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Sample records for rapidly growing human

  1. Mycobacterium grossiae sp. nov., a rapidly growing, scotochromogenic species isolated from human clinical respiratory and blood culture specimens.

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    Paniz-Mondolfi, Alberto Enrique; Greninger, Alexander L; Ladutko, Lynn; Brown-Elliott, Barbara A; Vasireddy, Ravikiran; Jakubiec, Wesley; Vasireddy, Sruthi; Wallace, Richard J; Simmon, Keith E; Dunn, Bruce E; Jackoway, Gary; Vora, Surabhi B; Quinn, Kevin K; Qin, Xuan; Campbell, Sheldon

    2017-11-01

    A previously undescribed, rapidly growing, scotochromogenic species of the genus Mycobacterium (represented by strains PB739 T and GK) was isolated from two clinical sources - the sputum of a 76-year-old patient with severe chronic obstructive pulmonary disease, history of tuberculosis exposure and Mycobacterium avium complex isolated years prior; and the blood of a 15-year-old male with B-cell acute lymphoblastic leukaemia status post bone marrow transplant. The isolates grew as dark orange colonies at 25-37 °C after 5 days, sharing features in common with other closely related species. Analysis of the complete 16S rRNA gene sequence (1492 bp) of strain PB739 T demonstrated that the isolate shared 98.8 % relatedness with Mycobacterium wolinskyi. Partial 429 bp hsp65 and 744 bp rpoB region V sequence analyses revealed that the sequences of the novel isolate shared 94.8 and 92.1 % similarity with those of Mycobacterium neoaurum and Mycobacterium aurum, respectively. Biochemical profiling, antimicrobial susceptibility testing, HPLC/gas-liquid chromatography analyses and multilocus sequence typing support the taxonomic status of these isolates (PB739 T and GK) as representatives of a novel species. Both isolates were susceptible to the Clinical and Laboratory Standards Institute recommended antimicrobials for susceptibility testing of rapidly growing mycobacteria including amikacin, ciprofloxacin, moxifloxacin, doxycycline/minocycline, imipenem, linezolid, clarithromycin and trimethropin/sulfamethoxazole. Both isolates PB739 T and GK showed intermediate susceptibility to cefoxitin. We propose the name Mycobacterium grossiae sp. nov. for this novel species and have deposited the type strain in the DSMZ and CIP culture collections. The type strain is PB739 T (=DSM 104744 T =CIP 111318 T ).

  2. Mycobacterium oryzae sp. nov., a scotochromogenic, rapidly growing species is able to infect human macrophage cell line.

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    Ramaprasad, E V V; Rizvi, A; Banerjee, S; Sasikala, Ch; Ramana, Ch V

    2016-11-01

    Gram-stain-positive, acid-fast-positive, rapidly growing, rod-shaped bacteria (designated as strains JC290T, JC430 and JC431) were isolated from paddy cultivated soils on the Western Ghats of India. Phylogenetic analysis placed the three strains among the rapidly growing mycobacteria, being most closely related to Mycobacterium tokaiense 47503T (98.8 % 16S rRNA gene sequence similarity), Mycobacterium murale MA112/96T (98.8 %) and a few other Mycobacterium species. The level of DNA-DNA reassociation of the three strains with M. tokaiense DSM 44635T was 23.4±4 % (26.1±3 %, reciprocal analysis) and 21.4±2 % (22.1±4 %, reciprocal analysis). The three novel strains shared >99.9 % 16S rRNA gene sequence similarity and DNA-DNA reassociation values >85 %. Furthermore, phylogenetic analysis based on concatenated sequences (3071 bp) of four housekeeping genes (16S rRNA, hsp65, rpoB and sodA) revealed that strain JC290T is clearly distinct from all other Mycobacteriumspecies. The three strains had diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositolmannosides, unidentified phospholipids, unidentified glycolipids and an unidentified lipid as polar lipids. The predominant isoprenoid quinone for all three strains was MK-9(H2). Fatty acids were C17 : 1ω7c, C16 : 0, C18 : 1ω9c, C16 : 1ω7c/C16 : 1ω6c and C19 : 1ω7c/C19 : 1ω6c for all the three strains. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it was concluded that strains JC290T, JC430 and JC431 are members of a novel species within the genus Mycobacterium and for which the name Mycobacterium oryzae sp. nov. is proposed. The type strain is JC290T (=KCTC 39560T=LMG 28809T).

  3. A Multi-Level Approach to Modeling Rapidly Growing Mega-Regions as a Coupled Human-Natural System

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    Koch, J. A.; Tang, W.; Meentemeyer, R. K.

    2013-12-01

    The FUTure Urban-Regional Environment Simulation (FUTURES) integrates information on nonstationary drivers of land change (per capita land area demand, site suitability, and spatial structure of conversion events) into spatial-temporal projections of changes in landscape patterns (Meentemeyer et al., 2013). One striking feature of FUTURES is its patch-growth algorithm that includes feedback effects of former development events across several temporal and spatial scales: cell-level transition events are aggregated into patches of land change and their further growth is based on empirically derived parameters controlling its size, shape, and dispersion. Here, we augment the FUTURES modeling framework by expanding its multilevel structure and its representation of human decision making. The new modeling framework is hierarchically organized as nested subsystems including the latest theory on telecouplings in coupled human-natural systems (Liu et al., 2013). Each subsystem represents a specific level of spatial scale and embraces agents that have decision making authority at a particular level. The subsystems are characterized with regard to their spatial representation and are connected via flows of information (e.g. regulations and policies) or material (e.g. population migration). To provide a modeling framework that is applicable to a wide range of settings and geographical regions and to keep it computationally manageable, we implement a 'zooming factor' that allows to enable or disable subsystems (and hence the represented processes), based on the extent of the study region. The implementation of the FUTURES modeling framework for a specific case study follows the observational modeling approach described in Grimm et al. (2005), starting from the analysis of empirical data in order to capture the processes relevant for specific scales and to allow a rigorous calibration and validation of the model application. In this paper, we give an introduction to the basic

  4. Management of Infections with Rapidly Growing

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    Jong Hwan Kim

    2012-01-01

    Full Text Available Background Infection caused by rapidly growing mycobacteria (RGM is not uncommon, andthe prevalence of RGM infection has been increasing. Clinical diagnosis is difficult becausethere are no characteristic clinical features. There is also no standard antibiotic regimenfor treating RGM infection. A small series of patients with RGM infections was studied toexamine their treatments and outcomes.Methods A total of 5 patients who had developed postoperative infections from January2009 to December 2010 were retrospectively reviewed. Patients were initially screened using amycobacteria rapid screening test (polymerase chain reaction [PCR]-reverse blot hybridizationassay. To confirm mycobacterial infection, specimens were cultured for nontuberculousmycobacteria and analyzed by 16 S ribosomal RNA and rpoB gene PCR.Results The patients were treated with intravenous antibiotics during hospitalization,and oral antibiotics were administered after discharge. The mean duration of follow-upwas 9 months, and all patients were completely cured of infection with a regimen of acombination of antibiotics plus surgical treatment. Although none of the patients developedrecurrence, there were complications at the site of infection, including hypertrophic scarring,pigmentation, and disfigurement.Conclusions Combination antibiotic therapy plus drainage of surgical abscesses appeared tobe effective for the RGM infections seen in our patients. Although neither the exact dosagenor a standardized regimen has been firmly established, we propose that our treatment canprovide an option for the management of rapidly growing mycobacterial infection.

  5. Neonatal airway obstruction caused by rapidly growing nasopharyngeal teratoma.

    NARCIS (Netherlands)

    Maartens, I.A.; Wassenberg, T.; Halbertsma, F.J.; Marres, H.A.M.; Andriessen, P.

    2009-01-01

    A case report is presented of a rapidly growing congenital nasopharyngeal teratoma (epignathus) in a preterm infant, leading to severe upper airway obstruction. Prenatal diagnosis by ultrasonography did not reveal the condition because the tumour masses were initially small and there was no

  6. Environmental and Social Programmes and Rapidly Growing Retailers

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    Peter JONES

    2014-06-01

    Full Text Available This paper looks to provide an exploratory review of the extent to which the world’s fastest growing retailers are publicly reporting on their environmental and social commitments and programmes. The paper begins with an outline discussion of corporate environmental and social programmes and on public reporting processes. The paper draws its empirical material from the most recent information on environmental and social commitments and programmes posted on the world’s top twenty fastest growing retailers’ corporate web sites. While the majority of the world’s top twenty fastest growing retailers provide some public information on their commitment to environmental and social programmes there is marked variation in the extent, the nature and the detail of that information. The findings suggest that the integration of environmental and social programmes is not one of the hallmarks of rapidly growing retailers and in part this reflects the fact that many of the selected retailers are trading within emergent markets where price and availability are the principal factors driving consumer buying behaviour.

  7. Biomass accumulation in rapidly growing loblolly pine and sweetgum

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Thomas M.; Gresham, Charles A. [Baruch Institute of Coastal Ecology and Forest Science, Clemson University, P.O. Box 596, Georgetown, SC 29442 (United States)

    2006-04-15

    Loblolly pine (Pinus taeda) and sweetgum (Liquidambar styraciflua) trees, growing in International Paper Company's study of intensive management on marginal agricultural land near Bainbridge GA, were destructively sampled at the end of the sixth growing season. All trees were single family blocks of genetically superior trees planted 2.5m apart on sub-soiled rows 3.6m apart and grown with complete competition control. Management treatments were: control, irrigation, irrigation plus fertilization, and irrigation plus fertilization plus pest control. Tree measures were basal diameter, DBH, height of live crown, diameter at base of live crown, and total height. Twenty trees of each species were destructively sampled. Stems were sectioned at 1m intervals, stem diameter determined at each end and sections were weighed green. Branches were removed and height, basal diameter, and length were measured on each branch. Branches were separated into foliated and unfoliated segments and weighed green. A stem disk and branch from each meter were returned to the lab to determine dry weight: green weight ratio. Foliated limb: foliage ratios were also determined from sub-sampled branches. Intensive culture resulted in larger growth differences for sweetgum (most intensive treatment 9.5m tall, 13.1cm DBH; control trees 5.0m tall, 6.3cm DBH) than in pine (most intensive treatment 10.3m tall, 17.7cm DBH; control, 7.6m tall, 13.4cm DBH). The pipe model of tree development explained dimensions of the upper 5m of crown with leaf biomass highly correlated to branch basal area (r{sup 2} from 0.697 to 0.947). There was a constant ratio of leaf biomass to branch basal area (50gm/cm{sup 2} for pine, 30gm/cm{sup 2} for sweetgum). We also found a constant ratio of bole basal area to cumulative branch basal area throughout the crowns. Rapidly growing pines produced about 49Mgha{sup -1} of stem biomass, 11Mgha{sup -1} of dead branch biomass, and 17Mgha{sup -1} of unfoliated branch biomass at

  8. Rapidly Growing Esophageal Carcinosarcoma Reduced by Neoadjuvant Radiotherapy Alone

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    Naotaka Ogasawara

    2014-06-01

    Full Text Available Esophageal carcinosarcoma is a rare malignant neoplasm consisting of both carcinomatous and sarcomatous components. It is generally treated by surgery, radiotherapy and chemotherapy according to the protocols used for other esophageal cancers. However, the treatment of esophageal carcinosarcoma by radiotherapy alone before surgery has not been previously described. We report a patient with a rapidly growing esophageal carcinosarcoma that was efficiently reduced by neoadjuvant radiotherapy alone. A previously healthy 69-year-old man was admitted with dysphagia. Initial esophagogastroduodenoscopy (EGD revealed a small nodular polypoid lesion of about 10 mm in the middle esophagus. A second EGD 1 month later showed that the tumor had expanded into a huge mass. A biopsy specimen revealed that the tumor comprised squamous cell carcinoma with spindle cell components, and the tumor was diagnosed as carcinosarcoma which was diagnosed as stage I (T1bN0M0. Due to renal dysfunction, the patient was treated with neoadjuvant radiotherapy (40 Gy without chemotherapy. A third EGD 1 month later revealed remarkable tumor reduction. He then underwent total esophagectomy with regional lymph node dissection (pStage 0, pT1aN0M0. After surgical operation, the patient was followed up without adjuvant therapy. Whole body computed tomography revealed lung metastasis 14 months after surgery, and the patient died 2 months later. The neoadjuvant radiotherapy for esophageal carcinosarcoma was considered to have contributed to the subsequent surgery and his prolonged survival time. Thus, radiotherapy alone might be a suitable neoadjuvant therapy for esophageal carcinosarcomas.

  9. Intraoral tumor with rapid growing. Report of a case.

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    González-Martín-Moro, Javier; Cebrián-Carretero, Jose Luis; Gómez-García, Elena; del Castillo-Pardo de Vera, Jose Luis; del Val, Daniel

    2005-01-01

    The appearance of an intraoral mass is common in our specialty. Most are benign lesions, but some are primary malignancies. Metastases account for less than 1% of all oral malignancies. An 86 year old woman was referred to our department with a large, asymptomatic, intraoral, fast-growing mass. She had no previous cancer history or other relevant physical findings. The radiology studies showed underlying bone erosion. The histological study showed a metastatic adenocarcinoma with a suspected origin in the abdomen. We were unable to identify it by non invasive diagnostic procedures. Given the patient's general status and despite the ominous prognosis of such lesions, we decided not to perform any aggressive therapy beyond removing the oral mass, in order to maintain her quality of life. There have been no local recurrences until this time.

  10. Neuropsychology of music – a rapidly growing branch of psychology

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    Katarina Habe

    2010-05-01

    Full Text Available Relationship between brain and music is of interest to musicians, psychologists and neuroscientists. In recent years no other area of psychology of music has seen as much advancement as neuropsychology of music. The aim of the article is to present some main issues in the neuropsychology of music abroad and in Slovenia, to classify research studies into larger categories and to predict the future development of this field.There are different levels of inquiry into the neuropsychology of music: (1 the analysis of normal and abnormal psychological and physiological functions to determine the principles and modes by which the human brain processes, codifies, stores, and produces music, and (2 a description of the clinical deficits in music perception or performance resulting from localized or diffuse damage to the nervous system. Main topics that occupy neuropsychology of music are neuropsychological models of musical processing, functional imaging of musical perception and cognition, and the use of music as a therapeutic and clinical tool. Although some important studies have already been conducted since the year 2003, in Slovenia we faced a "formal" turning point in acknowledging the importance of the connection between music, mind and brain with the Sinapsa's Week of the brain 2009 under the title Brain and music.

  11. Frequency of rapid growing mycobacteria among tuberculosis suspected patients in Basra-Iraq

    National Research Council Canada - National Science Library

    Sulami, Amin A. Al; Taee, Asaad Al; Hasan, Zainab A

    2016-01-01

    Objective: The purpose of this study was to estimate the frequency of rapid growing mycobacteria among tuberculosis suspected patients in Basra governorate and study their resistance to drugs. Methods...

  12. Aquaculture: a rapidly growing and significant source of sustainable food? Status, transitions and potential.

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    Little, D C; Newton, R W; Beveridge, M C M

    2016-08-01

    The status and potential of aquaculture is considered as part of a broader food landscape of wild aquatic and terrestrial food sources. The rationale and resource base required for the development of aquaculture are considered in the context of broader societal development, cultural preferences and human needs. Attention is drawn to the uneven development and current importance of aquaculture globally as well as its considerable heterogeneity of form and function compared with established terrestrial livestock production. The recent drivers of growth in demand and production are examined and the persistent linkages between exploitation of wild stocks, full life cycle culture and the various intermediate forms explored. An emergent trend for sourcing aquaculture feeds from alternatives to marine ingredients is described and the implications for the sector with rapidly growing feed needs discussed. The rise of non-conventional and innovative feed ingredients, often shared with terrestrial livestock, are considered, including aquaculture itself becoming a major source of marine ingredients. The implications for the continued expected growth of aquaculture are set in the context of sustainable intensification, with the challenges that conventional intensification and emergent integration within, and between, value chains explored. The review concludes with a consideration of the implications for dependent livelihoods and projections for various futures based on limited resources but growing demand.

