Podcasts about pfge

Apple Pro Display XDRNit’s, also known as Lice - Head Lice - General Information - Frequently Asked QuestionsRapid Response Teams (RRTs) | FDAEcho in the Canyon - WikipediaBarry - Official Website for the HBO SeriesThe Kominsky Method: Season 2 | Official Trailer | Netflix - YouTubeStress Relief (The Office) - WikipediaOcean’s Eleven (2001) - IMDbThe Movies That Made Us (TV Series 2019– ) - IMDbLove Actually - WikipediaFour Weddings and a Funeral - WikipediaTrainspotting (film) - WikipediaFood Safety Talk 7: Dazed and Confused — Food Safety Talk141-year-old fruitcake is a Michigan family’s heirloom - News - Ionia Sentinel - Standard-Ionia, MI - Ionia, MIIs It Safe To Eat Fruitcake While You’re Breastfeeding?Fruitcake – Will it Last Forever? | NC State NewsHoliday foods to avoid during pregnancy | BabyCenterJimmy Buffett - Fruitcakes - YouTubeSquare Dancing (Entry 1211.JB2111) | Omnibus! With Ken Jennings and John Roderick‎You’re Wrong About…: The “Ebonics” Controversy on Apple PodcastsThe Talk Show ✪: Ep. 271, With Special Guest Jason SnellHeavyweight | GimletSafe Plates for Food Managers | NC State ExtensionNotes from the Field: Hospital Water Contamination Associated with a Pseudo-Outbreak of Mycobacterium porcinum — Wisconsin, 2016–2018 | MMWRMultistate Outbreak of Salmonella Infections Linked to Raw Turkey Products — United States, 2017–2019 | MMWRRetrospective whole-genome sequencing analysis distinguished PFGE and drug-resistance-matched retail meat and clinical Salmonella isolates | Microbiology SocietyMultistate Outbreak of Listeriosis Linked to Frozen Vegetables | Listeria | CDCHealth Department Gives Tyson Plant ‘D’ Grade After Discovering Raw Chicken Contaminating Nearly Every SurfaceA Comprehensive Needs Assessment of Food Safety Practices of Farmers’ Market Vendors in Pennsylvania Using Direct Concealed Observations, Self-reported Surveys, and State Sanitarian SurveysPrevalence and Phylogenetic Characterization of Escherichia coli and Hygiene Indicator Bacteria Isolated from Leafy Green Produce, Beef, and Pork Obtained from Farmers’ Markets in Pennsylvania | Journal of Food ProtectionA Comprehensive Food Safety Assessment of Farmers’ Markets in Pennsylvania - DissertationJoshua Scheinberg, Ph.D. | LinkedInSurvey of Food Safety Practices on Small to Medium-Sized Farms and in Farmers Markets | Journal of Food ProtectionAn assessment of food hygiene and safety at farmers’ markets: International Journal of Environmental Health Research: Vol 14, No 2Assessing Food Safety Practices in Farmers’ MarketsFood Safety at Farmers’ Markets: Fact or Fiction? - ScienceDirectMicrobial safety and quality of fresh herbs from Los Angeles, Orange County and Seattle farmers’ markets - Levy - 2015 - Journal of the Science of Food and Agriculture - Wiley Online LibraryIdentification of Risky Food Safety Practices at Southwest Virginia Farmers’ Markets - International Association for Food ProtectionAn assessment of food safety handling practices at farmers’ markets in Rhode Island using a smartphone applicationA Microbiological Survey of Selected Alberta-Grown Fresh Produce from Farmers’ Markets in Alberta, Canada | Journal of Food ProtectionMicrobiological Survey of Locally Grown Lettuce Sold at Farmers’ Markets in Vancouver, British Columbia | Journal of Food ProtectionCross-Sectional Survey of Indicator and Pathogenic Bacteria on Vegetables Sold from Asian Vendors at Farmers’ Markets in Northern California | Journal of Food ProtectionFood Safety? It’s Better Than You ThinkFoodborne Illness Acquired in the United States—Major PathogensNational Outbreak Reporting System (NORS) Dashboard | CDC(5) Larry Stringer (@larry_stringer) / TwitterCommunity engagement and institutional collaboration during outbreaks of Shiga toxin/verocytotoxin- producing Escherichia coli in IrelandCrèche - Wikipedia

