LC/MS methods were developed for the rapid separation, characterization and quantitation of thiophenes in Radix Echinopsis, a herbal medicine, which has been used in China for long history. Nineteen commercial batches of this herb were analyzed by TLC and HPLC. Only five batches were derived from correct species. The other 14 batches were identified as its adulterating species, Radix Rhapontici, as they did not contain characteristic thiophenes of Echinopsis species. By LC/MS, 16 thiophenes were identified from the methanol extracts of Radix Echinopsis and Radix Rhapontici according to their fragmentation behaviors in MS/MS. In addition, a fully validated HPLC method was established to determine the contents of ,-terthienyl in Radix Echinopsis. The contents ranged from 2.7 to 5.2,mg/g for five tested samples. The methods in this paper were simple and reliable, and could be used for the comprehensive quality control of Radix Echinopsis. [source] Screening food products for the presence of CaMV 35S promoter and NOS 3, terminatorJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 12 2005Hanaa AS Oraby Abstract Biotechnology has enabled the modification of agricultural materials in a very precise way, thereby improving productivity and yields of economically important crops. There are a number of methods available for detecting genetically modified organisms (GMOs). In the present investigation, a qualitative PCR technique has been adopted in order to discriminate between genetically modified and non-modified food products. The qualitative PCR assay employs primers specific for genetic elements that are used to generate genetically engineered agricultural crops. Two of the most common primers used for the detection of GMOs, 35S promoter and NOS 3, terminator, have been tested over a panel of 24 food products purchased from the local market. The results indicated that, out of the 24 food products tested, three products gave positive results with the 35S promoter. The NOS 3, primers gave negative results with all tested samples. Copyright © 2005 Society of Chemical Industry [source] Liquid chromatography/triple quadrupole tandem mass spectrometry with multiple reaction monitoring for optimal selection of transitions to evaluate nutraceuticals from olive-tree materialsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 6 2008Rafael Japón Luján Optimal transitions have been selected for the identification and quantitation of the most interesting hydrophilic biophenols in extracts from olive-tree materials, which are of interest because of their nutraceutical properties. The tested materials were extra virgin olive oil, waste from oil production (known as alperujo), and olive-tree materials such as leaves, small branches and fruit stones. The identification and determination steps of the target biophenols are based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) with a triple quadrupole (QQQ) mass detector. The interface between the chromatograph and the QQQ was an electrospray ionization source operated in the negative ion mode. Highly selective identification of the biophenols was confirmed by multiple reaction monitoring (MRM) using the most representative transitions from the precursor ion to the different product ions. Quantitative MS/MS analysis was carried out by optimization and selection of the most sensitive transition for each analyte, which resulted in estimated detection limits of 5.10 to 11.65,ng/mL for the extracts. The biophenols were extracted from the tested samples by different methods: liquid-liquid extraction for virgin olive oil, microwave-assisted leaching for olive leaves, branches and stones, and pressurized liquid leaching for alperujo. This study provides valuable information about the most suitable source for the isolation of each nutraceutical biophenol and enables us to obtain a complete profile of them in Olea Europaea. Copyright © 2008 John Wiley & Sons, Ltd. [source] Identification of two piwi genes and their expression profile in honeybee, Apis melliferaARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 2 2010Zhen Liao Abstract Piwi genes play an important role in regulating spermatogenesis and oogenesis because they participate in the biogenesis of piRNAs, a new class of noncoding RNAs. However, these genes are not well understood in most insects. To understand the function of piwi genes in honeybee reproduction, we amplified two full-length piwi-like genes, Am-aub and Am-ago3. Both the cloned Am-aub and Am-ago3 genes contained typical PAZ and PIWI domains and active catalytic motifs "Asp-Asp-Asp/His/Glu/Lys," suggesting that the two piwi-like genes possessed slicer activity. We examined the expression levels of Am-aub and Am-ago3 in workers, queens, drones, and female larvae by quantitative PCR. Am-aub was more abundant than Am-ago3 in all the tested samples. Both Am-aub and Am-ago3 were highly expressed in drones but not in workers and queens. The significant finding was that the larval food stream influenced the expression of Piwi genes in adult honeybees. This helps to understand the nutritional control of reproductive status in honeybees at the molecular level. © 2010 Wiley Periodicals, Inc. [source] | |||||||||||||