  13. ISOLATION AND ANTIBIOTIC SUSCEPTIBILITY TESTING OF RAPIDLY-GROWING MYCOBACTERIA FROM GRASSLAND SOILS

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    Martina Kyselková

    2013-08-01

    Full Text Available Rapidly growing mycobacteria (RGM are common soil saprophytes, but certain strains cause infections in human and animals. The infections due to RGM have been increasing in past decades and are often difficult to treat. The susceptibility to antibiotics is regularly evaluated in clinical isolates of RGM, but the data on soil RGM are missing. The objectives of this study was to isolate RGM from four grassland soils with different impact of manuring, and assess their resistance to antibiotics and the ability to grow at 37°C and 42°C. Since isolation of RGM from soil is a challenge, a conventional decontamination method (NaOH/malachite green/cycloheximide and a recent method based on olive oil/SDS demulsification were compared. The olive oil/SDS method was less efficient, mainly because of the emulsion instability and plate overgrowing with other bacteria. Altogether, 44 isolates were obtained and 23 representatives of different RGM genotypes were screened. The number of isolates per soil decreased with increasing soil pH, consistently with previous findings that mycobacteria were more abundant in low pH soils. Most of the isolates belonged to the Mycobacterium fortuitum group. The majority of isolates was resistant to 2-4 antibiotics. Multiresistant strains occurred also in a control soil that has a long history without the exposure to antibiotic-containing manure. Seven isolates grew at 37°C, including the species M. septicum and M. fortuitum known for infections in humans. This study shows that multiresistant RGM close to known human pathogens occur in grassland soils regardless the soil history of manuring.

  14. Evaluation of Various Culture Media for Detection of Rapidly Growing Mycobacteria from Patients with Cystic Fibrosis.

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    Preece, Clair L; Wichelhaus, Thomas A; Perry, Audrey; Jones, Amanda L; Cummings, Stephen P; Perry, John D; Hogardt, Michael

    2016-07-01

    Isolation of nontuberculous mycobacteria (NTM) from the sputum of patients with cystic fibrosis (CF) is challenging due to overgrowth by rapidly growing species that colonize the lungs of patients with CF. Extended incubation on Burkholderia cepacia selective agar (BCSA) has been recommended as an expedient culture method for the isolation of rapidly growing NTM in this setting. The aim of this study was to assess five selective media designed for the isolation of Burkholderia cepacia complex, along with two media designed for the isolation of mycobacteria (rapidly growing mycobacteria [RGM] medium and Middlebrook 7H11 agar), for their abilities to isolate NTM. All seven media were challenged with 147 isolates of rapidly growing mycobacteria and 185 isolates belonging to other species. RGM medium was then compared with the most selective brand of BCSA for the isolation of NTM from 224 sputum samples from patients with CF. Different agars designed for the isolation of B. cepacia complex varied considerably in their inhibition of other bacteria and fungi. RGM medium supported the growth of all isolates of mycobacteria and was more selective than any other medium. NTM were recovered from 17 of 224 sputum samples using RGM medium, compared with only 7 samples using the most selective brand of BCSA (P = 0.023). RGM medium offers a superior option, compared to other selective agars, for the isolation of rapidly growing mycobacteria from the sputum of patients with CF. Furthermore, the convenience of using RGM medium enables routine screening for rapidly growing NTM in all submitted sputum samples from patients with CF. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  15. measles immunisation growing peri-urban area of a mass a rapidly ...

    African Journals Online (AJOL)

    A mass measles immunisation campaign, with a target coverage rate of 85 - 90%, was launched in Khayelitsha, a rapidly growing urban township in the Cape Town area. Cross-sectional surveys of the measles immunisation status of resident 6 - 23-month-old infants were conducted immediately before, immediately after, ...

  16. Growing evidence for human health benefits of boron

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    Growing evidence from numerous laboratories using a variety of experimental models shows that boron is a bioactive beneficial, perhaps essential, element for humans. Reported beneficial actions of boron include arthritis alleviation or risk reduction; bone growth and maintenance; central nervous sys...

  17. In-vitro evaluation of the adhesion to polypropylene sutures of non-pigmented, rapidly growing mycobacteria.

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    Zamora, N; Esteban, J; Kinnari, T J; Celdrán, A; Granizo, J J; Zafra, C

    2007-09-01

    The ability of non-pigmented, rapidly growing mycobacteria (NPRGM) to attach to polypropylene sutures was evaluated using an in-vitro assay. Thirty clinical isolates and five culture collection strains of NPRGM, together with Staphylococcus epidermidis ATCC 35983, were tested. Mycobacterium fortuitum and Mycobacterium chelonae showed the highest attachment ability, which differed significantly from the results obtained with Mycobacterium peregrinum. According to these results, NPRGM are able to attach to polypropylene sutures, and the species implicated most frequently in human infection showed increased levels of attachment in comparison with the other mycobacteria studied.

  18. Mycobacterium aquiterrae sp. nov., a rapidly growing bacterium isolated from groundwater.

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    Lee, Jae-Chan; Whang, Kyung-Sook

    2017-10-01

    A strain representing a rapidly growing, Gram-stain-positive, aerobic, rod-shaped, non-motile, non-sporulating and non-pigmented species of the genus Mycobacterium, designated strain S-I-6T, was isolated from groundwater at Daejeon in Korea. The strain grew at temperatures between 10 and 37 °C (optimal growth at 25 °C), between pH 4.0 and 9.0 (optimal growth at pH 7.0) and at salinities of 0-5 % (w/v) NaCl, growing optimally with 2 % (w/v) NaCl. Phylogenetic analyses based on multilocus sequence analysis of the 16S rRNAgene, hsp65, rpoB and the 16S-23S internal transcribed spacer indicated that strain S-I-6T belonged to the rapidly growing mycobacteria, being most closely related to Mycobacterium sphagni. On the basis of polyphasic taxonomic analysis, the bacterial strain was distinguished from its phylogenetic neighbours by chemotaxonomic properties and other biochemical characteristics. DNA-DNA relatedness among strain S-I-6T and the closest phylogenetic neighbour strongly support the proposal that this strain represents a novel species within the genus Mycobacterium, for which the name Mycobacterium aquiterrae sp. nov. is proposed. The type strain is S-I-6T (=KACC 17600T=NBRC 109805T=NCAIM B 02535T).

  19. Antimicrobial susceptibility testing of rapidly growing mycobacteria by microdilution - Experience of a tertiary care centre

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    Set R

    2010-01-01

    Full Text Available Purpose: The objective of the study was to perform antimicrobial susceptibility testing of rapidly growing mycobacteria (RGM isolated from various clinically suspected cases of extrapulmonary tuberculosis, from January 2007 to April 2008, at a tertiary care centre in Mumbai. Materials and Methods: The specimens were processed for microscopy and culture using the standard procedures. Minimum inhibitory concentrations (MIC were determined by broth microdilution, using Sensititre CA MHBT. Susceptibility testing was also carried out on Mueller Hinton agar by the Kirby Bauer disc diffusion method. Results: Of the 1062 specimens received for mycobacterial cultures, 104 (9.79% grew mycobacteria. Of the mycobacterial isolates, six (5.76% were rapid growers. M. abscessus and M. chelonae appeared to be resistant organisms, with M. chelonae showing intermediate resistance to amikacin and minocycline. However, all the six isolates showed sensitivity to vancomycin and gentamicin by the disc diffusion test. Also all three isolates of M. abscessus were sensitive to piperacillin and erythromycin. Further studies are required to test their sensitivity to these four antimicrobials by using the microbroth dilution test, before they can be prescribed to patients. Conclusions: We wish to emphasize that reporting of rapidly growing mycobacteria from clinical settings, along with their sensitivity patterns, is an absolute need of the hour.

  20. Clinical management of rapidly growing mycobacterial cutaneous infections in patients after mesotherapy.

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    Regnier, Stéphanie; Cambau, Emmanuelle; Meningaud, Jean-Paul; Guihot, Amelie; Deforges, Lionel; Carbonne, Anne; Bricaire, François; Caumes, Eric

    2009-11-01

    Increasing numbers of patients are expressing an interest in mesotherapy as a method of reducing body fat. Cutaneous infections due to rapidly growing mycobacteria are a common complication of such procedures. We followed up patients who had developed cutaneous infections after undergoing mesotherapy during the period October 2006-January 2007. Sixteen patients were infected after mesotherapy injections performed by the same physician. All patients presented with painful, erythematous, draining subcutaneous nodules at the injection sites. All patients were treated with surgical drainage. Microbiological examination was performed on specimens that were obtained before and during the surgical procedure. Direct examination of skin smears demonstrated acid-fast bacilli in 25% of the specimens that were obtained before the procedure and 37% of the specimens obtained during the procedure; culture results were positive in 75% of the patients. Mycobacterium chelonae was identified in 11 patients, and Mycobacterium frederiksbergense was identified in 2 patients. Fourteen patients were treated with antibiotics, 6 received triple therapy as first-line treatment (tigecycline, tobramycin, and clarithromycin), and 8 received dual therapy (clarithromycin and ciprofloxacin). The mean duration of treatment was 14 weeks (range, 1-24 weeks). All of the patients except 1 were fully recovered 2 years after the onset of infection, with the mean time to healing estimated at 6.2 months (range, 1-15 months). This series of rapidly growing mycobacterial cutaneous infections highlights the difficulties in treating such infections and suggests that in vitro susceptibility to antibiotics does not accurately predict their clinical efficacy.

  1. Rapid urbanization and the growing threat of violence and conflict: a 21st century crisis.

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    Patel, Ronak B; Burkle, Frederick M

    2012-04-01

    As the global population is concentrated into complex environments, rapid urbanization increases the threat of conflict and insecurity. Many fast-growing cities create conditions of significant disparities in standards of living, which set up a natural environment for conflict over resources. As urban slums become a haven for criminal elements, youth gangs, and the arms trade, they also create insecurity for much of the population. Specific populations, such as women, migrants, and refugees, bear the brunt of this lack of security, with significant impacts on their livelihoods, health, and access to basic services. This lack of security and violence also has great costs to the general population, both economic and social. Cities have increasingly become the battlefield of recent conflicts as they serve as the seats of power and gateways to resources. International agencies, non-governmental organizations, and policy-makers must act to stem this tide of growing urban insecurity. Protecting urban populations and preventing future conflict will require better urban planning, investment in livelihood programs for youth, cooperation with local communities, enhanced policing, and strengthening the capacity of judicial systems.

  2. Cancer survivors in Switzerland: a rapidly growing population to care for

    Science.gov (United States)

    2013-01-01

    Background Cancer survivors are a heterogeneous group with complex health problems. Data concerning its total number and growing dynamics for Switzerland are scarce and outdated. Methods Population and mortality data were retrieved from the Swiss Federal Statistical Office (FSO). Incidence and relative survival for invasive cancers were computed using data from the cancer registries Geneva (1970–2009), St. Gallen - Appenzell (1980–2010), Grisons & Glarus (1989–2010), and Valais (1989–2010). We estimated prevalence for 1990–2010 using the Prevalence, Incidence Approach MODel (PIAMOD) method. We calculated trends in prevalence estimates by Joinpoint analysis. Projections were extrapolated using the above models and based on time trends of the period 2007–2010. Results The estimated number of cancer survivors increased from 139′717 in 1990 (2.08% of the population) to 289′797 persons in 2010 (3.70%). The growth rate shows an exponential shape and was 3.3% per year in the period 2008 to 2010. Almost half of the survivors have a history of breast, prostate or colorectal cancer. Among cancer survivors, 55% are women but the increases have been more marked in men (p Switzerland. Conclusions There is a rapidly growing population of cancer survivors in Switzerland whose needs and concerns are largely unknown. PMID:23764068

  3. Rapidly growing tropical trees mobilize remarkable amounts of nitrogen, in ways that differ surprisingly among species.

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    Russell, Ann E; Raich, James W

    2012-06-26

    Fast-growing forests such as tropical secondary forests can accumulate large amounts of carbon (C), and thereby play an important role in the atmospheric CO(2) balance. Because nitrogen (N) cycling is inextricably linked with C cycling, the question becomes: Where does the N come from to match high rates of C accumulation? In unique experimental 16-y-old plantations established in abandoned pasture in lowland Costa Rica, we used a mass-balance approach to quantify N accumulation in vegetation, identify sources of N, and evaluate differences among tree species in N cycling. The replicated design contained four broad-leaved evergreen tree species growing under similar environmental conditions. Nitrogen uptake was rapid, reaching 409 (± 30) kg · ha(-1) · y(-1), double the rate reported from a Puerto Rican forest and greater than four times that observed at Hubbard Brook Forest (New Hampshire, USA). Nitrogen amassed in vegetation was 874 (± 176) kg · ha(-1), whereas net losses of soil N (0-100 cm) varied from 217 (±146) to 3,354 (± 915) kg · ha(-1) (P = 0.018) over 16 y. Soil C:N, δ(13)C values, and N budgets indicated that soil was the main source of biomass N. In Vochysia guatemalensis, however, N fixation contributed >60 kg · ha(-1) · y(-1). All species apparently promoted soil N turnover, such that the soil N mean residence time was 32-54 y, an order of magnitude lower than the global mean. High rates of N uptake were associated with substantial N losses in three of the species, in which an average of 1.6 g N was lost for every gram of N accumulated in biomass.

  4. The spatial biology of transcription and translation in rapidly growing Escherichia coli

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    Somenath eBakshi

    2015-07-01

    Full Text Available Single-molecule fluorescence provides high resolution spatial distributions of ribosomes and RNA polymerase (RNAP in live, rapidly growing E. coli. Ribosomes are more strongly segregated from the nucleoids (chromosomal DNA than previous widefield fluorescence studies suggested. While most transcription may be co-translational, the evidence indicates that most translation occurs on free mRNA copies that have diffused from the nucleoids to a ribosome-rich region. Analysis of time-resolved images of the nucleoid spatial distribution after treatment with the transcription-halting drug rifampicin and the translation-halting drug chloramphenicol shows that both drugs cause nucleoid contraction on the 0-3 min timescale. This is consistent with the transertion hypothesis. We suggest that the longer-term (20-30 min nucleoid expansion after Rif treatment arises from conversion of 70S-polysomes to 30S and 50S subunits, which readily penetrate the nucleoids. Monte Carlo simulations of a polymer bead model built to mimic the chromosomal DNA and ribosomes (either 70S-polysomes or 30S and 50S subunits explain spatial segregation or mixing of ribosomes and nucleoids in terms of excluded volume and entropic effects alone. A comprehensive model of the transcription-translation-transertion system incorporates this new information about the spatial organization of the E. coli cytoplasm. We propose that transertion, which radially expands the nucleoids, is essential for recycling of 30S and 50S subunits from ribosome-rich regions back into the nucleoids. There they initiate co-transcriptional translation, which is an important mechanism for maintaining RNAP forward progress and protecting the nascent mRNA chain. Segregation of 70S-polysomes from the nucleoid may facilitate rapid growth by shortening the search time for ribosomes to find free mRNA concentrated outside the nucleoid and the search time for RNAP concentrated within the nucleoid to find transcription

  5. An automated system for rapid non-destructive enumeration of growing microbes.

    Directory of Open Access Journals (Sweden)

    Roanna London

    Full Text Available BACKGROUND: The power and simplicity of visual colony counting have made it the mainstay of microbiological analysis for more than 130 years. A disadvantage of the method is the long time required to generate visible colonies from cells in a sample. New rapid testing technologies generally have failed to maintain one or more of the major advantages of culture-based methods. PRINCIPAL FINDINGS: We present a new technology and platform that uses digital imaging of cellular autofluorescence to detect and enumerate growing microcolonies many generations before they become visible to the eye. The data presented demonstrate that the method preserves the viability of the microcolonies it detects, thus enabling generation of pure cultures for microbial identification. While visual colony counting detects Escherichia coli colonies containing about 5x10(6 cells, the new imaging method detects E. coli microcolonies when they contain about 120 cells and microcolonies of the yeast Candida albicans when they contain only about 12 cells. We demonstrate that digital imaging of microcolony autofluorescence detects a broad spectrum of prokaryotic and eukaryotic microbes and present a model for predicting the time to detection for individual strains. Results from the analysis of environmental samples from pharmaceutical manufacturing plants containing a mixture of unidentified microbes demonstrate the method's improved test turnaround times. CONCLUSION: This work demonstrates a new technology and automated platform that substantially shortens test times while maintaining key advantages of the current methods.