The guys started the show following up on the previous episode, the write up on David Gumpert's website and the comments on the Internet. Theresa Lam also reached out wanting to know more about the risks associated with bootleg versus regulated raw milk. Despite raw milk drinker's hatred of epidemiologists, Don confessed that maybe he wants to be an epidemiologist, while Ben noted that he has admired them ever since the Jalapeno Saintpaul outbreak. Don also praised Petran, White and Hedberg, for their efforts to identify what items in a restaurant inspection were predictive of the possibility of an outbreak, and Ben's comments to USA Today on the topic. A quick round of "I think you're thinking of" with Howard Dean, and Roger Dean, not to be confused with Roger Dean followed. The guys then hopped back in time with the whizzinator before moving on to food storage mistakes and tortilla safety as prompted by Listener John Kimble. The guys covered the 1990's in the IAFP history segment, which also featured a discussion of 808, the Beastie Boys and the speed of Joe Walsh's Maserati. Ben identified the 1990's with the adoption of PFGE and rapid methods, while Don though the Mega Regs characterized the time. Ben recalled a recent discussion with Cathy Cutter about meat processing and how HACCP shaped other food safety regulations. The discussion then turned to Norovirus, prompted by a couple of recent noro outbreaks on the "Explorer of the Seas" and the Caribbean Princess, the boat that Chris Gunter boarded. Unfortunately, Chris couldn't find out whether the hand sanitizer on the ship was the one that works, though he was assured that it was "alcohol based". Ben wrapped up the noro discussion with the MoChunk resort outbreak. The guys talked about Netflix in the short after dark.

Due to technical reasons, milk powder and powdered infant formulae (PIF) are not sterile products. In order to achieve the requirements set by the Swiss and European regulations for microbiologic criteria extensive epidemiological studies are needed on each individual plant level. In this way contamination routes can be identified and appropriate measurements taken. A legally reglemented pasteurization process eliminates Enterobacteriaceae. Therefore recontamination must now be focussed on. PIF contaminated with Cronobacter spp. can lead to severe infections in neonates such a sepsis, meningitis or necrotizing enterocolitis. The reported prevalence of commercially available PIF appears to be gradually decreasing from estimates of 14% in 1988 to 2.0-2.5%, where it now seems to have become stabilized. In order to make a reasonable estimate concerning the prevalence of Cronobacter spp. on an individual plant level, 950 samples (raw material, finished products, environmental samples) were analysed. The high prevalence of 16% comes from the intentional sampling of critical raw material and environment samples. The PFGE analysis, however, did not reveal any correlation between raw material and environmental samples which would indicate a possible contamination via finished products. 470 PIF Enterobacteriaceae isolates were identified through biochemical tests as well as by rpoB sequencing. E. cloacae (35%), Pantoea spp. (11%) and K. pneumoniae (8%) were the most prevalent genus and species. In order to reveal possible contamination routes, a subtyping was conducted. The species E. cloacae, which can be found in the same niches as Cronobacter spp., could be used as a significant hygienic indicator organism. To complete the epidemiological picture, 363 milk based samples were analysed (raw milk, milk concentrate, milk powder). Raw milk contains Enterobacteriaceae but no Cronobacter spp. were detected. However, 12/172 samples of milk powder contained Cronobacter spp. due to recontamination (during the packaging process and/or further processing steps). In order to increase the sensitivity and specificity of today’s available analysis for the detection of Cronobacter spp. methodological improvements had to be undertaken. The currently used enrichment media (mLST, EE) contain components of too selective nature which can therefore lead to false negative results. The new “Cronobacter Screening Broth” (CSB) contains sucrose and 5-bromo-4-chloro-3-indolyl-α-D-Glucopyranoside which now leads to a sensitivity of 100% and a negative predicting value of 100% as well. The change in colour of the broth indicates a presumptively positive result whereby only these samples need to be streaked onto chromogenic agar. The visual intermediate result leads to a reduction in costs and working