84 research outputs found
Defining Mob Grazing in the Upper Midwestern United States
Mob grazing has emerged as an increasingly used management strategy on pasture-based farms throughout the country; however, the practice lacks clear definition among practitioners. We conducted a survey of livestock and dairy producers using some form of rotational grazing in the upper midwestern United States (N = 155) to gather producer-generated definitions, perceptions of benefits and disadvantages, and implementation strategies for mob grazing. The results describe a practice defined by variability and associated with compelling impacts. Implementation of mob grazing differed among producers, although most used it as a strategic tool and not a rigid management strategy
Surface-Exposed Adeno-Associated Virus Vp1-NLS Capsid Fusion Protein Rescues Infectivity of Noninfectious Wild-Type Vp2/Vp3 and Vp3-Only Capsids but Not That of Fivefold Pore Mutant Virions
Over the past 2 decades, significant effort has been dedicated to the development of adeno-associated virus (AAV) as a vector for human gene therapy. However, understanding of the virus with respect to the functional domains of the capsid remains incomplete. In this study, the goal was to further examine the role of the unique Vp1 N terminus, the N terminus plus the recently identified nuclear localization signal (NLS) (J. C. Grieger, S. Snowdy, and R. J. Samulski, J. Virol 80:5199-5210, 2006), and the virion pore at the fivefold axis in infection. We generated two Vp1 fusion proteins (Vp1 and Vp1NLS) linked to the 8-kDa chemokine domain of rat fractalkine (FKN) for the purpose of surface exposure upon assembly of the virion, as previously described (K. H. Warrington, Jr., O. S. Gorbatyuk, J. K. Harrison, S. R. Opie, S. Zolotukhin, and N. Muzyczka, J. Virol 78:6595-6609, 2004). The unique Vp1 N termini were found to be exposed on the surfaces of these capsids and maintained their phospholipase A2 (PLA2) activity, as determined by native dot blot Western and PLA2 assays, respectively. Incorporation of the fusions into AAV type 2 capsids lacking a wild-type Vp1, i.e., Vp2/Vp3 and Vp3 capsid only, increased infectivity by 3- to 5-fold (Vp1FKN) and 10- to 100-fold (Vp1NLSFKN), respectively. However, the surface-exposed fusions did not restore infectivity to AAV virions containing mutations at a conserved leucine (Leu336Ala, Leu336Cys, or Leu336Trp) located at the base of the fivefold pore. EM analyses suggest that Leu336 may play a role in global structural changes to the virion directly impacting downstream conformational changes essential for infectivity and not only have local effects within the pore, as previously suggested
The effects of preoperative chemotherapy on isolated tumour cells in the blood and bone marrow of gastric cancer patients
Recent studies in breast cancer suggest that monitoring the isolated tumour cells (ITC) may be used as a surrogate marker to evaluate the efficacy of systemic chemotherapy. In the present study, we have investigated the effects of preoperative chemotherapy on ITC in the blood and bone marrow of patients with potentially resectable gastric cancer. After sorting out the CD45-positive cells, the presence of ITC defined as cytokeratin-positive cells was examined before and after preoperative chemotherapy. The patients received two courses of preoperative chemotherapy with cisplatin (100 mg m−2, day 1) and 5-fluorouracil (1000 mg m−2, days 1–5), administered every 28 days. Fourteen of 32 (44%) patients initially diagnosed with ITC in blood and/or bone marrow were found to be negative (responders) after preoperative chemotherapy (P<0.01). The incidence of ITC in bone marrow was also significantly (P<0.01) reduced from 97 (31 of 32) to 53% (17 of 32). The difference between patients positive for ITC in the blood before (n=7, 22%) and after (n=5, 16%) chemotherapy was statistically insignificant. The overall 3-year survival rates were 32 and 49% in the responders and non-responders, respectively (P=0.683). These data indicate that preoperative chemotherapy can reduce the incidence of ITC in patients with gastric cancer
Procalcitonin for diagnosis of infection and guide to antibiotic decisions: past, present and future
There are a number of limitations to using conventional diagnostic markers for patients with clinical suspicion of infection. As a consequence, unnecessary and prolonged exposure to antimicrobial agents adversely affect patient outcomes, while inappropriate antibiotic therapy increases antibiotic resistance. A growing body of evidence supports the use of procalcitonin (PCT) to improve diagnosis of bacterial infections and to guide antibiotic therapy. For patients with upper and lower respiratory tract infection, post-operative infections and for severe sepsis patients in the intensive care unit, randomized-controlled trials have shown a benefit of using PCT algorithms to guide decisions about initiation and/or discontinuation of antibiotic therapy. For some other types of infections, observational studies have shown promising first results, but further intervention studies are needed before use of PCT in clinical routine can be recommended. The aim of this review is to summarize the current evidence for PCT in different infections and clinical settings, and discuss the reliability of this marker when used with validated diagnostic algorithms
Capsid Antibodies to Different Adeno-Associated Virus Serotypes Bind Common Regions