  6. Mycobacterium celeriflavum sp. nov., a rapidly growing scotochromogenic bacterium isolated from clinical specimens.

    Science.gov (United States)

    Shahraki, Abdolrazagh Hashemi; Çavuşoğlu, Cengiz; Borroni, Emanuele; Heidarieh, Parvin; Koksalan, Orhan Kaya; Cabibbe, Andrea Maurizio; Hashemzadeh, Mohamad; Mariottini, Alessandro; Mostafavi, Ehsan; Cittaro, Davide; Feizabadi, Mohamad Mehdi; Lazarevic, Dejan; Yaghmaei, Farhad; Molinari, Gian Lorenzo; Camaggi, Anna; Tortoli, Enrico

    2015-02-01

    Six strains of a rapidly growing scotochromogenic mycobacterium were isolated from pulmonary specimens of independent patients. Biochemical and cultural tests were not suitable for their identification. The mycolic acid pattern analysed by HPLC was different from that of any other mycobacterium. Genotypic characterization, targeting seven housekeeping genes, revealed the presence of microheterogeneity in all of them. Different species were more closely related to the test strains in various regions: the type strain of Mycobacterium moriokaense showed 99.0 % 16S rRNA gene sequence similarity, and 91.5-96.5 % similarity for the remaining six regions. The whole genome sequences of the proposed type strain and that of M. moriokaense presented an average nucleotide identity (ANI) of 82.9 %. Phylogenetic analysis produced poorly robust trees in most genes with the exception of rpoB and sodA where Mycobacterium flavescens and Mycobacterium novocastrense were the closest species. This phylogenetic relatedness was confirmed by the tree inferred from five concatenated genes, which was very robust. The polyphasic characterization of the test strains, supported by the ANI value, demonstrates that they belong to a previously unreported species, for which the name Mycobacterium celeriflavum sp. nov. is proposed. The type strain is AFPC-000207(T) ( = DSM 46765(T) = JCM 18439(T)). © 2015 IUMS.

  7. Isolation of Rapidly Growing Nontuberculous Mycobacteria in Wounds Following Combat-Related Injury.

    Science.gov (United States)

    Fiske, Lauren C; Homeyer, Diane C; Zapor, Michael; Hartzell, Joshua; Warkentien, Tyler; Weintrob, Amy C; Ganesan, Anuradha; Burgess, Timothy; Snesrud, Erik; Waterman, Paige; Nielsen, Lindsey; Ressner, Roseanne A

    2016-06-01

    Rapidly growing nontuberculous mycobacteria (RGNTM) have yet to be described in combat-related injuries. This study investigates the epidemiology, clinical findings, treatment, and outcomes of RGNTM infections among combat casualties wounded in Afghanistan from 2010 to 2012. Patients with RGNTM were identified from the Department of Defense Trauma Registry through the Trauma Infectious Disease Outcomes Study. Trauma history, surgical management, and clinical data were collected. Six isolates from patients requiring antimycobacterial therapy were sequenced. Seventeen cases were identified. Six cases, predominantly associated with Mycobacterium abscessus, required aggressive debridement and a median of 180 days of multidrug antimycobacterial therapy that included clofazimine. M. abscessus isolates expressed the erythromycin resistance methylase (erm(41)) gene for inducible macrolide resistance, yet there were no clinical treatment failures when macrolides were utilized in combination therapy. No clonal similarity between M. abscessus isolates was found. Eleven cases had positive wound cultures, but did not require antimycobacterial therapy. The median duration of time of injury to first detection of a RGNTM was 57 days. This represents the first report of RGNTM infections in war-wounded patients. RGNTM should be recognized as potential pathogens in grossly infected combat wounds. Surgical debridement and multidrug antimycobacterial therapy, when clinically indicated, was associated with satisfactory clinical outcomes. Reprint & Copyright © 2016 Association of Military Surgeons of the U.S.

  8. Nosocomial rapidly growing mycobacterial infections following laparoscopic surgery: CT imaging findings

    Energy Technology Data Exchange (ETDEWEB)

    Volpato, Richard [Cassiano Antonio de Moraes University Hospital, Department of Diagnostic Radiology, Vitoria, ES (Brazil); Campi de Castro, Claudio [University of Sao Paulo Medical School, Department of Radiology, Cerqueira Cesar, Sao Paulo (Brazil); Hadad, David Jamil [Cassiano Antonio de Moraes University Hospital, Nucleo de Doencas Infecciosas, Department of Internal Medicine, Vitoria, ES (Brazil); Silva Souza Ribeiro, Flavya da [Laboratorio de Patologia PAT, Department of Diagnostic Radiology, Unit 1473, Vitoria, ES (Brazil); Filho, Ezequiel Leal [UNIMED Diagnostico, Department of Diagnostic Radiology, Unit 1473, Vitoria, ES (Brazil); Marcal, Leonardo P. [The University of Texas M D Anderson Cancer Center, Department of Diagnostic Radiology, Unit 1473, Houston, TX (United States)

    2015-09-15

    To identify the distribution and frequency of computed tomography (CT) findings in patients with nosocomial rapidly growing mycobacterial (RGM) infection after laparoscopic surgery. A descriptive retrospective study in patients with RGM infection after laparoscopic surgery who underwent CT imaging prior to initiation of therapy. The images were analyzed by two radiologists in consensus, who evaluated the skin/subcutaneous tissues, the abdominal wall, and intraperitoneal region separately. The patterns of involvement were tabulated as: densification, collections, nodules (≥1.0 cm), small nodules (<1.0 cm), pseudocavitated nodules, and small pseudocavitated nodules. Twenty-six patients met the established criteria. The subcutaneous findings were: densification (88.5 %), small nodules (61.5 %), small pseudocavitated nodules (23.1 %), nodules (38.5 %), pseudocavitated nodules (15.4 %), and collections (26.9 %). The findings in the abdominal wall were: densification (61.5 %), pseudocavitated nodules (3.8 %), and collections (15.4 %). The intraperitoneal findings were: densification (46.1 %), small nodules (42.3 %), nodules (15.4 %), and collections (11.5 %). Subcutaneous CT findings in descending order of frequency were: densification, small nodules, nodules, small pseudocavitated nodules, pseudocavitated nodules, and collections. The musculo-fascial plane CT findings were: densification, collections, and pseudocavitated nodules. The intraperitoneal CT findings were: densification, small nodules, nodules, and collections. (orig.)

  9. An assessment of the maxilla after rapid maxillary expansion using cone beam computed tomography in growing children.

    Science.gov (United States)

    Woller, Jessica L; Kim, Ki Beom; Behrents, Rolf G; Buschang, Peter H

    2014-01-01

    With the advent of cone beam computed tomography (CBCT), it is now possible to quantitatively evaluate the effects of rapid maxillary expansion (RME) on the entire maxillary complex in growing patients. The purpose of this study is to use three-dimensional images to evaluate the displacement that occurs at the circummaxillary sutures (frontonasal, zygomaticomaxillary, intermaxillary, midpalatal, and transpalatal sutures) following rapid maxillary expansion in growing children. The CBCT scans of 25 consecutively treated RME patients (10 male, 15 female) with mean age of 12.3±2.6 years, were examined before expansion and immediately following the last activation of the expansion appliance. Statistically significant (Pmaxillary first molars due to RME was also statistically significant. There was no statistically significant displacement of the transpalatal suture. Rapid maxillary expansion results in significant displacement of the bones of circummaxillary sutures in growing children.

  10. Two novel species of rapidly growing mycobacteria: Mycobacterium lehmannii sp. nov. and Mycobacterium neumannii sp. nov.

    Science.gov (United States)

    Nouioui, Imen; Sangal, Vartul; Carro, Lorena; Teramoto, Kanae; Jando, Marlen; Montero-Calasanz, Maria Del Carmen; Igual, José Mariano; Sutcliffe, Iain; Goodfellow, Michael; Klenk, Hans-Peter

    2017-12-01

    Two rapidly growing mycobacteria with identical 16S rRNA gene sequences were the subject of a polyphasic taxonomic study. The strains formed a well-supported subclade in the mycobacterial 16S rRNA gene tree and were most closely associated with the type strain of Mycobacterium novocastrense. Single and multilocus sequence analyses based on hsp65, rpoB and 16S rRNA gene sequences showed that strains SN 1900T and SN 1904T are phylogenetically distinct but share several chemotaxonomic and phenotypic features that are are consistent with their classification in the genus Mycobacterium. The two strains were distinguished by their different fatty acid and mycolic acid profiles, and by a combination of phenotypic features. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values for strains SN 1900T and SN 1904T were 61.0 % and 94.7 %, respectively; in turn, the corresponding dDDH and ANI values with M. novocastrense DSM 44203T were 41.4 % and 42.8 % and 89.3 % and 89.5 %, respectively. These results show that strains SN1900T and SN 1904T form new centres of taxonomic variation within the genus Mycobacterium. Consequently, strains SN 1900T (40T=CECT 8763T=DSM 43219T) and SN 1904T (2409T=CECT 8766T=DSM 43532T) are considered to represent novel species, for which the names Mycobacteriumlehmannii sp. nov. and Mycobacteriumneumannii sp. nov. are proposed. A strain designated as 'Mycobacteriumacapulsensis' was shown to be a bona fide member of the putative novel species, M. lehmannii.

  11. To grow or not to grow: hair morphogenesis and human genetic hair disorders.

    Science.gov (United States)

    Duverger, Olivier; Morasso, Maria I

    2014-01-01

    Mouse models have greatly helped in elucidating the molecular mechanisms involved in hair formation and regeneration. Recent publications have reviewed the genes involved in mouse hair development based on the phenotype of transgenic, knockout and mutant animal models. While much of this information has been instrumental in determining molecular aspects of human hair development and cycling, mice exhibit a specific pattern of hair morphogenesis and hair distribution throughout the body that cannot be directly correlated to human hair. In this mini-review, we discuss specific aspects of human hair follicle development and present an up-to-date summary of human genetic disorders associated with abnormalities in hair follicle morphogenesis, structure or regeneration. Published by Elsevier Ltd.

  12. An assessment of the maxilla after rapid maxillary expansion using cone beam computed tomography in growing children

    OpenAIRE

    Woller,Jessica L.; Ki Beom Kim; Behrents, Rolf G.; Buschang, Peter H.

    2014-01-01

    INTRODUCTION: With the advent of cone beam computed tomography (CBCT), it is now possible to quantitatively evaluate the effects of rapid maxillary expansion (RME) on the entire maxillary complex in growing patients. OBJECTIVE: The purpose of this study is to use three-dimensional images to evaluate the displacement that occurs at the circummaxillary sutures (frontonasal, zygomaticomaxillary, intermaxillary, midpalatal, and transpalatal sutures) following rapid maxillary expansion in gro...

  13. Mycobacterium iranicum sp. nov., a rapidly growing scotochromogenic species isolated from clinical specimens on three different continents

    NARCIS (Netherlands)

    Shojaei, H.; Daley, C.; Gitti, Z.; Hashemi, A.; Heidarieh, P.; Moore, E.R.; Naser, A.D.; Russo, C.; Ingen, J. van; Tortoli, E.

    2013-01-01

    The isolation and characterization of a novel, rapidly growing, scotochromogenic mycobacterial species is reported. Eight independent strains were isolated from clinical specimens from six different countries of the world, two in Iran, two in Italy and one in each of following countries: Greece, The

  14. Mandibular response after rapid maxillary expansion in class II growing patients: a pilot randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Roberta Lione

    2017-11-01

    Full Text Available Abstract Background The aim of this pilot randomized controlled trial (RCT was to evaluate the sagittal mandibular response induced by rapid maxillary expansion (RME therapy in mixed dentition patients with class II malocclusion, comparing the effects of bonded RME and banded RME with a matched untreated class II control group. Methods This RCT was designed in parallel with an allocation ratio of 1:1:1. The sample consisted of 30 children with a mean age of 8.1 ± 0.6 years who were randomly assigned to three groups: group 1 treated with bonded RME, group 2 treated with banded RME, and group 3 the untreated control group. All patients met the following inclusion criteria: early mixed dentition, class II molar relationship, transverse discrepancy ≥ 4 mm, overjet ≥ 5 mm, and prepubertal skeletal maturity stage (CS1–CS2. The expansion screw was activated one quarter of a turn per day (0.25 mm until overcorrection was reached. For each subject, lateral cephalograms and plaster casts were obtained before treatment (T1 and after 1 year (T2. A randomization list was created for the group assignment, with an allocation ratio of 1:1:1. The observer who performed all the measurements was blinded to group assignment. The study was single-blinded in regard to statistical analysis. Results RME was effective in the correction of maxillary deficiency. Class II patients treated with both types of RME showed no significant improvement of the anteroposterior relationship of the maxilla and the mandible at both skeletal and occlusal levels. The acrylic splint RME had significant effects on reducing the skeletal vertical dimension and the gonial angle. Conclusions The orthopedic expansion did not affect the sagittal relationship of class II patients treated in the early mixed dentition when compared with the untreated control group. Additional studies with a larger sample are warranted to elucidate individual variations in dento-skeletal mandibular

  15. Mandibular response after rapid maxillary expansion in class II growing patients: a pilot randomized controlled trial.

    Science.gov (United States)

    Lione, Roberta; Brunelli, Valerio; Franchi, Lorenzo; Pavoni, Chiara; Quiroga Souki, Bernardo; Cozza, Paola

    2017-11-06

    The aim of this pilot randomized controlled trial (RCT) was to evaluate the sagittal mandibular response induced by rapid maxillary expansion (RME) therapy in mixed dentition patients with class II malocclusion, comparing the effects of bonded RME and banded RME with a matched untreated class II control group. This RCT was designed in parallel with an allocation ratio of 1:1:1. The sample consisted of 30 children with a mean age of 8.1 ± 0.6 years who were randomly assigned to three groups: group 1 treated with bonded RME, group 2 treated with banded RME, and group 3 the untreated control group. All patients met the following inclusion criteria: early mixed dentition, class II molar relationship, transverse discrepancy ≥ 4 mm, overjet ≥ 5 mm, and prepubertal skeletal maturity stage (CS1-CS2). The expansion screw was activated one quarter of a turn per day (0.25 mm) until overcorrection was reached. For each subject, lateral cephalograms and plaster casts were obtained before treatment (T1) and after 1 year (T2). A randomization list was created for the group assignment, with an allocation ratio of 1:1:1. The observer who performed all the measurements was blinded to group assignment. The study was single-blinded in regard to statistical analysis. RME was effective in the correction of maxillary deficiency. Class II patients treated with both types of RME showed no significant improvement of the anteroposterior relationship of the maxilla and the mandible at both skeletal and occlusal levels. The acrylic splint RME had significant effects on reducing the skeletal vertical dimension and the gonial angle. The orthopedic expansion did not affect the sagittal relationship of class II patients treated in the early mixed dentition when compared with the untreated control group. Additional studies with a larger sample are warranted to elucidate individual variations in dento-skeletal mandibular response to the maxillary expansion protocol in class-II-growing

  16. [The diagnosis and treatment of rapidly growing non-tuberculous mycobacterial keratitis].