time. In order to increase specificity as well as the commercial pressure of fast product release, a PCR-based system where positive and negative results are clearly available in short time is recommended. Several real-time PCR based systems for detection of Cronobacter spp. have become commercially available. Two systems (one open platform (Biotecon Diagnostics, Potsdam, Germany) and one dedicated system (BioControl, Bellvue, USA)) generated neither false positive nor false negative results. Both systems were able to detect 9 target and 13 non-target strains. The dedicated system has the advantage of shorter hands-on and analysis time. In addition, contaminations due to handling faults are reduced. The existing rpoB based Cronobacter species PCR was upgraded for the recently described species C. condimenti which can now be detected with high reliability. Additional epidemiological data is needed in order to monitor the microbiological situation in industrial plants constantly as well as consequently. Based on information on individual plant level it is possible to implement adequate measurements such as HEPA filters, exact time for adding heat labile ingredients, personal and material flow, air management and cleaning (type, time). Scientific support is needed concerning adequate analytical methods, formation of biofilm, desiccation data, types of enrichment media, sample size as well as additional epidemiological data. Our recent study concerning genetic diversity showed that different Cronobacter isolates from one sample can contain different PFGE fingerprints. This observation suggests that analysis of one isolate per sample may not be sufficient for trace back studies. The analysis of at least five colony forming units per sample is suggested. This example shows that through a close collaboration between industrial companies and scientific institutes, knowledge can be actively turned into practice. – This helps prevent pre-term babies and newborns from falling ill to Cronobacter spp.

Recent work has underlined the importance of animal models in discovery and characterisation of molecular mechanisms determining radiosensitivity and radioresistance. Enhanced sensitivity of LEC rats to ionizing radiation in terms of the acute radiation syndrome was investigated in the present work on the cellular level and compared to that of LE rats. To understand the molecular basis for the increased radiation sensitivity a series of studies were performed, which included the classical clonogenic survival assay, investigation of double strand break repair by means of PFGE and gH2AX evaluation, comet assay for evaluation of repair of single strand breaks and alkaline labile sites, and analysis of cell cycle progression of asynchronous fibroblast population. Survival assay, PFGE, and H2AX analysis were performed in a standardised experimental system - confluent fibroblasts, synchronized in G1 phase of cell cycle, and comet assays were performed in G0 lymphocytes. The data suggests a mild radiosensitization of LEC fibroblasts compared to LE. The results of studies using the selected model did not reflect the degree of animal sensitivity on the molecular level, since values of dose modifying factor (DMF) were much lower in fibroblasts (DMF2 = 1.32) compared to that of animal sensitivity (DMF = 2.36 for bone marrow syndrome (LD50/30) and DMF = 1.95 of intestinal death (LD50/7)). The investigation of DNA repair and cell cycle did not reveal a significant defect in the studied pathways in synchronized fibroblasts and cell cycle progression was not different from wild type cells. The presented data contradict the published LEC cellular phenotype. Of the possible candidate genes, which are located in the radiosensitivity locus, several were further analysed. Among those, Gata-2 appeared to be the most promising of the positional and functional candidates. However, no mutation in the coding sequence could be identified and mRNA expression levels were similar between control and LEC cells. The presented data suggests that radiosensitivity of LEC rats might be attributed to a mechanism specific for certain target tissue, like bone marrow, or enhanced in cell cycle stages other than G0/G1.