Interactions between viruses and the host antibody immune response are critical in the development and control of disease, and antibodies are also known to interfere with the efficacy of viral vector-based gene delivery. The adeno-associated viruses (AAVs) being developed as vectors for corrective human gene delivery have shown promise in clinical trials, but preexisting antibodies are detrimental to successful outcomes. However, the antigenic epitopes on AAV capsids remain poorly characterized. Cryo-electron microscopy and three-dimensional image reconstruction were used to define the locations of epitopes to which monoclonal fragment antibodies (Fabs) against AAV1, AAV2, AAV5, and AAV6 bind. Pseudoatomic modeling showed that, in each serotype, Fabs bound to a limited number of sites near the protrusions surrounding the 3-fold axes of the T=1 icosahedral capsids. For the closely related AAV1 and AAV6, a common Fab exhibited substoichiometric binding, with one Fab bound, on average, between two of the three protrusions as a consequence of steric crowding. The other AAV Fabs saturated the capsid and bound to the walls of all 60 protrusions, with the footprint for the AAV5 antibody extending toward the 5-fold axis. The angle of incidence for each bound Fab on the AAVs varied and resulted in significant differences in how much of each viral capsid surface was occluded beyond the Fab footprints. The AAV-antibody interactions showed a common set of footprints that overlapped some known receptor-binding sites and transduction determinants, thus suggesting potential mechanisms for virus neutralization by the antibodies
AAV ancestral reconstruction library enables selection of broadly infectious viral variants
Adeno-associated virus (AAV) vectors have achieved clinical efficacy in treating several diseases. Enhanced vectors are required to extend these landmark successes to other indications, however, and protein engineering approaches may provide the necessary vector improvements to address such unmet medical needs. To generate new capsid variants with potentially enhanced infectious properties, and to gain insights into AAV’s evolutionary history, we computationally designed and experimentally constructed a putative ancestral AAV library. Combinatorial variations at 32 amino acid sites were introduced to account for uncertainty in their identities. We then analyzed the evolutionary flexibility of these residues, the majority of which have not been previously studied, by subjecting the library to iterative selection on a representative cell line panel. The resulting variants exhibited transduction efficiencies comparable to the most efficient extant serotypes, and in general ancestral libraries were broadly infectious across the cell line panel, indicating that they favored promiscuity over specificity. Interestingly, putative ancestral AAVs were more thermostable than modern serotypes and did not utilize sialic acids, galactose, or heparan sulfate proteoglycans for cellular entry. Finally, variants mediated 19–31 fold higher gene expression in muscle compared to AAV1, a clinically utilized serotype for muscle delivery, highlighting their promise for gene therapy
System integracji semantycznej nieustrukturyzowanych dokumentów w języku naturalnym z zakresu metalurgii
This paper presents assumptions for a system of automatic cataloging and semantic text documents searching. As an example, a document repository for metals processing technology was used. The system by using ontological model provides the user with a new approach to the exploration of database resources - easier and more intuitive information search. In the current document storage systems, searching is often based only on keywords and descriptions created manually by the system administrator. The use of text mining methods, especially latent semantic indexing, allows automatic clustering of documents with respect to their content. The result of this clustering is integrated with the ontological model, making navigation through documents resources intuitive and does not require the manual creation of directories. Such an approach seems to be particularly useful in a situation where we are dealing with large repositories of unstructured documents from such sources as the Internet. This situation is very typical for cases of searching information and knowledge in the area of metallurgy, for example with regard to innovation and non-traditional suppliers of materials and equipment.Artykuł prezentuje założenia systemu umożliwiającego automatyczne katalogowanie i przeszukiwanie merytoryczne dokumentów tekstowych na przykładzie repozytorium dokumentów dotyczących technologii przetwórstwa metali. System dzięki zastosowaniu modelu ontologicznego ma umożliwić użytkownikowi nowe podejście do eksploracji zasobów bazodanowych - prostsze i bardziej intuicyjne wyszukiwanie informacji. W obecnych systemach przechowywania dokumentów często jedyna forma wyszukiwania jest wyszukiwanie na podstawie katalogu słów kluczowych i deskrypcji tworzonych ręcznie przez administratora systemu. Zastosowanie metod eksploracji tekstu, w szczególności ukrytego indeksowania semantycznego umożliwia automatyczne grupowanie dokumentów pod względem ich zawartości. Wynik takiego grupowania zostaje zintegrowany z modelem ontologicznym, przez co nawigacja poprzez zasoby dokumentów staje się intuicyjna i nie wymaga tworzenia ręcznie katalogów. Takie podejście wydaje się szczególnie przydatne w sytuacji, gdy mamy do czynienia z dużymi repozytoriami nieuporzadkowanych dokumentów pochodzących m.in. z sieci Internet
Total alcaline phosphatase and its placental isoenzyme activity during pregnancy, delivery and confinement
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