    Science.gov (United States)

    Guan, Huai-Jin; Cheng, Zheng-Ping; Yin, Li; Wu, Yu-Yu; Hu, Nan; Zhang, Jun-Fang; Shi, Hai-Hong

    2009-06-01

    To study the clinical features, diagnosis and treatment of non-tuberculous mycobacterial keratitis (NTMK). It was retrospective case series study. Twelve eyes in 12 patients with NTMK following corneal foreign body trauma in 2007 were studied retrospectively including the case histories, clinical findings, laboratory examinations, diagnosis, treatment and prognosis. The main laboratory examination included corneal scrapings by culturing, polymerase chain reaction (PCR) and transmission electron microscopy (TEM), corneal lesions by histopathologic examinations and TEM. The patients received local and systemic antibiotics therapy, lesion cleaning followed by cauterization with tincture of iodine (5%) and (or) keratoplasty. All cases had a history of corneal trauma, there was corneal metallic foreign body removal at one hospital in 11 cases, corneal reed trauma in 1 case. The characteristic signs involved grayish-blue crystalloid keratopathy, multifocal infiltrates, satellites, radical form changes in the Descemet's membrane. The results of laboratory examinations of the scrapings of the cornea infection were as follows: all cultures (12/12) were positive for rapidly growing mycobacteria, and isolates from 5 patients were all diagnosed as mycobacterium chelonae subspecies abscess; acid-fast staining revealed positive bacilli in all the 4 patients; seven of 8 patients were positive for bacterium by PCR. Transmission electron microscopy in all the 3 specimens showed many slender rod-shaped or short coarse-shaped bacteria which were phagocytized by monocytes, and some necrotic tissue. Infections in 10 eyes were resolved by combined treatment regimen including a combination of antimicrobial agents (amikacin, rifampin, gatifloxacin, ciprofloxacin, azithromycin and/or ofloxacin, etc.) and local lesion cleaning followed by cauterization with 5% tincture of iodine within 2-5 months; two cases resolved by keratoplasty which poorly responded to antibiotic therapy for 6 months

  17. To grow or not to grow: Hair morphogenesis and human genetic hair disorders

    OpenAIRE

    Duverger, Olivier; Morasso, Maria I.

    2013-01-01

    Mouse models have greatly helped in elucidating the molecular mechanisms involved in hair formation and regeneration. Recent publications have reviewed the genes involved in mouse hair development based on the phenotype of transgenic, knockout and mutant animal models. While much of this information has been instrumental in determining molecular aspects of human hair development and cycling, mice exhibit a specific pattern of hair morphogenesis and hair distribution throughout the body that c...

  18. New quantitative patterns of the growing trachea in human fetuses.

    Science.gov (United States)

    Szpinda, Michał; Daroszewski, Marcin; Szpinda, Anna; Woźniak, Alina; Wiśniewski, Marcin; Mila-Kierzenkowska, Celestyna; Baumgart, Mariusz; Paruszewska-Achtel, Monika

    2012-06-01

    Rapid progress in perinatal medicine has resulted in numerous tracheo-bronchial interventions on fetal and neonatal airways. The present study was performed to compile normative data for tracheal dimensions at varying gestational ages. Using anatomical dissection, digital image analysis (NIS-Elements BR 3.0) and statistical analysis (Wilcoxon signed-rank test, Student's t test, one-way ANOVA, post-hoc Bonferroni test, linear and nonlinear regression analysis) a range of the 4 variables (length in mm, middle external transverse diameter in mm, proximal internal cross-sectional area in mm², internal volume in mm³) for the trachea in 73 spontaneously aborted human fetuses (39 male, 34 female) aged 14-25 weeks was examined. No significant male-female differences were found (P>0.05). The length ranged from 10.37±2.15 to 26.54±0.26 mm as y=-65.098 + 28.796 × ln (Age) ±1.794 (R²=0.82). The middle external transverse diameter varied from 2.53±0.09 to 5.09±0.42 mm with the model y=-11.020 + 5.049 × ln (Age) ±0.330 (R²=0.81). The trachea indicated a proportional evolution because the middle external transverse diameter-to-length ratio was stable (0.23±0.03). The proximal internal cross-sectional area rose from 1.46±0.04 to 5.76±1.04 mm² as y=-3.562 + 0.352 × Age ±0.519 (R²=0.76). The internal volumetric growth from 11.89±2.49 to 119.63±4.95 mm³ generated the function y=-135.248 + 9.919 × Age ±10.478 (R²=0.86). The growth in both length and middle external transverse diameter of the trachea follows logarithmic functions, whereas growth of both its proximal internal cross-sectional area and internal volume follow linear functions. The length and middle external transverse diameter of the trachea develop proportionally to each other. The tracheal dimensions may be helpful in the prenatal diagnosis and monitoring of tracheal malformations and obstructive anomalies of the upper respiratory tract.

  19. 78 FR 24692 - Standards for the Growing, Harvesting, Packing, and Holding of Produce for Human Consumption...

    Science.gov (United States)

    2013-04-26

    ... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption; Extension of Comment Periods..., Harvesting, Packing, and Holding of Produce for Human Consumption'' that appeared in the Federal Register of... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption'' with a 120-day comment period...

  20. Rapid Prototyping and the Human Factors Engineering Process

    Science.gov (United States)

    2016-08-29

    Rapid prototyping and the human factors • • engineering process David Beevis* and Gaetan St Denist *Senior Human Factors Engineer, Defence and...qr-..2. 9 Rapid prototyping or ’virtual prototyping ’ of human-machine interfaces offers the possibility of putting the human operator ’in the loop...8217 without the effort and cost associated with conventional man-in-the-loop simulation. Advocates suggest that rapid prototyping is compatible with

  1. Multistate US Outbreak of Rapidly Growing Mycobacterial Infections Associated with Medical Tourism to the Dominican Republic, 2013-2014(1).

    Science.gov (United States)

    Schnabel, David; Esposito, Douglas H; Gaines, Joanna; Ridpath, Alison; Barry, M Anita; Feldman, Katherine A; Mullins, Jocelyn; Burns, Rachel; Ahmad, Nina; Nyangoma, Edith N; Nguyen, Duc B; Perz, Joseph F; Moulton-Meissner, Heather A; Jensen, Bette J; Lin, Ying; Posivak-Khouly, Leah; Jani, Nisha; Morgan, Oliver W; Brunette, Gary W; Pritchard, P Scott; Greenbaum, Adena H; Rhee, Susan M; Blythe, David; Sotir, Mark

    2016-08-01

    During 2013, the Maryland Department of Health and Mental Hygiene in Baltimore, MD, USA, received report of 2 Maryland residents whose surgical sites were infected with rapidly growing mycobacteria after cosmetic procedures at a clinic (clinic A) in the Dominican Republic. A multistate investigation was initiated; a probable case was defined as a surgical site infection unresponsive to therapy in a patient who had undergone cosmetic surgery in the Dominican Republic. We identified 21 case-patients in 6 states who had surgery in 1 of 5 Dominican Republic clinics; 13 (62%) had surgery at clinic A. Isolates from 12 (92%) of those patients were culture-positive for Mycobacterium abscessus complex. Of 9 clinic A case-patients with available data, all required therapeutic surgical intervention, 8 (92%) were hospitalized, and 7 (78%) required ≥3 months of antibacterial drug therapy. Healthcare providers should consider infection with rapidly growing mycobacteria in patients who have surgical site infections unresponsive to standard treatment.

  2. Rapidly growing Mycobacterium infections after cosmetic surgery in medical tourists: the Bronx experience and a review of the literature

    OpenAIRE

    Lucas R. Cusumano; Vivy Tran; Aileen Tlamsa; Philip Chung; Robert Grossberg; Gregory Weston; Uzma N. Sarwar

    2017-01-01

    Background: Medical tourism is increasingly popular for elective cosmetic surgical procedures. However, medical tourism has been accompanied by reports of post-surgical infections due to rapidly growing mycobacteria (RGM). The authors’ experience working with patients with RGM infections who have returned to the USA after traveling abroad for cosmetic surgical procedures is described here. Methods: Patients who developed RGM infections after undergoing cosmetic surgeries abroad and who pre...

  3. A novel culture medium for isolation of rapidly-growing mycobacteria from the sputum of patients with cystic fibrosis.

    Science.gov (United States)

    Preece, Clair L; Perry, Audrey; Gray, Bethany; Kenna, Dervla T; Jones, Amanda L; Cummings, Stephen P; Robb, Ali; Thomas, Matthew F; Brodlie, Malcolm; O'Brien, Christopher J; Bourke, Stephen J; Perry, John D

    2016-03-01

    Isolation of mycobacteria from the sputum of patients with cystic fibrosis (CF) is challenging due to the overgrowth of cultures by other bacteria and fungi. In this setting, Burkholderia cepacia selective agar (BCSA) has been recommended as a convenient and effective culture medium for the isolation of rapidly-growing, non-tuberculous mycobacteria (NTM). A novel selective culture medium (RGM medium) was evaluated for the isolation of rapidly-growing NTM from the sputum of children and adults with CF. A total of 118 isolates of rapidly-growing mycobacteria and 98 other bacteria and fungi were inoculated onto RGM medium. These were assessed for growth at 30°C over a seven day period. A total of 502 consecutive sputum samples were collected from 210 patients with CF. Each sample was homogenized and cultured onto RGM medium and also onto BCSA. Cultures were incubated for 10days at 30°C. Of 118 isolates of mycobacteria all but one grew well on RGM medium, whereas 94% of other bacteria and fungi were inhibited. A total of 55 sputum samples (from 33 distinct patients) yielded NTM using a combination of both RGM and BCSA (prevalence: 15.7%). NTM were recovered from 54 sputum samples using RGM medium compared with only 17 samples using BCSA (sensitivity 98% vs. 31%; P≤0.0001). A total of 419 isolates of non-mycobacteria were recovered from sputum samples on BCSA compared with 46 on RGM medium. RGM medium offers a simple and effective culture method for the isolation of rapidly-growing mycobacteria from sputum samples from patients with CF without decontamination of samples. RGM medium allows for the systematic screening of all sputum samples routinely referred for culture from patients with CF. Copyright © 2015 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

  4. An assessment of the maxilla after rapid maxillary expansion using cone beam computed tomography in growing children

    Directory of Open Access Journals (Sweden)

    Jessica L. Woller

    2014-01-01

    Full Text Available INTRODUCTION: With the advent of cone beam computed tomography (CBCT, it is now possible to quantitatively evaluate the effects of rapid maxillary expansion (RME on the entire maxillary complex in growing patients. OBJECTIVE: The purpose of this study is to use three-dimensional images to evaluate the displacement that occurs at the circummaxillary sutures (frontonasal, zygomaticomaxillary, intermaxillary, midpalatal, and transpalatal sutures following rapid maxillary expansion in growing children. METHODS: The CBCT scans of 25 consecutively treated RME patients (10 male, 15 female with mean age of 12.3 ± 2.6 years, were examined before expansion and immediately following the last activation of the expansion appliance. RESULTS: Statistically significant (P < 0.05 amounts of separation were found for the displacement of the bones of the frontonasal suture, the intermaxillary suture, the zygomaticomaxillary sutures, and the midpalatal suture. The change in angulation of the maxillary first molars due to RME was also statistically significant. There was no statistically significant displacement of the transpalatal suture. CONCLUSIONS: Rapid maxillary expansion results in significant displacement of the bones of circummaxillary sutures in growing children.

  5. The impact of entrepreneurial capital and rapidly growing firms: the Canadian example

    DEFF Research Database (Denmark)

    Keen, Christian; Etemad, Hamid

    2011-01-01

    World-class competitiveness is no longer an option for firms seeking growth and survival in the increasingly competitive, dynamic and interconnected world. This paper expands on the concept of entrepreneurial capital and formalizes it as a catalyst that augments other productive factors. It provi...... hostile environments that also suffer from poor resources, this research offers significant lessons with implications for emerging firms, industries and associated regions that aspire to grow faster...

  6. Touch five factors to growing and leading a human organization

    CERN Document Server

    Maffin, Tod

    2014-01-01

    For better or worse, digital business has fundamentally changed how organizations hire, market their services, and connect with stakeholders. The problem is, in an effort to use technology to connect more effectively, we have lost the humanity - that critical person-to-person connection. This book will show you how to restore that connection.

  7. Measurement of human embryonic stem cell in the growing cycle

    Science.gov (United States)

    Li, X.; Zhao, L.; Oh, Steve K. W.; Chong, W. K.; Ong, J. K.; Chen, Allen K.; Choo, Andre B. H.

    2008-09-01

    A measurement and imaging system has been developed for in-line continuous measurement of live, unmodified, human embryonic stem cells (hESC). The measurement will not affect cell growth, structure, sterility and suitability for clinical use. The stem cell imaging system (SCIS) can be used to support the optimization of automated stem cell growth for invitro study and for high-volume bio-manufacture. This paper present the experimental and analysis for the optimization of system parameters. A non-linear lighting is developed to obtain a clear images. The individual cluster can be traced from day one to day two. The whole system is calibrated with measurement microscope and haemacytometer. The measurement accuracy is better than 90%.

  8. Rapidly growing cystic vestibular schwannoma with sudden onset facial palsy, ten years after subtotal excision.

    Science.gov (United States)

    Dandinarasaiah, Manjunath; Grinblat, Golda; Prasad, Sampath Chandra; Taibah, Abdelkader; Sanna, Mario

    2017-07-19

    An elderly male patient diagnosed with a right-sided cystic vestibular schwannoma (CVS) at our center underwent a translabyrinthine approach with a subtotal excision to preserve the facial nerve (FN). The tumor grew slowly for the first 9 years but in the subsequent 2 years grew rapidly, with the patient developing a FN paralysis. Using the previous approach, a second surgery was done and the tumor was excised, leaving behind a sheath of tumor on the facial and lower cranial nerves. This case demonstrates that CVSs show unpredictable growth patterns and need to be followed up for a longer period of time. Laryngoscope, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  9. A rapidly growing moraine-dammed glacial lake on Ngozumpa Glacier, Nepal

    Science.gov (United States)

    Thompson, Sarah S.; Benn, Douglas I.; Dennis, Kathryn; Luckman, Adrian

    2012-04-01

    Moraine-dammed glacial lakes are becoming increasingly common in the Himalaya as a result of glacier mass loss, causing concern about glacier lake outburst flood risk. In addition to extant lakes, the potential exists for many more to form, as more glaciers ablate down to the level of potential moraine dams. In this paper, we document the recent rapid growth of, a moraine-dammed lake on Ngozumpa Glacier, Nepal. Using a combination of ground-based mapping and sonar surveys, aerial photographs (< 1 m resolution), and ASTER imagery (15 m resolution), processes and rates of lake expansion have been determined. The lake first formed between 1984 and 1992 when collapse of an englacial conduit allowed water to accumulate at the level of a gap in the lateral moraine, ~km from the glacier terminus. Lake growth was initially slow, but since 2001 it has undergone exponential growth at an average rate of 10% y-1. In 2009, the lake area was 300,000 m2, and its volume was at least 2.2 million m3. Calving, subaqueous melting, and melting of subaerial ice faces all contribute to the expansion of the lake; but large-scale, full-height slab calving is now the dominant contributor to growth. Comparison with other lakes in the region indicate that lake growth will likely continue unchecked whilst the spillway remains at its current level and may attain a volume of hundreds of millions of cubic metres within the next few decades.