Rad5 is a decisive protein in S. cerevisiae due to its role in the Post-replication repair (PRR) pathway, in which Rad5 is necessary for at least one error-free and one error-prone repair subpathway. In addition, Rad5 plays a role in other repair pathways; for instance, Rad5 regulates the balance between the double strand break (DSB) repair pathways, favoring the Rad52-dependent Homologous Recombination (HR) over the yKu70-dependent Non-Homologous-End Joining (NHEJ). Furthermore, since UV-induced damages are substrates for Rad5 but also for Base Excision Repair (BER) proteins, Rad5 is possibly involved directly or indirectly in the BER pathway. To get a deeper insight into the interaction of Rad5 with HR, NHEJ and BER proteins, survival curves, plasmid assays, and mutagenicity experiments were carried out in this work. In addition, a new software tool has been developed for the quantification of DSB. This software, called Geltool, allows the quantification of DSB in haploid cells from PFGE gels, even if the number of DSB is small. This represents a decisive advantage in comparison with previous programs. The sensitivity of Geltool has permitted the quantification of DSB repair during the stationary growth phase in haploid cells, detecting a repair of 46 %- 57 % of the gamma-induced DSB in HR proficient strains against 6 % - 16 % in HR deficient strains. Studies of the functional interactions of Rad5 with HR and NHEJ proteins revealed a synergistic effect between Rad5 and Rad52 proteins for the repair of DSB at chromosomal and plasmidial level. Differences in the repair of plasmids from the rad52 and the rad5 mutants revealed different end joining mechanisms for gap repair. Severe degradations found in plasmids from rad52 and rad52rad5 mutants could indicate an end protection function for Rad52 and also for Rad5, when Rad52 is absent. Moreover, the regulatory role of the Rad5 protein is confirmed, since the additional deletion of YKU70 suppresses the rad5 phenotype and forces the yku70rad5 mutant to repair by HR. The further study of the interplay of Rad5 with BER proteins shows that while BER only plays a minor role for the repair of gamma-induced damage, the rad5 phenotype is suppressed in the BER deficient apn1ntg1ntg2rad5 mutant. The same phenotype of suppression is also found for survival after UV irradiation. An enhanced mutagenicity of the apn1ntg1ntg2rad5 mutant indicates a possible repair through the REV3-dependent Translesion Synthesis Repair (TLS) pathway, suggesting that an error-prone tolerance of UV-induced damage can be very effective for survival.

Current results on the detailed typing of Salmonella Typhimurium from minced meat. The aim of the study was to use PFGE to analyse salmonella isolates from samples of minced meat taken from an EU-approved abattoir and meat processing facility in the period from January 2002 to January 2004. The results were compared with data from previous analyses. Of the total of 350 minced meat samples examined, 36 tested positive for salmonella (10.3 %). The evaluation showed an increase in the number of samples testing positive for salmonella in both summer and winter months. It was remarkable that the samples which most frequently tested positive came from minced meat produced on Tuesdays and Fridays. Detailed typing of the isolates using PFGE was modified and optimised. Only S. Typhimurium and S. Typhimurium var. Copenhagen were examined, as these serovars had been most frequently isolated (51.4 % and 25.7 % respectively), and have a prominent role to play in terms of epidemiology. The macrorestriction enzymes Xba I and Spe I were used. The results following restriction using Xba I were compared with those from previous analyses. Restriction of all isolates using Spe I served to verify the results. Six different restriction patterns were found using Xba I; the majority of the isolates (87.5 %) could be classified in three groups. A comparison with the results from previous analyses showed that two of the groups had been discovered in the minced meat samples of the same facility since 1996. The third group was discovered for the first time in the course of this study. The fact that salmonella isolates with identical macrorestriction patterns have been found in minced meat samples of a production facility over a period of seven years points to a persistent source of contamination which normal hygiene measures have been unable to pick up. Consumer protection would be considerably improved if this source could be detected and eliminated.

