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pestis fulminans [keywords]
- IpO: Plasmids and methods for simplified, PCR-based DNA transplant in yeast. [JOURNAL ARTICLE]
- Yeast 2014 Mar 6.
Many yeast experiments require strains modified by recombinant DNA methods. Some experiments require precise insertion of a DNA segment into the genome without a selectable marker remaining. For these applications, we developed a new PCR-based method for marker-free DNA transplant. The current PCR-based method requires the labor-intensive construction of a PCR template plasmid with repeats of the DNA segment flanking URA3. The design of a new vector, IpO, reduces the work to cloning a single copy of the DNA segment between overlapping URA3 fragments present in the vector. Two PCRs are performed that capture the DNA segment and one or the other URA3 fragment. When the PCR products are co-transformed into yeast, recombination between the overlapping URA3 fragments restores URA3 and transposes the cloned DNA segment inside out, creating a repeat-URA3-repeat cassette. Sequences designed into the PCR primers target integration of the cassette into the genome. Subsequent selection with 5-fluoro-orotic acid yields strains that have popped out URA3 via recombination between the DNA repeats, with the result being the precise insertion of the DNA segment minus the selectable marker. An additional advantage of the IpO method is that it eliminates PCR artifacts that can plague the current method's repeat-containing templates. This article is protected by copyright. All rights reserved.
- Ancient pathogen DNA in archaeological samples detected with a Microbial Detection Array. [Journal Article]
- Sci Rep 2014.:4245.
Ancient human remains of paleopathological interest typically contain highly degraded DNA in which pathogenic taxa are often minority components, making sequence-based metagenomic characterization costly. Microarrays may hold a potential solution to these challenges, offering a rapid, affordable, and highly informative snapshot of microbial diversity in complex samples without the lengthy analysis and/or high cost associated with high-throughput sequencing. Their versatility is well established for modern clinical specimens, but they have yet to be applied to ancient remains. Here we report bacterial profiles of archaeological and historical human remains using the Lawrence Livermore Microbial Detection Array (LLMDA). The array successfully identified previously-verified bacterial human pathogens, including Vibrio cholerae (cholera) in a 19th century intestinal specimen and Yersinia pestis ("Black Death" plague) in a medieval tooth, which represented only minute fractions (0.03% and 0.08% alignable high-throughput shotgun sequencing reads) of their respective DNA content. This demonstrates that the LLMDA can identify primary and/or co-infecting bacterial pathogens in ancient samples, thereby serving as a rapid and inexpensive paleopathological screening tool to study health across both space and time.
- Modified clay minerals efficiency against chemical and biological warfare agents for civil human protection. [JOURNAL ARTICLE]
- J Hazard Mater 2014 Feb 12.:65-72.
Sorption efficiencies of modified montmorillonite and vermiculite of their mono ionic Na and organic HDTMA and HDP forms were studied against chemical and biological warfare agents such as yperite and selected bacterial strains. Yperite interactions with modified clay minerals were observed through its capture in low-density polyethylene foil-modified clay composites by measuring yperite gas permeation with using chemical indication and gas chromatography methods. The antibacterial activities of synthetized organoclays were tested against selected Gram-positive and Gram-negative bacterial species in minimum inhibitory concentration tests. The obtained results showed a positive influence of modified clay minerals on the significant yperite breakthrough-time increase. The most effective material was the polyethylene-Na form montmorillonite, while the polyethylene-Na form vermiculite showed the lowest efficiency. With increasing organic cations loading in the interlayer space the montmorillonite efficiency decreased, and in the case of vermiculite an opposite effect was observed. Generally the modified montmorillonites were more effective than modified vermiculites. The HDP cations seem to be more effective compare to the HDTMA. The antibacterial activity tests confirmed efficiency of all organically modified clay minerals against Gram-positive bacteria. The confirmation of antibacterial activity against Y. pestis, plague bacteria, is the most interesting result of this part of the study.
- Direct neutralization of type III effector translocation by the variable region of a monoclonal antibody to the LcrV protein of Yersinia pestis. [JOURNAL ARTICLE]
- Clin Vaccine Immunol 2014 Mar 5.
Plague is an acute infection caused by the Gram-negative bacterium Yersinia pestis. Antibodies that are protective against plague target LcrV, an essential virulence protein and component of a type III secretion system of Y. pestis. Secreted LcrV localizes to the tip of type III needles on the bacterial surface and its function is necessary for translocation of Yersinia outer proteins (Yops) into the cytosol of host cells infected by Y. pestis. Translocated Yops counteract macrophage functions, for example, by inhibiting phagocytosis (YopE) or inducing cytotoxicity (YopJ). Although LcrV is the best-characterized protective antigen of Y. pestis, the mechanism of protection by anti-LcrV antibodies is not fully understood. Antibodies bind to LcrV at needle tips, neutralize Yop translocation and promote opsonophagocytosis of Y. pestis by macrophages in vitro. However, it is unclear if LcrV antibodies neutralize Yop translocation directly, or indirectly by promoting opsonophagocytosis. To determine if the protective IgG1 monoclonal antibody (mAb) 7.3 is directly neutralizing, an IgG2a subclass variant, a deglycosylated variant, F(ab')2 and Fab were tested for the ability to inhibit Yop translocation into Y. pestis-infected macrophages in vitro. Macrophage cytotoxicity and cellular fractionation assays show that the Fc of mAb 7.3 is not required to neutralize YopJ or YopE translocation. In addition, use of Fc receptor-deficient macrophages or cytocholasin D to inhibit actin polymerization confirmed that opsonophagocytosis is not required for mAb 7.3 to neutralize translocation. These data indicate that binding of the variable region of mAb 7.3 to LcrV is sufficient to directly neutralize Yop translocation.