  10. Neonatal sepsis in a rapidly growing, tertiary neonatal intensive care unit: Trends over 18 years.

    Science.gov (United States)

    Heo, Ju Sun; Shin, Seung Han; Jung, Young Hwa; Kim, Ee-Kyung; Choi, Eun Hwa; Kim, Han-Suk; Lee, Hoan Jong; Choi, Jung-Hwan

    2015-10-01

    We investigated changes in the admission patterns of neonatal intensive care units and the epidemiology of neonatal sepsis following the rapid expansion and improvements in neonatal intensive care. Data on the admission of neonates with culture-proven sepsis between 1996 and 2013 (period I, 1996-2005; period II, 2006-2013) were collected retrospectively. The admission of extremely low-birthweight (ELBW) infants increased between periods I and II (11.1 vs 28.7 infants per 1000 live births, P sepsis among all infants and ELBW infants increased (all infants, 5.9 vs 12.7 cases per 1000 live births; ELBW infants, 189.5 vs 290.1 cases per 1000 live births). In ELBW infants, the incidence of sepsis caused by coagulase-negative Staphylococcus (CONS), significantly increased during period II (8.8 vs 25.4%, P = 0.039). On multivariate analysis, central vascular catheters and prolonged hospitalization were independently associated with increased sepsis rate, particularly CONS in ELBW infants. The inborn admission rate for ELBW infants has increased significantly and is accompanied by improved survival and longer hospital stay. The incidence of neonatal sepsis, particularly in ELBW infants, has also increased, and CONS has emerged as a major pathogen. Central vascular catheters and prolonged hospitalization could be independent risk factors for the increased sepsis rate, particularly sepsis due to CONS. © 2015 Japan Pediatric Society.

  11. 78 FR 69605 - Standards for the Growing, Harvesting, Packing, and Holding of Produce for Human Consumption...

    Science.gov (United States)

    2013-11-20

    ... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption; Extension of Comment Periods... Holding of Produce for Human Consumption'' and for its information collection provisions. DATES: The FDA... for Human Consumption.'' The original comment period of 120 days was extended several times and...

  12. 78 FR 17155 - Standards for the Growing, Harvesting, Packing, and Holding of Produce for Human Consumption...

    Science.gov (United States)

    2013-03-20

    ... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption; Correction AGENCY: Food and..., harvesting, packing, and holding of produce, meaning fruits and vegetables grown for human consumption. FDA..., Harvesting, Packing, and Holding of Produce for Human Consumption.'' The document published with several...

  13. 78 FR 48637 - Standards for the Growing, Harvesting, Packing, and Holding of Produce for Human Consumption...

    Science.gov (United States)

    2013-08-09

    ... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption; Extension of Comment Periods... Human Consumption,'' that appeared in the Federal Register of January 16, 2013. We are taking this... of Produce for Human Consumption'' with a 120-day comment period on the provisions of the proposed...

  14. Water resources in a rapidly growing region-Oakland County, Michigan

    Science.gov (United States)

    Aichele, Stephen S.

    2005-01-01

    Oakland County is a suburban county in southeast Michigan. Population and demand for water grew steadily in the county over the 20th century, and these trends are expected to continue in coming decades. Roughly 75 percent of current water demand is met by imported water from the Detroit Water and Sewerage Division (DWSD), but water use from ground-water sources within the county still exceeds 43 million gallons per day. Because much of the population growth is in areas beyond the DWSD system, an additional 20-25 million gallons per day of supply may be necessary to meet future demands. Managing the wastewater produced while also protecting human and ecosystem health also may present challenges.

  15. Peptostreptococcus productus strain that grows rapidly with CO as the energy source.

    Science.gov (United States)

    Lorowitz, W H; Bryant, M P

    1984-05-01

    Anaerobic bacteria were enriched with a sewage digestor sludge inoculum and a mineral medium supplemented with B-vitamins and 0.05% yeast extract and with a 50% CO-30% N2-20% CO2 (2 atm [202 kPa]) gas phase. Microscopic observation revealed an abundance of gram-positive cocci, 1.0 by 1.4 micron, which occurred in pairs or chains. The coccus, strain U-1, was isolated by using roll tubes with CO as the energy source. Based on morphology, sugars fermented, fermentation products from glucose (H2, acetate, lactate, and succinate), and other features, strain U-1 was identified as Peptostreptococcus productus IIb (similar to the type strain). The doubling time with up to 50% CO was 1.5 h; acetate and CO2 were the major products. In addition, no significant change in the doubling time was observed with 90% CO. Some stock strains were also able to use CO, although not as well. Strain U-1 produced acetate during growth with H2-CO2. Other C1 compounds did not support growth. Most probable numbers of CO utilizers morphologically identical with strain U-1 were 7.5 X 10(6) and 1.1 X 10(5) cells per g for anaerobic digestor sludge and human feces, respectively.

  16. The reasons for the rapid development of human technologies

    OpenAIRE

    Macfarlane, Alan

    2004-01-01

    While playing with a two-year-old girl in Australia, Alan Macfarlane explains one reason for the rapid development of humans, namely their curiosity and desire to experiment and to imagine and make new things.

  17. 78 FR 11611 - Standards for the Growing, Harvesting, Packing, and Holding of Produce for Human Consumption...

    Science.gov (United States)

    2013-02-19

    ... Growing, Harvesting, Packing, and Holding of Produce for Human Consumption; Extension of Comment Period... Consumption'' that appeared in the Federal Register of January 16, 2013. In the preamble to the proposed rule... of Produce for Human Consumption.'' FOR FURTHER INFORMATION CONTACT: Domini Bean, Office of...

  18. Rapidly growing Mycobacterium infections after cosmetic surgery in medical tourists: the Bronx experience and a review of the literature.

    Science.gov (United States)

    Cusumano, Lucas R; Tran, Vivy; Tlamsa, Aileen; Chung, Philip; Grossberg, Robert; Weston, Gregory; Sarwar, Uzma N

    2017-10-01

    Medical tourism is increasingly popular for elective cosmetic surgical procedures. However, medical tourism has been accompanied by reports of post-surgical infections due to rapidly growing mycobacteria (RGM). The authors' experience working with patients with RGM infections who have returned to the USA after traveling abroad for cosmetic surgical procedures is described here. Patients who developed RGM infections after undergoing cosmetic surgeries abroad and who presented at the Montefiore Medical Center (Bronx, New York, USA) between August 2015 and June 2016 were identified. A review of patient medical records was performed. Four patients who presented with culture-proven RGM infections at the sites of recent cosmetic procedures were identified. All patients were treated with a combination of antibiotics and aggressive surgical treatment. This case series of RGM infections following recent cosmetic surgeries abroad highlights the risks of medical tourism. Close monitoring of affected patients by surgical and infectious disease specialties is necessary, as aggressive surgical debridement combined with appropriate antibiotic regimens is needed to achieve cure. Given the increasing reports of post-surgical RGM infections, consultants should have a low threshold for suspecting RGM, as rapid diagnosis may accelerate the initiation of targeted treatment and minimize morbidity. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  19. Rapid evolution of the human mutation spectrum.

    Science.gov (United States)

    Harris, Kelley; Pritchard, Jonathan K

    2017-04-25

    DNA is a remarkably precise medium for copying and storing biological information. This high fidelity results from the action of hundreds of genes involved in replication, proofreading, and damage repair. Evolutionary theory suggests that in such a system, selection has limited ability to remove genetic variants that change mutation rates by small amounts or in specific sequence contexts. Consistent with this, using SNV variation as a proxy for mutational input, we report here that mutational spectra differ substantially among species, human continental groups and even some closely related populations. Close examination of one signal, an increased TCC→TTC mutation rate in Europeans, indicates a burst of mutations from about 15,000 to 2000 years ago, perhaps due to the appearance, drift, and ultimate elimination of a genetic modifier of mutation rate. Our results suggest that mutation rates can evolve markedly over short evolutionary timescales and suggest the possibility of mapping mutational modifiers.

  20. Methodology & Themes of Human-Robot Interaction: A Growing Research Field

    Directory of Open Access Journals (Sweden)

    Kerstin Dautenhahn

    2007-03-01

    Full Text Available This article discusses challenges of Human-Robot Interaction, which is a highly inter- and multidisciplinary area. Themes that are important in current research in this lively and growing field are identified and selected work relevant to these themes is discussed.

  1. Methodology & Themes of Human-Robot Interaction: A Growing Research Field

    OpenAIRE

    Dautenhahn, Kerstin

    2007-01-01

    This article discusses challenges of Human-Robot Interaction, which is a highly inter- and multidisciplinary area. Themes that are important in current research in this lively and growing field are identified and selected work relevant to these themes is discussed.

  2. Methodology & Themes of Human-Robot Interaction: A Growing Research Field

    OpenAIRE

    Kerstin Dautenhahn

    2008-01-01

    This article discusses challenges of Human-Robot Interaction, which is a highly inter- and multidisciplinary area. Themes that are important in current research in this lively and growing field are identified and selected work relevant to these themes is discussed.

  3. Methodology & Themes of Human-Robot Interaction: A Growing Research Field

    Directory of Open Access Journals (Sweden)

    Kerstin Dautenhahn

    2008-11-01

    Full Text Available This article discusses challenges of Human-Robot Interaction, which is a highly inter- and multidisciplinary area. Themes that are important in current research in this lively and growing field are identified and selected work relevant to these themes is discussed.

  4. Miniaturized extinction culturing is the preferred strategy for rapid isolation of fast‐growing methane‐oxidizing bacteria

    Science.gov (United States)

    Hoefman, Sven; van der Ha, David; De Vos, Paul; Boon, Nico; Heylen, Kim

    2012-01-01

    Summary Methane‐oxidizing bacteria (MOB) have a large potential as a microbial sink for the greenhouse gas methane as well as for biotechnological purposes. However, their application in biotechnology has so far been hampered, in part due to the relative slow growth rate of the available strains. To enable the availability of novel strains, this study compares the isolation of MOB by conventional dilution plating with miniaturized extinction culturing, both performed after an initial enrichment step. The extinction approach rendered 22 MOB isolates from four environmental samples, while no MOB could be isolated by plating. In most cases, extinction culturing immediately yielded MOB monocultures making laborious purification redundant. Both type I (Methylomonas spp.) and type II (Methylosinus sp.) MOB were isolated. The isolated methanotrophic diversity represented at least 11 different strains and several novel species based on 16S rRNA gene sequence dissimilarity. These strains possessed the particulate (100%) and soluble (64%) methane monooxygenase gene. Also, 73% of the strains could be linked to a highly active fast‐growing mixed MOB community. In conclusion, miniaturized extinction culturing was more efficient in rapidly isolating numerous MOB requiring little effort and fewer materials, compared with the more widely applied plating procedure. This miniaturized approach allowed straightforward isolation and could be very useful for subsequent screening of desired characteristics, in view of their future biotechnological potential. PMID:22070783

  5. Mycobacterium saopaulense sp. nov., a rapidly growing mycobacterium closely related to members of the Mycobacterium chelonae--Mycobacterium abscessus group.

    Science.gov (United States)

    Nogueira, Christiane Lourenço; Whipps, Christopher M; Matsumoto, Cristianne Kayoko; Chimara, Erica; Droz, Sara; Tortoli, Enrico; de Freitas, Denise; Cnockaert, Margo; Palomino, Juan Carlos; Martin, Anandi; Vandamme, Peter; Leão, Sylvia Cardoso

    2015-12-01

    Five isolates of non-pigmented, rapidly growing mycobacteria were isolated from three patients and,in an earlier study, from zebrafish. Phenotypic and molecular tests confirmed that these isolates belong to the Mycobacterium chelonae-Mycobacterium abscessus group, but they could not be confidently assigned to any known species of this group. Phenotypic analysis and biochemical tests were not helpful for distinguishing these isolates from other members of the M. chelonae–M.abscessus group. The isolates presented higher drug resistance in comparison with other members of the group, showing susceptibility only to clarithromycin. The five isolates showed a unique PCR restriction analysis pattern of the hsp65 gene, 100 % similarity in 16S rRNA gene and hsp65 sequences and 1-2 nt differences in rpoB and internal transcribed spacer (ITS) sequences.Phylogenetic analysis of a concatenated dataset including 16S rRNA gene, hsp65, and rpoB sequences from type strains of more closely related species placed the five isolates together, as a distinct lineage from previously described species, suggesting a sister relationship to a group consisting of M. chelonae, Mycobacterium salmoniphilum, Mycobacterium franklinii and Mycobacterium immunogenum. DNA–DNA hybridization values .70 % confirmed that the five isolates belong to the same species, while values ,70 % between one of the isolates and the type strains of M. chelonae and M. abscessus confirmed that the isolates belong to a distinct species. The polyphasic characterization of these isolates, supported by DNA–DNA hybridization results,demonstrated that they share characteristics with M. chelonae–M. abscessus members, butconstitute a different species, for which the name Mycobacterium saopaulense sp. nov. is proposed. The type strain is EPM10906T (5CCUG 66554T5LMG 28586T5INCQS 0733T).

  6. Mycobacterium stephanolepidis sp. nov., a rapidly growing species related to Mycobacterium chelonae, isolated from marine teleost fish, Stephanolepis cirrhifer.

    Science.gov (United States)

    Fukano, Hanako; Wada, Shinpei; Kurata, Osamu; Katayama, Kinya; Fujiwara, Nagatoshi; Hoshino, Yoshihiko

    2017-08-01

    A previously undescribed rapidly growing, non-pigmented mycobacterium was identified based on biochemical and nucleic acid analyses, as well as growth characteristics. Seven isolates were cultured from samples collected from five thread-sail filefish (Stephanolepis cirrhifer) and two farmed black scraper (Thamnaconus modestus). Bacterial growth occurred at 15-35 °C on Middlebrook 7H11 agar. The bacteria were positive for catalase activity at 68 °C and urease activity, intermediate for iron uptake, and negative for Tween 80 hydrolysis, nitrate reduction, semi-quantitative catalase activity and arylsulfatase activity at day 3. No growth was observed on Middlebrook 7H11 agar supplemented with picric acid, and very little growth was observed in the presence of 5 % NaCl. α- and α'-mycolates were identified in the cell walls, and a unique profile of the fatty acid methyl esters and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiles of the protein and cell-wall lipids were acquired. Sequence analysis revealed that the seven isolates shared identical sequences for the 16S rRNA, rpoB, hsp65, recA and sodA genes. Phylogenetic analysis of the five gene sequences confirmed that the isolates were unique, but closely related to Mycobacterium chelonae. Antibiotic susceptibility testing revealed the minimum inhibitory concentration (MIC) of clarithromycin against this novel species was Mycobacterium salmoniphilum. The hsp65 PCR restriction enzyme analysis pattern differed from those of M. chelonae and M. salmoniphilum. Based on these findings, the name Mycobacterium stephanolepidis sp. nov. is proposed for this novel species, with the type strain being NJB0901T (=JCM 31611T=KCTC 39843T).