Yersinia enterocolitica of bioserovar 4/O:3 is the most common pathogenic type found in pigs and pork including edible offal in Southern Germany. Limited epidemiological data is available of human Y. enterocolitica infections in Bayern. The sources and transmission routes of yersiniosis are not clear. This work was conducted to get more information on the epidemiology in Munich area. In the first part of this work, the human strains were isolated from faecal samples of patients with diarrhoea during the period from November 2001 to December 2002. Yersinia was isolated from the faeces using direct culturing on selective CIN agar plates. One typical colony per sample was identified using EnterotubeTM. All Yersinia strains were bio- and serotyped. In all, 61 Yersinia strains were isolated from 61 patients. Fifty-eight (95%) strains were identified as Y. enterocolitica of which 54 (93%) were belonging to pathogenic bieserovar 4/O:3, two (3%) strains to pathogenic bioserovar 2/O:9 and two to non-pathogenic biovar 1A. In 2002, Yersinia was found from 50 (0,21%) patients when 22.835 faeces samples were studied. Y. enterocolitica was identified in 48 (96%) cases. The bioserovar 4/O:3 was found in 44 (92%) patients and bioserovar 2/O:9 in two (4%) patient. Y. enterocolitica was found in all age groups and 60% of the patients were females. No seasonal variation was seen. Yersinia was the third most common bacterial pathogen after Campylobacter found in 805 (3,52%) faecal samples and Salmonella found in 681 (2,98%) samples. In the second part of this work, 36 human and 69 non-human Yersinia strains were characterised with PFGE. The human strains were isolated during the period from November 2001 to June 2002. The non-human strains were isolated between 1999 and 2002. Most (62/69) of these strains were from porcine origin. In total, 41 genotypes were obtained when the 105 strains were characterised with PFGE using NotI, ApaI and XhoI enzymes. The Yersinia spp. could easily be distinguished from each other with NotI enzyme. Twenty-four genotypes were obtained when the 84 strains belonging to bioserovar 4/O:3 was characterised with all three enzymes. Most (90%) of the human strains were indistinguishable from the non-human strains. These 26 indistinguishable human strains were belonging to 8 genotypes. The non-human strains were from porcine sources and isolated from carcasses, offal and pork. These results demonstrate that Y. enterocolitica is frequently isolated from patients with diarrhoea in Munich area. The most common type in humans is the bioserovar 4/O:3, which is so far the only pathogenic type found in pigs in Southern Germany. PFGE with NotI enzyme was useful tool to identify Yersinia spp. Using NotI, ApaI and XhoI enzymes, the strains of bioserovar 4/O:3 could efficiently be subtyped. A major infection source of human yersiniosis was revealed to be pork and edible offal.

Plastid chromosomes from the variety of plant species contain several conserved open reading frames of unknown function, which most probably represent functional genes. The primary aim of this thesis was the analysis of the role of two such ORFs, designated ycfs or hypothetical chloroplast reading frames, namely ycf9 (ORF62) and ycf10 (ORF229, cemA). Both were analyzed in Nicotiana tabacum (tobacco) via their inactivation using biolistic plastid transformation. A new experimental protocol, based on pulsed-field gel electrophoresis (PFGE), was established to reliably assess the homoplastomic state of transformed plants. 1. Functional analysis of the ycf9 gene product: The inactivation of ycf9 in N. tabacum as well as in Chlamydomonas reinhardtii yielded a homoplastomic mutant phenotype after several rounds of regeneration under selective pressure. The mutant plants grew photoautotrophically, but displayed two clear phenotypes, a light-sensitive one, increasing with the light intensity, and a dwarf phenotype under low-light combined with temperatures below 20°C. The ycf9 gene product was exclusively located in PSII core complexes. This localization was based on the isolation of protein complexes released from thylakoids by controlled, partial lysis, followed by sucrose density gradient centrifugation or 2D gel electrophoresis. This finding revised data of the literature. Biochemical analysis indicated an involvement of the protein in the interaction of the light harvesting antenna II complex (LHCII) with PSII cores. In particular, PSII-LHCII