- Integrity of scientific content and authorship disputes continue to plague scientific journals. [Journal Article]
- DNA Cell Biol 2014 Mar; 33(3):121.
- A sexually transmitted disease: History of AIDS through philately. [Journal Article]
- J Turk Ger Gynecol Assoc 2011; 12(3):192-6.
AIDS has become the new plague; a disease that is not only physically and psychologically debilitating, but culturally and socially devastating as well. Like the plague, AIDS has caused fear, prejudice and even panic in society. Although there are remarkable improvements in the diagnosis and treatment of the disease, AIDS continues its grim passage around the globe. After a slight downturn in the early 1990's, it then returned with a vengeance. By the end of the 20(th) century, AIDS was reliably estimated to have caused over 20 million deaths throughout the world. At the same time, 40 million people were estimated to be HIV positive. This paper provides an overview of the history of AIDS, including the discovery and its progress in the world through philately. Philately is the study of stamps and postal history and other related items. Philately involves more than just stamp collecting, it contains the study of the design and educational impact of a philatelic material. We have presented AIDS stamps produced world-wide to emphasize the history of AIDS.
- [Suicidal ideation and completed suicide in the decameron]. [English Abstract, Journal Article]
- Turk Psikiyatri Derg 2014; 25(1):38-41.
To extend our understanding of suicidal ideation and completed suicide via examination of a medieval Italian text-in particular, examination of the role of social and environmental triggers in such events. Previous studies have successfully examined the texts of ancient Greco-Roman, Old Norse, and Finnish civilizations to better understand the circumstances associated with suicidal ideation and completed suicide. Those texts included depictions of suicide by people without any apparent mental disorder in response to painful social/environmental circumstances.The Decameron, a collection of 100 short stories written by Giovanni Boccaccio in 1349-51, was examined in detail. The setting of The Decameron is during the Black Death and concerns a group of 10 people that leave plague-ridden Florence for a holiday in the countryside. On each of the 10 days of their trip, each individual tells a story, which in total form the 100 stories of Boccaccio's work. For the present study, all mentions of suicidal ideation and completed suicide were listed and arranged with appropriate headings.In total, 9 accounts of suicidal ideation and 4 accounts of completed suicide were identified, all of which were in response to social stressors, particularly romantic problems.Suicidal ideation and completed suicide in response to social stressors was a feature of The Decameron and it is feasible that this may have been a feature of medieval Italian culture. This would be in agreement with the findings of other studies that suggest that although commonly occurring in response to mental disorder, suicide can occur independently of such disorder as a result of a predicament in which an individual finds him/herself.
- Editorial: Yersinia pestis survives in neutrophils and sends a PS to macrophages: bon appetit! [Journal Article]
- J Leukoc Biol 2014 Mar; 95(3):383-5.
- 'The chearful haunts': John Armstrong (1709-1779), physician, poet, satirist and leveller of medical knowledge. [JOURNAL ARTICLE]
- J Med Biogr 2014 Jan 27.
John Armstrong, the first honours graduate of the University of Edinburgh School of Medicine, was famous in his day for a lengthy didactic poem entitled The Art of Preserving Health (1744). He is now obscure except to scholars specializing in the 18th century and, when discussed at all, often dismissed as a failed physician who wrote mediocre poetry in a quest for money and fame. A new exegesis by Adam Budd exhumes Armstrong as an original voice who offered timely and reassuring advice to Britons as they braced for another epidemic of plague; who depicted illness through the lens of a vulnerable and sympathetic physician, and who was perhaps above all else a leveller of medical knowledge. Elaborating on Budd's thesis, it would seem that Armstrong, a complicated man, has frequently been misread and was in some ways ahead of his time.
- Utilizing DNA analysis to combat the world wide plague of present day slavery - trafficking in persons. [Journal Article]
- Croat Med J 2014 Feb 28; 55(1):3-9.
A study was conducted to determine if modern forensic DNA typing methods can be properly employed throughout the world with a final goal of increasing arrests, prosecutions, and convictions of perpetrators of modern day trafficking in persons while concurrently reducing the burden of victim testimony in legal proceedings. Without interruption of investigations, collection of samples containing DNA was conducted in a variety of settings. Evidentiary samples were analyzed on the ANDE Rapid DNA system. Many of the collected swabs yielded informative short tandem repeat profiles with Rapid DNA technology.