  7. Mycobacterium saopaulense sp. nov., a rapidly growing mycobacterium closely related to members of the Mycobacterium chelonae–Mycobacterium abscessus group

    Science.gov (United States)

    Nogueira, Christiane Lourenço; Whipps, Christopher M.; Matsumoto, Cristianne Kayoko; Chimara, Erica; Droz, Sara; Tortoli, Enrico; de Freitas, Denise; Cnockaert, Margo; Palomino, Juan Carlos; Martin, Anandi; Vandamme, Peter

    2015-01-01

    Five isolates of non-pigmented, rapidly growing mycobacteria were isolated from three patients and, in an earlier study, from zebrafish. Phenotypic and molecular tests confirmed that these isolates belong to the Mycobacterium chelonae–Mycobacterium abscessus group, but they could not be confidently assigned to any known species of this group. Phenotypic analysis and biochemical tests were not helpful for distinguishing these isolates from other members of the M. chelonae–M. abscessus group. The isolates presented higher drug resistance in comparison with other members of the group, showing susceptibility only to clarithromycin. The five isolates showed a unique PCR restriction analysis pattern of the hsp65 gene, 100 % similarity in 16S rRNA gene and hsp65 sequences and 1–2 nt differences in rpoB and internal transcribed spacer (ITS) sequences. Phylogenetic analysis of a concatenated dataset including 16S rRNA gene, hsp65, and rpoB sequences from type strains of more closely related species placed the five isolates together, as a distinct lineage from previously described species, suggesting a sister relationship to a group consisting of M. chelonae, Mycobacterium salmoniphilum, Mycobacterium franklinii and Mycobacterium immunogenum. DNA–DNA hybridization values >70 % confirmed that the five isolates belong to the same species, while values isolates and the type strains of M. chelonae and M. abscessus confirmed that the isolates belong to a distinct species. The polyphasic characterization of these isolates, supported by DNA–DNA hybridization results, demonstrated that they share characteristics with M. chelonae–M. abscessus members, but constitute a different species, for which the name Mycobacterium saopaulense sp. nov. is proposed. The type strain is EPM 10906T ( = CCUG 66554T = LMG 28586T = INCQS 0733T). PMID:26358475

  8. Microsatellites evolve more rapidly in humans than in chimpanzees

    Energy Technology Data Exchange (ETDEWEB)

    Rubinsztein, D.C.; Leggo, J.; Amos, W. [Cambridge Univ. (United Kingdom)

    1995-12-10

    Microsatellites are highly polymorphic markers consisting of varying numbers of tandem repeats. At different loci, these repeats can consist of one to five nucleotides. Microsatellites have been used in many fields of genetics, including genetic mapping, linkage disequilibrium analyses, forensic studies, and population genetics. It is important that we understand their mutational processes better so that they can be exploited optimally for studies of human diversity and evolutionary genetics. We have analyzed 24 microsatellite loci in chimpanzees, East Anglians, and Sub-Saharan Africans. The stepwise-weighted genetic distances between the humans and the chimpanzees and between the two human populations were calculated according to the method described by Deka et al. The ratio of the genetic distances between the chimpanzees and the humans relative to that between the Africans and the East Anglians was more than 10 times smaller than expected. This suggests that microsatellites have evolved more rapidly in humans than in chimpanzees. 12 refs., 1 tab.

  9. Rapid neurogenesis through transcriptional activation in human stem cells.

    Science.gov (United States)

    Busskamp, Volker; Lewis, Nathan E; Guye, Patrick; Ng, Alex H M; Shipman, Seth L; Byrne, Susan M; Sanjana, Neville E; Murn, Jernej; Li, Yinqing; Li, Shangzhong; Stadler, Michael; Weiss, Ron; Church, George M

    2014-11-17

    Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human-induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems-level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types. © 2014 The Authors. Published under the terms of the CC BY 4.0 license.

  10. Rapid Prototyping Human Interfaces Using Stretchable Strain Sensor

    Directory of Open Access Journals (Sweden)

    Tokiya Yamaji

    2017-01-01

    Full Text Available In the modern society with a variety of information electronic devices, human interfaces increase their importance in a boundary of a human and a device. In general, the human is required to get used to the device. Even if the device is designed as a universal device or a high-usability device, the device is not suitable for all users. The usability of the device depends on the individual user. Therefore, personalized and customized human interfaces are effective for the user. To create customized interfaces, we propose rapid prototyping human interfaces using stretchable strain sensors. The human interfaces comprise parts formed by a three-dimensional printer and the four strain sensors. The three-dimensional printer easily makes customized human interfaces. The outputs of the interface are calculated based on the sensor’s lengths. Experiments evaluate three human interfaces: a sheet-shaped interface, a sliding lever interface, and a tilting lever interface. We confirm that the three human interfaces obtain input operations with a high accuracy.

  11. Genetics of rapid eye movement sleep in humans

    OpenAIRE

    Adamczyk, M.; Ambrosius, U; Lietzenmaier, S; Wichniak, A; Holsboer, F.; Friess, E

    2015-01-01

    The trait-like nature of electroencephalogram (EEG) is well established. Furthermore, EEG of wake and non-rapid eye movement (non-REM) sleep has been shown to be highly heritable. However, the genetic effects on REM sleep EEG microstructure are as yet unknown. REM sleep is of special interest since animal and human data suggest a connection between REM sleep abnormalities and the pathophysiology of psychiatric and neurological diseases. Here we report the results of a study in monozygotic (MZ...

  12. Rapid Functional Reorganization in Human Cortex Following Neural Perturbation

    OpenAIRE

    Zanto, Theodore P.; Chadick, James Z.; Satris, Gabriela; Gazzaley, Adam

    2013-01-01

    Despite the human brain's ability to rapidly reorganize neuronal activity patterns in response to interactions with the environment (e.g., learning), it remains unclear whether compensatory mechanisms occur, on a similar time scale, in response to exogenous cortical perturbations. To investigate this, we disrupted normal neural function via repetitive transcranial magnetic stimulation and assessed, using fMRI, activity changes associated with performance on a working memory task. Although tra...

  13. Rapid induction of senescence in human cervical carcinoma cells

    Science.gov (United States)

    Goodwin, Edward C.; Yang, Eva; Lee, Chan-Jae; Lee, Han-Woong; Dimaio, Daniel; Hwang, Eun-Seong

    2000-09-01

    Expression of the bovine papillomavirus E2 regulatory protein in human cervical carcinoma cell lines repressed expression of the resident human papillomavirus E6 and E7 oncogenes and within a few days caused essentially all of the cells to synchronously display numerous phenotypic markers characteristic of cells undergoing replicative senescence. This process was accompanied by marked but in some cases transient alterations in the expression of cell cycle regulatory proteins and by decreased telomerase activity. We propose that the human papillomavirus E6 and E7 proteins actively prevent senescence from occurring in cervical carcinoma cells, and that once viral oncogene expression is extinguished, the senescence program is rapidly executed. Activation of endogenous senescence pathways in cancer cells may represent an alternative approach to treat human cancers.

  14. Machine takeover the growing threat to human freedom in a computer-controlled society

    CERN Document Server

    George, Frank Honywill

    1977-01-01

    Machine Takeover: The Growing Threat to Human Freedom in a Computer-Controlled Society discusses the implications of technological advancement. The title identifies the changes in society that no one is aware of, along with what this changes entails. The text first covers the information science, particularly the aspect of an automated system for information processing. Next, the selection deals with social implications of information science, such as information pollution. The text also tackles the concerns in the utilization of technology in order to manipulate the lives of people without th

  15. Diet rapidly and reproducibly alters the human gut microbiome

    Science.gov (United States)

    David, Lawrence A.; Maurice, Corinne F.; Carmody, Rachel N.; Gootenberg, David B.; Button, Julie E.; Wolfe, Benjamin E.; Ling, Alisha V.; Devlin, A. Sloan; Varma, Yug; Fischbach, Michael A.; Biddinger, Sudha B.; Dutton, Rachel J.; Turnbaugh, Peter J.

    2013-01-01

    Long-term diet influences the structure and activity of the trillions of microorganisms residing in the human gut1–5, but it remains unclear how rapidly and reproducibly the human gut microbiome responds to short-term macronutrient change. Here, we show that the short-term consumption of diets composed entirely of animal or plant products alters microbial community structure and overwhelms inter-individual differences in microbial gene expression. The animal-based diet increased the abundance of bile-tolerant microorganisms (Alistipes, Bilophila, and Bacteroides) and decreased the levels of Firmicutes that metabolize dietary plant polysaccharides (Roseburia, Eubacterium rectale, and Ruminococcus bromii). Microbial activity mirrored differences between herbivorous and carnivorous mammals2, reflecting trade-offs between carbohydrate and protein fermentation. Foodborne microbes from both diets transiently colonized the gut, including bacteria, fungi, and even viruses. Finally, increases in the abundance and activity of Bilophila wadsworthia on the animal-based diet support a link between dietary fat, bile acids, and the outgrowth of microorganisms capable of triggering inflammatory bowel disease6. In concert, these results demonstrate that the gut microbiome can rapidly respond to altered diet, potentially facilitating the diversity of human dietary lifestyles. PMID:24336217

  16. Rapid molecular identification of human taeniid cestodes by pyrosequencing approach.

    Directory of Open Access Journals (Sweden)

    Tongjit Thanchomnang

    Full Text Available Taenia saginata, T. solium, and T. asiatica are causative agents of taeniasis in humans. The difficulty of morphological identification of human taeniids can lead to misdiagnosis or confusion. To overcome this problem, several molecular methods have been developed, but use of these tends to be time-consuming. Here, a rapid and high-throughput pyrosequencing approach was developed for the identification of three human taeniids originating from various countries. Primers targeting the mitochondrial cytochrome c oxidase subunit 1 (cox1 gene of the three Taenia species were designed. Variations in a 26-nucleotide target region were used for identification. The reproducibility and accuracy of the pyrosequencing technology was confirmed by Sanger sequencing. This technique will be a valuable tool to distinguish between sympatric human taeniids that occur in Thailand, Asia and Pacific countries. This method could potentially be used for the molecular identification of the taeniid species that might be associated with suspicious cysts and lesions, or cyst residues in humans or livestock at the slaughterhouse.

  17. Rapid Molecular Identification of Human Taeniid Cestodes by Pyrosequencing Approach

    Science.gov (United States)

    Thanchomnang, Tongjit; Tantrawatpan, Chairat; Intapan, Pewpan M.; Sanpool, Oranuch; Janwan, Penchom; Lulitanond, Viraphong; Tourtip, Somjintana; Yamasaki, Hiroshi; Maleewong, Wanchai

    2014-01-01

    Taenia saginata, T. solium, and T. asiatica are causative agents of taeniasis in humans. The difficulty of morphological identification of human taeniids can lead to misdiagnosis or confusion. To overcome this problem, several molecular methods have been developed, but use of these tends to be time-consuming. Here, a rapid and high-throughput pyrosequencing approach was developed for the identification of three human taeniids originating from various countries. Primers targeting the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of the three Taenia species were designed. Variations in a 26-nucleotide target region were used for identification. The reproducibility and accuracy of the pyrosequencing technology was confirmed by Sanger sequencing. This technique will be a valuable tool to distinguish between sympatric human taeniids that occur in Thailand, Asia and Pacific countries. This method could potentially be used for the molecular identification of the taeniid species that might be associated with suspicious cysts and lesions, or cyst residues in humans or livestock at the slaughterhouse. PMID:24945530

  18. Rapid and Decentralized Human Waste Treatment by Microwave Radiation.

    Science.gov (United States)

    Nguyen, Tu Anh; Babel, Sandhya; Boonyarattanakalin, Siwarutt; Koottatep, Thammarat

    2017-07-01

      This study evaluates the technical feasibility of using microwave radiation for the rapid treatment of human feces. Human feces of 1000 g were radiated with a commercially available household microwave oven (with rotation) at different exposure time lengths (30, 50, 60, 70, and 75 mins) and powers (600, 800, and 1000 W). Volume reduction over 90% occurred after 1000 W microwave radiation for 75 mins. Pathogen eradiation performances of six log units or more at a high range of microwave powers were achieved. Treatments with the same energy input of 1000 Wh, but at lower powers with prolonged exposure times, significantly enhanced moisture removal and volume reduction. Microwave radiation caused carbonization and resulted in a more stable end product. The energy content of the samples after microwave treatment at 1000 W and 75 mins is 3517 ± 8.85 calories/g of dried sample, and the product can also be used as compost.

  19. How rapidly does the human mitochondrial genome evolve?

    Energy Technology Data Exchange (ETDEWEB)

    Howell, N.; Kubacka, I. [Univ. of Texas Medical Branch, Galveston, TX (United States); Mackey, D.A. [Univ. of Melbourne (Australia)]|[Univ. of Tasmania, Launceston (Australia)

    1996-09-01

    The results of an empirical nucleotide-sequencing approach indicate that the evolution of the human mitochondrial noncoding D-loop is both more rapid and more complex than is revealed by standard phylogenetic approaches. The nucleotide sequence of the D-loop region of the mitochondrial genome was determined for 45 members of a large matrilineal Leber hereditary optic neuropathy pedigree. Two germ-line mutations have arisen in members of one branch of the family, thereby leading to triplasmic descendants with three mitochondrial genotypes. Segregation toward the homoplasmic state can occur within a single generation in some of these descendants, a result that suggests rapid fixation of mitochondrial mutations as a result of developmental bottlenecking. However, slow segregation was observed in other offspring, and therefore no single or simple pattern of segregation can be generalized from the available data. Evidence for rare mtDNA recombination within the D-loop was obtained for one family member. In addition to these germ-line mutations, a somatic mutation was found in the D-loop of one family member. When this genealogical approach was applied to the nucleotide sequences of mitochondrial coding regions, the results again indicated a very rapid rate of evolution. 44 refs., 2 figs., 2 tabs.

  20. Rapid local anesthesia in humans using minimally invasive microneedles.

    Science.gov (United States)

    Gupta, Jyoti; Denson, Donald D; Felner, Eric I; Prausnitz, Mark R

    2012-02-01

    This study tested the hypothesis that minimally invasive microneedles cause less pain during injection of lidocaine, but induce local anesthesia in humans with the same rapid onset and efficacy as intradermal lidocaine injection using hypodermic needles. This study was a randomized, single-blinded, within participants, controlled design. Hollow, 500-μm long microneedles were used to inject lidocaine to the forearm of 15 human participants. The associated pain was recorded using a visual analog (VAS) scale. The area and depth of numbness were determined at 0, 7.5, and 15 minutes after injection. Lidocaine was also injected to the dorsum of the hand near a vein, followed by placement of an intravenous catheter and measurement of associated pain. A 26-gauge intradermal bevel hypodermic needle similarly administered lidocaine on the opposite forearm/hand to serve as the positive control. VAS pain scores revealed that injection using microneedles was significantly less painful than hypodermic needles for both the forearm and dorsum of the hand injections. However, there was no significant difference in the area or depth of the resulting numbness between the 2 treatment methods at any time point (0, 7.5, and 15 min) indicating that microneedles had immediate onset and were as effective as hypodermic needles in inducing dermal anesthesia. Moreover, insertion of an intravenous catheter immediately after lidocaine injection on the dorsum of the hand led to comparable pain scores for the microneedle and hypodermic needle treated sites, further confirming efficacy of microneedles in inducing rapid local anesthesia. Lastly, 77% of the participants preferred microneedles and 80% indicated that they did not consider microneedles to be painful. This study demonstrates for the first time that microneedle-based lidocaine injection is as rapid and as effective as hypodermic injection in inducing local anesthesia while resulting in significantly less pain during injection.