supercomplexes could no longer be isolated from transplastomic tobacco plants. Furthermore, the minor chlorophyll a/b-binding proteins CP26, and to a lesser extent CP29, were substantially reduced under most growth conditions analyzed, in both, tobacco and photoautotrophically grown Chlamydomonas mutants (Swiatek et al. 2001). The gene was therefore renamed psbZ. The ∆psbZ-related alterations in the supramolecular organization of PSII complexes were accompanied by considerable modification in (i) the phosphorylation pattern of PSII subunits, (ii) the rate of deepoxydation of xanthophylls, and (iii) the kinetics and amplitude of non-photochemical quenching. The proposed position of PsbZ in close proximity to CP43 enables the protein to interact with PSII cores to elicit an adaptation process in response to excess light excitation. The molecular mechanism underlying this energy dissipation process remains to be investigated. 2. Functional analysis of the ycf10 gene product: Biolistic plastid transformation was also used to inactivate the ycf10 reading frame in tobacco. After several rounds of regeneration under selective pressure, homoplastomic plants were obtained. Northern analysis uncovered co-transcription of ycf10 within the psaI-ycf4-ycf10-petA gene cluster, with at least two promotor regions upstream of the psaI gene. The mutant plants grew photoautotrophically and developed dark green leaves with numerous pale green to white regions, the latter devoid of photosynthetic activity. The loss of ycf10 did not affect photosynthetic activity, as indicated by unaltered chlorophyll fluorescence. The tobacco ycf10 gene product was localized in the chloroplast inner envelope membrane. Neither protein composition of stroma or thylakoid fractions, nor the stability of the photosynthetic protein complexes were affected in the mutant plants. In contrast, CO2- dependent oxygen evolution was strongly reduced, with a maximum rate of Ci-dependent photosynthesis being approximately 50% lower than in wild-type plants. Two explanations can account for the observed phenomenon: (i) de-regulation of carbonconcentrating mechanisms in transformed cells, or (ii) an indirect effect on CO2-uptake in ∆ycf10 plants. 3. Pulsed-field gel electrophoresis is an ideal tool to verify the homoplastomic state of transformed plants: To enhance the sensitivity of detection of heteroplastomic states, and to distinguish between plastome-located wild-type segments in transplastomic material and promiscuous DNA, a new approach was developed. Customary Southern and PCR techniques are not sensitive enough or not discriminating the latter alternatives, respectively. Pulsed-field gel electrophoresis allows to isolate virtually contamination-free plastid DNA. Plastid DNA isolated this way lacked traces of nuclear and mitochondrial DNA at a detection level of 50 DNA molecules. This excludes that gene-specific PCR amplification products originate from promiscuous nuclear or mitochondrial gene copies. Therefore, PFGE appears to be an ideal tool to investigate the homoplastomic state of transformed plants, especially when combined with radiolabeled probes and Southern techniques.

A procedure for the quantification of double-strand breaks in yeast is presented that utilizes pulsed field gel electrophoresis (PFGE) and a comparison of the observed DNA mass distribution in the gel lanes with calculated distributions. Calculation of profiles is performed as follows. If double-strand breaks are produced by sparsely ionizing radiation, one can assume that they are distributed randomly in the genome, and the resulting DNA mass distribution in molecular length can be predicted by means of a random breakage model. The input data for the computation of molecular length profiles are the breakage frequency per unit length, , as adjustable parameter, and the molecular lengths of the intact chromosomes. The obtained DNA mass distributions in molecular length must then be transformed into distributions of DNA mass in migration distance. This requires a calibration of molecular length vs. migration distance that is specific for the gel lane in question. The computed profiles are then folded with a Lorentz distribution with adjusted spread parameter to account for and broadening. The DNA profiles are calculated for different breakage frequencies and for different values of , and the parameters resulting in the best fit of the calculated to the observed profile are determined.