  1. Planetary health: protecting human health on a rapidly changing planet.

    Science.gov (United States)

    Myers, Samuel S

    2018-12-23

    The impact of human activities on our planet's natural systems has been intensifying rapidly in the past several decades, leading to disruption and transformation of most natural systems. These disruptions in the atmosphere, oceans, and across the terrestrial land surface are not only driving species to extinction, they pose serious threats to human health and wellbeing. Characterising and addressing these threats requires a paradigm shift. In a lecture delivered to the Academy of Medical Sciences on Nov 13, 2017, I describe the scale of human impacts on natural systems and the extensive associated health effects across nearly every dimension of human health. I highlight several overarching themes that emerge from planetary health and suggest advances in the way we train, reward, promote, and fund the generation of health scientists who will be tasked with breaking out of their disciplinary silos to address this urgent constellation of health threats. I propose that protecting the health of future generations requires taking better care of Earth's natural systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Regulation of the efflux of putrescine and cadaverine from rapidly growing cultured RAW 264 cells by extracellular putrescine.

    OpenAIRE

    Tjandrawinata, R R; Byus, C V

    1995-01-01

    Cultures of the macrophage-like RAW 264 cells were adapted to divide normally in a synthetic serum-supplemented culture medium lacking any polyamines and diamine oxidase activity. These rapidly dividing cells actively effluxed large amounts of putrescine and cadaverine, compared with the intracellular levels, into the culture medium. The efflux of putrescine was stimulated by the amino acid ornithine, whereas efflux of cadaverine was inhibited. Relatively low levels of spermidine and N1-acety...

  3. Rapidly growing Mycobacterium infections after cosmetic surgery in medical tourists: the Bronx experience and a review of the literature

    Directory of Open Access Journals (Sweden)

    Lucas R. Cusumano

    2017-10-01

    Conclusions: This case series of RGM infections following recent cosmetic surgeries abroad highlights the risks of medical tourism. Close monitoring of affected patients by surgical and infectious disease specialties is necessary, as aggressive surgical debridement combined with appropriate antibiotic regimens is needed to achieve cure. Given the increasing reports of post-surgical RGM infections, consultants should have a low threshold for suspecting RGM, as rapid diagnosis may accelerate the initiation of targeted treatment and minimize morbidity.

  4. Do farmers rapidly adapt to past growing conditions by sowing different proportions of early and late maturing cereals and cultivars?

    Directory of Open Access Journals (Sweden)

    Pirjo Peltonen-Sainio

    2013-10-01

    Full Text Available In the short growing season of the northernmost European growing conditions, farmers are increasingly interested in expanding cultivation of later maturing crops at the expense of early maturing ones with lower yields. In this study we aimed to assess how the switching between spring cereals that differ in earliness was associated with different external factors. This was tested using unique datasets for regional cropping areas and cultivar use for the last 15 years. Early maturing barley was favored at the expense of later maturing wheat when a high number of days to crop maturity was required in the preceding year. In contrast, farmers reduced the barley area when a high number of cumulated degree days was required for a crop to mature in the previous year. A shift was recorded from early to late maturing cultivars. This study indicated that despite limited opportunities for farmers to alter land use, they readily responded to past conditions and used the knowledge gained for decision-making to reduce risk. This is a valuable operative model for studying adaptation to opportunities and constraints induced by climate change.

  5. Monitoring Annual Urban Changes in a Rapidly Growing Portion of Northwest Arkansas with a 20-Year Landsat Record

    Directory of Open Access Journals (Sweden)

    Ryan Reynolds

    2017-01-01

    Full Text Available Northwest Arkansas has undergone a significant urban transformation in the past several decades and is considered to be one of the fastest growing regions in the United States. The urban area expansion and the associated demographic increases bring unprecedented pressure to the environment and natural resources. To better understand the consequences of urbanization, accurate and long-term depiction on urban dynamics is critical. Although urban mapping activities using remote sensing have been widely conducted, long-term urban growth mapping at an annual pace is rare and the low accuracy of change detection remains a challenge. In this study, a time series Landsat stack covering the period from 1995 to 2015 was employed to detect the urban dynamics in Northwest Arkansas via a two-stage classification approach. A set of spectral indices that have been proven to be useful in urban area extraction together with the original Landsat spectral bands were used in the maximum likelihood classifier and random forest classifier to distinguish urban from non-urban pixels for each year. A temporal trajectory polishing method, involving temporal filtering and heuristic reasoning, was then applied to the sequence of classified urban maps for further improvement. Based on a set of validation samples selected for five distinct years, the average overall accuracy of the final polished maps was 91%, which improved the preliminary classifications by over 10%. Moreover, results from this study also indicated that the temporal trajectory polishing method was most effective with initial low accuracy classifications. The resulting urban dynamic map is expected to provide unprecedented details about the area, spatial configuration, and growing trends of urban land-cover in Northwest Arkansas.

  6. Using housing growth to estimate habitat change: detecting Ovenbird response in a rapidly growing New England State

    Science.gov (United States)

    Christopher A. Lepczyk; Aaron Wunnicke; Volker C. Radeloff; Curtis H. Flather; Anna M. Pidgeon; Roger B. Hammer

    2013-01-01

    Numerous measures of human influence on the environment exist, but one that is of particular importance is houses as they can impact the environment from species through the landscape level. Furthermore, because the addition of houses represents an important component of landscape change, housing information could be used to assess ecological responses (e.g., decline...

  7. Growing City and Rapid Land Use Transition: Assessing Multiple Hazards and Risks in the Pokhara Valley, Nepal

    Directory of Open Access Journals (Sweden)

    Bhagawat Rimal

    2015-10-01

    Full Text Available Pokhara is one of the most naturally beautiful cities in the world with a unique geological setting. This important tourist city is under intense pressure from rapid urbanization and population growth. Multiple hazards and risks are rapidly increasing in Pokhara due to unsustainable land use practices, particularly the increase in built-up areas. This study examines the relationship among urbanization, land use/land cover dynamics and multiple hazard and risk analysis of the Pokhara valley from 1990 to 2013. We investigate some of the active hazards, such as floods, landslides, fire, sinkholes, land subsidence and earthquakes, and prepare an integrated multiple hazard risk map indicating the highly vulnerable zones. Land use and land cover maps from 1990 and 2013 from Landsat images (30 m resolution have been prepared and analyzed for the spatial dynamics of urbanization and the transition of land use and land cover. In the 23-year period, the built-up area more than doubled from 24.03 km² to 54.20 km². Although the landscape in the urban, peri-urban and rural areas appears to be fragmented, different drivers play pivotal roles in landscape change in these areas. The results provide substantial information for establishing innovative action plans for disaster risk management in the valley. Recommendations are made for the most suitable places for future urban expansion in the valley. This study is important for raising awareness among policy makers and other public officials to include multiple hazard risk mitigation in land use policies and plans. Establishing connections between urban expansions, escalating population growth and multiple hazards and risk assessment will also improve in modelling the latent impact of future catastrophes and emergency preparedness.

  8. Mycobacterium talmoniae sp. nov., a slowly growing mycobacterium isolated from human respiratory samples.

    Science.gov (United States)

    Davidson, Rebecca M; DeGroote, Mary Ann; Marola, Jamie L; Buss, Sarah; Jones, Victoria; McNeil, Michael R; Freifeld, Alison G; Elaine Epperson, L; Hasan, Nabeeh A; Jackson, Mary; Iwen, Peter C; Salfinger, Max; Strong, Michael

    2017-08-01

    A novel slowly growing, non-chromogenic species of the class Actinobacteria was isolated from a human respiratory sample in Nebraska, USA, in 2012. Analysis of the internal transcribed spacer sequence supported placement into the genus Mycobacterium with high sequence similarity to a previously undescribed strain isolated from a patient respiratory sample from Oregon, USA, held in a collection in Colorado, USA, in 2000. The two isolates were subjected to phenotypic testing and whole genome sequencing and found to be indistinguishable. The bacteria were acid-fast stain-positive, rod-shaped and exhibited growth after 7-10 days on solid media at temperatures ranging from 25 to 42°C. Colonies were non-pigmented, rough and slightly raised. Analyses of matrix-assisted laser desorption ionization time-of-flight profiles showed no matches against a reference library of 130 mycobacterial species. Full-length 16S rRNA gene sequences were identical for the two isolates, the average nucleotide identity (ANI) between their genomes was 99.7 % and phylogenetic comparisons classified the novel mycobacteria as the basal most species in the slowly growing Mycobacterium clade. Mycobacterium avium is the most closely related species based on rpoB gene sequence similarity (92 %), but the ANI between the genomes was 81.5 %, below the suggested cut-off for differentiating two species (95 %). Mycolic acid profiles were more similar to M. avium than to Mycobacterium simiae or Mycobacterium abscessus. The phenotypic and genomic data support the conclusion that the two related isolates represent a novel Mycobacterium species for which the name Mycobacterium talmoniae sp. nov. is proposed. The type strain is NE-TNMC-100812T (=ATCC BAA-2683T=DSM 46873T).

  9. Rapid extraction and preservation of genomic DNA from human samples.

    Science.gov (United States)

    Kalyanasundaram, D; Kim, J-H; Yeo, W-H; Oh, K; Lee, K-H; Kim, M-H; Ryew, S-M; Ahn, S-G; Gao, D; Cangelosi, G A; Chung, J-H

    2013-02-01

    Simple and rapid extraction of human genomic DNA remains a bottleneck for genome analysis and disease diagnosis. Current methods using microfilters require cumbersome, multiple handling steps in part because salt conditions must be controlled for attraction and elution of DNA in porous silica. We report a novel extraction method of human genomic DNA from buccal swab and saliva samples. DNA is attracted onto a gold-coated microchip by an electric field and capillary action while the captured DNA is eluted by thermal heating at 70 °C. A prototype device was designed to handle four microchips, and a compatible protocol was developed. The extracted DNA using microchips was characterized by qPCR for different sample volumes, using different lengths of PCR amplicon, and nuclear and mitochondrial genes. In comparison with a commercial kit, an equivalent yield of DNA extraction was achieved with fewer steps. Room-temperature preservation for 1 month was demonstrated for captured DNA, facilitating straightforward collection, delivery, and handling of genomic DNA in an environment-friendly protocol.

  10. An aerosol climatology for a rapidly growing arid region (southern Arizona): Major aerosol species and remotely sensed aerosol properties.

    Science.gov (United States)

    Sorooshian, Armin; Wonaschütz, Anna; Jarjour, Elias G; Hashimoto, Bryce I; Schichtel, Bret A; Betterton, Eric A

    2011-10-01

    This study reports a comprehensive characterization of atmospheric aerosol particle properties in relation to meteorological and back trajectory data in the southern Arizona region, which includes two of the fastest growing metropolitan areas in the United States (Phoenix and Tucson). Multiple data sets (MODIS, AERONET, OMI/TOMS, MISR, GOCART, ground-based aerosol measurements) are used to examine monthly trends in aerosol composition, aerosol optical depth (AOD), and aerosol size. Fine soil, sulfate, and organics dominate PM2.5 mass in the region. Dust strongly influences the region between March and July owing to the dry and hot meteorological conditions and back trajectory patterns. Because monsoon precipitation begins typically in July, dust levels decrease, while AOD, sulfate, and organic aerosol reach their maximum levels because of summertime photochemistry and monsoon moisture. Evidence points to biogenic volatile organic compounds being a significant source of secondary organic aerosol in this region. Biomass burning also is shown to be a major contributor to the carbonaceous aerosol budget in the region, leading to enhanced organic and elemental carbon levels aloft at a sky-island site north of Tucson (Mt. Lemmon). Phoenix exhibits different monthly trends for aerosol components in comparison with the other sites owing to the strong influence of fossil carbon and anthropogenic dust. Trend analyses between 1988 and 2009 indicate that the strongest statistically significant trends are reductions in sulfate, elemental carbon, and organic carbon, and increases in fine soil during the spring (March-May) at select sites. These results can be explained by population growth, land-use changes, and improved source controls.

  11. A rapid DNA extraction method suitable for human papillomavirus detection.

    Science.gov (United States)

    Brestovac, Brian; Wong, Michelle E; Costantino, Paul S; Groth, David

    2014-04-01

    Infection with oncogenic human papillomavirus (HPV) genotypes is necessary for the development of cervical cancer. Testing for HPV DNA from liquid based cervical samples can be used as an adjunct to traditional cytological screening. In addition there are ongoing viral load, genotyping, and prevalence studies. Therefore, a sensitive DNA extraction method is needed to maximize the efficiency of HPV DNA detection. The XytXtract Tissue kit is a DNA extraction kit that is rapid and so could be useful for HPV testing, particularly in screening protocols. This study was undertaken to determine the suitability of this method for HPV detection. DNA extraction from HeLa and Caski cell lines containing HPV 18 and 16 respectively together with DNA from five liquid based cervical samples were used in a HPV PCR assay. DNA was also extracted using the QIAamp DNA mini kit (Qiagen, Hilden, Germany) as a comparison. DNA extracts were serially diluted and assayed. HPV DNA was successfully detected in cell lines and cervical samples using the XytXtract Tissue kit. In addition, the XytXtract method was found to be more sensitive than the QIAmp method as determined by a dilution series of the extracted DNA. While the XytXtract method is a closed, the QIAamp method uses a spin column with possible loss of DNA through DNA binding competition of the matrix, which could impact on the final extraction efficiency. The XytXtract is a cheap, rapid and efficient method for extracting HPV DNA from both cell lines and liquid based cervical samples. © 2014 Wiley Periodicals, Inc.

  12. [Rapid diagnostic tests for the serodiagnosis of human cystic echinococcosis].

    Science.gov (United States)

    Tamarozzi, F; Mariconti, M; Covini, I; Brunetti, E

    2017-02-01

    Cystic echinococcosis (CE) is a parasitic zoonosis especially affecting resource-poor populations in livestock raising areas. Imaging, in particular ultrasound (US), is crucial for the diagnosis, staging, and clinical management of abdominal CE in humans. Serology is a valuable complement to imaging, especially when ultrasound features of CE are absent or unclear. In rural endemic areas, where expertise in US is scant, and conventional serology techniques are unavailable due to lack of laboratory equipment, rapid diagnostic tests (RDTs) may be very useful. Several reports have described the performance of commercial and experimental RDTs in the diagnosis of CE, including a recent study by our group that compared the diagnostic performances of three commercial RDTs for the diagnosis of hepatic CE. To put RDTs for CE in context, we reviewed the available literature in English on this topic. Overall, RDTs appear to be useful in resourcepoor settings where they may replace conventional serodiagnostic tests. However, like other serodiagnostic tests, RDTs lack standardization and show unsatisfactory sensitivity and specificity. An important issue that needs to be addressed is that studies on the diagnostic performance of RDTs fail to take into account the variables known to influence results such as anatomical location and cyst stage.

  13. Rapid Detection of Human Norovirus in Frozen Raspberries.

    Science.gov (United States)

    Summa, Maija; Maunula, Leena

    2017-10-10

    Raspberries have lately caused several human norovirus (HuNoV) outbreaks in Europe. In this study, we developed and evaluated for HuNoV reverse transcription (RT)-PCR detection in frozen raspberries extraction methods that have equal sensitivity but are less time-consuming than widely used methods based on polyethylene glycol (PEG) precipitation and chloroform-butanol purification. One method was applied to stored frozen raspberries linked to previous HuNoV outbreaks and berries on sale. In the virus elution-based Method 1, sparkling water eluted viruses most efficiently from the berries. Method 2, based on direct nucleic acid extraction with minor PEG supplement, yielded the highest number of positive findings (4 out of 9) at low virus concentration level of 100 genome copies HuNoV genogroup II per 25 g raspberries. Both methods showed approximately equal sensitivity to a method including PEG precipitation and chloroform-butanol purification. Two naturally contaminated berry samples linked to HuNoV outbreaks in 2006 and 2009 were still positive for HuNoV genogroup I, but all berry products purchased from a local store remained negative for HuNoV. In conclusion, this study presents two efficient and rapid methods which can be used in urgent HuNoV outbreak investigations, since the results of the virus analysis are available in a few hours.

  14. [Rapid growing liposarcoma in retroperitoneum

    DEFF Research Database (Denmark)

    Engel, L.E.; Mynster, T.

    2008-01-01

    inhomogeneous tumour adhesive to the right kidney capsule was removed in toto from retroperitoneum. Histological diagnosis: Sclerosing, low differentiated liposarcoma. Besides radical surgery only radiation therapy may have effect in treatment of retroperitoneal soft tissue sarcomas Udgivelsesdato: 2008/9/29...

  15. Human domination of the biosphere: Rapid discharge of the earth-space battery foretells the future of humankind.

    Science.gov (United States)

    Schramski, John R; Gattie, David K; Brown, James H

    2015-08-04

    Earth is a chemical battery where, over evolutionary time with a trickle-charge of photosynthesis using solar energy, billions of tons of living biomass were stored in forests and other ecosystems and in vast reserves of fossil fuels. In just the last few hundred years, humans extracted exploitable energy from these living and fossilized biomass fuels to build the modern industrial-technological-informational economy, to grow our population to more than 7 billion, and to transform the biogeochemical cycles and biodiversity of the earth. This rapid discharge of the earth's store of organic energy fuels the human domination of the biosphere, including conversion of natural habitats to agricultural fields and the resulting loss of native species, emission of carbon dioxide and the resulting climate and sea level change, and use of supplemental nuclear, hydro, wind, and solar energy sources. The laws of thermodynamics governing the trickle-charge and rapid discharge of the earth's battery are universal and absolute; the earth is only temporarily poised a quantifiable distance from the thermodynamic equilibrium of outer space. Although this distance from equilibrium is comprised of all energy types, most critical for humans is the store of living biomass. With the rapid depletion of this chemical energy, the earth is shifting back toward the inhospitable equilibrium of outer space with fundamental ramifications for the biosphere and humanity. Because there is no substitute or replacement energy for living biomass, the remaining distance from equilibrium that will be required to support human life is unknown.

  16. Humans copy rapidly increasing choices in a multiarmed bandit problem

    NARCIS (Netherlands)

    Toelch, U.; Bruce, M.J.; Meeus, M.T.H.; Reader, S.M.

    2010-01-01

    Conformist social learning, the tendency to acquire the most common trait in a group, allows individuals to rapidly acquire established beneficial traits from a multitude of options. However, conformist strategies hinder acquisition of novel advantageous behavior patterns, because such innovations

  17. Rapid changes in the gut microbiome during human evolution.

    Science.gov (United States)

    Moeller, Andrew H; Li, Yingying; Mpoudi Ngole, Eitel; Ahuka-Mundeke, Steve; Lonsdorf, Elizabeth V; Pusey, Anne E; Peeters, Martine; Hahn, Beatrice H; Ochman, Howard

    2014-11-18

    Humans are ecosystems containing trillions of microorganisms, but the evolutionary history of this microbiome is obscured by a lack of knowledge about microbiomes of African apes. We sequenced the gut communities of hundreds of chimpanzees, bonobos, and gorillas and developed a phylogenetic approach to reconstruct how present-day human microbiomes have diverged from those of ancestral populations. Compositional change in the microbiome was slow and clock-like during African ape diversification, but human microbiomes have deviated from the ancestral state at an accelerated rate. Relative to the microbiomes of wild apes, human microbiomes have lost ancestral microbial diversity while becoming specialized for animal-based diets. Individual wild apes cultivate more phyla, classes, orders, families, genera, and species of bacteria than do individual humans across a range of societies. These results indicate that humanity has experienced a depletion of the gut flora since diverging from Pan.

  18. Do teachers and students get the Ed-Tech products they need: The challenges of Ed-Tech procurement in a rapidly growing market

    Directory of Open Access Journals (Sweden)

    Jennifer Morrison

    2015-03-01

    Full Text Available Ed-tech courseware products to support teaching and learning are being developed and made available for acquisition by school districts at a rapid rate. In this growing market, developers and providers face challenges with making their products visible to customers, while school district stakeholders must grapple with “discovering” which products of the many available best address their instructional needs. The present study presents the experiences with and perceptions about the procurement process from 47 superintendents representing diverse school districts in the U. S. Results indicate that, while improvements are desired in many aspects of the procurement process, the superintendents, overall, believe that, once desired products are identified, they are generally able to acquire them. Difficulties lie in tighter budgets, discovering products that are potentially the best choices, and evaluating the effectiveness of the products selected as options. These findings are presented and interpreted in relation to five major “Action Points” in the procurement process, and also with regard to implications for evaluating how educational technology impacts K-12 instruction.

  19. In Vitro Comparison of Ertapenem, Meropenem, and Imipenem against Isolates of Rapidly Growing Mycobacteria and Nocardia by Use of Broth Microdilution and Etest.

    Science.gov (United States)

    Brown-Elliott, Barbara A; Killingley, Jessica; Vasireddy, Sruthi; Bridge, Linda; Wallace, Richard J

    2016-06-01

    We compared the activities of the carbapenems ertapenem, meropenem, and imipenem against 180 isolates of rapidly growing mycobacteria (RGM) and 170 isolates of Nocardia using the Clinical and Laboratory Standards Institute (CLSI) guidelines. A subset of isolates was tested using the Etest. The rate of susceptibility to ertapenem and meropenem was limited and less than that to imipenem for the RGM. Analysis of major and minor discrepancies revealed that >90% of the isolates of Nocardia had higher MICs by the broth microdilution method than by Etest, in contrast to the lower broth microdilution MICs seen for >80% of the RGM. Imipenem remains the most active carbapenem against RGM, including Mycobacterium abscessus subsp. abscessus For Nocardia, imipenem was significantly more active only against Nocardia farcinica Although there may be utility in testing the activities of the newer carbapenems against Nocardia, their activities against the RGM should not be routinely tested. Testing by Etest is not recommended by the CLSI. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  20. Comparison of the immature sheep spine and the growing human spine: a spondylometric database for growth modulating research.

    Science.gov (United States)

    Hasler, Carol; Sprecher, Christoph Martin; Milz, Stefan

    2010-11-01

    A comparative study on growth of the sheep and human spine. To validate the immature sheep spine as model for the growing human spine and to yield a database for planning and interpretation of future animal experiments. With the current change of paradigm to nonfusion strategies for pediatric spine deformities, experimental surgery on spines of growing goats, sheep, and pigs has gained importance as preclinical proof-of-concept test. However, despite the proceeding use of animals, there is a lack of knowledge regarding the growth of the sheep spine and the relation to the human spine. Thoracic and lumbar cadaver spines were harvested from 50 Swiss alpine sheep. Specimens were obtained from newborn, 1, 3, 6, 9 and 12, 15 and 18 months old female sheep. Direct spondylometry yielded vertebral body heights, widths, and depths and spinal canal size, which were compared to pooled data on human spine growth retrieved from the literature. Sheep spine growth ceases at age 15 to 18 months, which corresponds to a time-lapse model of human growth. Main growth occurs within the first 3 to 6 months of life, as opposed to human spines with maximal growth during the first 4 years and puberty. The relation between sheep and human vertebral shape is continuously changing with growth: at birth, sheep vertebrae are twice as tall, but equally wide and deep. At skeletal maturity, height is 15% to 25% bigger in sheep, but width 15% to 30% and depth 30% to 50% are smaller. The immature sheep spine offers fast effects if growth-modulating interventions are performed within the first 3 to 6 months of age. The differences in vertebral shapes and further distinctions between human and sheep spines such as biomechanics, facet anatomy, and rib cage morphology have to be considered when interpreting results after experimental surgery.

  1. Rapid changes in the gut microbiome during human evolution

    OpenAIRE

    Moeller, Andrew H.; Li, Yingying; Mpoudi Ngole, Eitel; Ahuka-Mundeke, Steve; Lonsdorf, Elizabeth V.; Pusey, Anne E.; Peeters, Martine; Hahn, Beatrice H.; Ochman, Howard

    2014-01-01

    Humans are ecosystems containing trillions of microorganisms, but the evolutionary history of this microbiome is obscured by a lack of knowledge about microbiomes of African apes. We sequenced the gut communities of hundreds of chimpanzees, bonobos, and gorillas and developed a phylogenetic approach to reconstruct how present-day human microbiomes have diverged from those of ancestral populations. Compositional change in the microbiome was slow and clock-like during African ape diversificatio...

  2. Human relations with soil are changing rapidly: SSSA's new Work Group on Soil Change

    Science.gov (United States)

    Humanity has rapidly become Earth’s chief agent of soil change, and geologists have named the epoch in which we live the Anthropocene, due to the global scale of human impact on the environment, including soil. In response to the increasing influence of humans on soil processes, the disciplines of ...

  3. Psychiatry and human rights: a difficult relationship, but with a growing potential.

    Science.gov (United States)

    Jarab, Jan

    2015-01-01

    Persons with psychosocial disabilities (mental health problems) are under the protection of the new United Nations Convention on Rights of Persons with Disabilities (CRPD). The CRPD brings a human rights-based approach to disability: it challenges paternalistic views by emphasizing the person as a rights-holder, an active subject, and not just a passive object of care. It also represents a challenge to mainstream human rights movements and mechanisms who have long paid insufficient attention to human rights of persons with (psychosocial) disabilities. It is increasingly understood that human rights of persons with psychosocial disabilities (mental health problems) should not be seen in the narrow perspective, as if the only issue was the most controversial one, that is, deprivation of liberty. In many areas, reform-minded psychiatrists have themselves initiated human rights-friendly reforms. For instance, efforts to implement article 19 of the CRPD—independent living and inclusion in the community—are increasingly becoming part of the mainstream in mental health care. There is potential for further synergy between mental health professionals and human rights activists in looking at the whole range of civil, political, economic, and social rights listed in the Universal Declaration of Human Rights—realizing that all these rights apply also to persons with psychosocial disabilities, and working together towards removing real-life obstacles to their enjoyment.The building of bridges between the two different types of expertise should be encouraged. In this regard, psychiatry would benefit from more cooperation across borders as well as with international human rights bodies, non-governmental organizations and persons with psychosocial disabilities themselves

  4. Pathogen Transmission from Humans to Great Apes is a Growing Threat to Primate Conservation.

    Science.gov (United States)

    Dunay, Emily; Apakupakul, Kathleen; Leard, Stephen; Palmer, Jamie L; Deem, Sharon L

    2018-01-23

    All six great ape species are listed as endangered or critically endangered by the IUCN and experiencing decreasing population trends. One of the threats to these non-human primates is the transmission of pathogens from humans. We conducted a literature review on occurrences of pathogen transmission from humans to great apes to highlight this often underappreciated issue. In total, we found 33 individual occurrences of probable or confirmed pathogen transmission from humans to great apes: 23 involved both pathogen and disease transmission, 7 pathogen transmission only, 2 positive antibody titers to zoonotic pathogens, and 1 pathogen transmission with probable disease. Great ape populations were categorized into captive, semi-free-living, and free-living conditions. The majority of occurrences involved chimpanzees (Pan troglodytes) (n = 23) or mountain gorillas (Gorilla beringei beringei) (n = 8). These findings have implications for conservation efforts and management of endangered great ape populations. Future efforts should focus on monitoring and addressing zoonotic pathogen and disease transmission between humans, great ape species, and other taxa to ensure the health of humans, wild and domestic animals, and the ecosystems we share.

  5. Rapid discrimination of visual scene content in the human brain.

    Science.gov (United States)

    Anokhin, Andrey P; Golosheykin, Simon; Sirevaag, Erik; Kristjansson, Sean; Rohrbaugh, John W; Heath, Andrew C

    2006-06-06

    The rapid evaluation of complex visual environments is critical for an organism's adaptation and survival. Previous studies have shown that emotionally significant visual scenes, both pleasant and unpleasant, elicit a larger late positive wave in the event-related brain potential (ERP) than emotionally neutral pictures. The purpose of the present study was to examine whether neuroelectric responses elicited by complex pictures discriminate between specific, biologically relevant contents of the visual scene and to determine how early in the picture processing this discrimination occurs. Subjects (n = 264) viewed 55 color slides differing in both scene content and emotional significance. No categorical judgments or responses were required. Consistent with previous studies, we found that emotionally arousing pictures, regardless of their content, produce a larger late positive wave than neutral pictures. However, when pictures were further categorized by content, anterior ERP components in a time window between 200 and 600 ms following stimulus onset showed a high selectivity for pictures with erotic content compared to other pictures regardless of their emotional valence (pleasant, neutral, and unpleasant) or emotional arousal. The divergence of ERPs elicited by erotic and non-erotic contents started at 185 ms post-stimulus in the fronto-central midline region, with a later onset in parietal regions. This rapid, selective, and content-specific processing of erotic materials and its dissociation from other pictures (including emotionally positive pictures) suggests the existence of a specialized neural network for prioritized processing of a distinct category of biologically relevant stimuli with high adaptive and evolutionary significance.

  6. Human mobility networks and persistence of rapidly mutating pathogens.

    Science.gov (United States)

    Aleta, Alberto; Hisi, Andreia N S; Meloni, Sandro; Poletto, Chiara; Colizza, Vittoria; Moreno, Yamir

    2017-03-01

    Rapidly mutating pathogens may be able to persist in the population and reach an endemic equilibrium by escaping hosts' acquired immunity. For such diseases, multiple biological, environmental and population-level mechanisms determine the dynamics of the outbreak, including pathogen's epidemiological traits (e.g. transmissibility, infectious period and duration of immunity), seasonality, interaction with other circulating strains and hosts' mixing and spatial fragmentation. Here, we study a susceptible-infected-recovered-susceptible model on a metapopulation where individuals are distributed in sub-populations connected via a network of mobility flows. Through extensive numerical simulations, we explore the phase space of pathogen's persistence and map the dynamical regimes of the pathogen following emergence. Our results show that spatial fragmentation and mobility play a key role in the persistence of the disease whose maximum is reached at intermediate mobility values. We describe the occurrence of different phenomena including local extinction and emergence of epidemic waves, and assess the conditions for large-scale spreading. Findings are highlighted in reference to previous studies and to real scenarios. Our work uncovers the crucial role of hosts' mobility on the ecological dynamics of rapidly mutating pathogens, opening the path for further studies on disease ecology in the presence of a complex and heterogeneous environment.

  7. Simple latex agglutination assay for rapid serodiagnosis of human leptospirosis

    NARCIS (Netherlands)

    Smits, H. L.; van der Hoorn, M. A.; Goris, M. G.; Gussenhoven, G. C.; Yersin, C.; Sasaki, D. M.; Terpstra, W. J.; Hartskeerl, R. A.

    2000-01-01

    A newly developed latex agglutination assay for the detection of genus-specific Leptospira antibodies in human sera was evaluated. The assay is performed by mixing, on an agglutination card, serum with equal volumes of stabilized antigen-coated, dyed test and control latex beads and is read within 2

  8. Low-density microarray technologies for rapid human norovirus genotyping

    Science.gov (United States)

    Human noroviruses cause up to 21 million cases of foodborne disease in the United States annually and are the most common cause of acute gastroenteritis in industrialized countries. To reduce the burden of foodborne disease associated with viruses, the use of low density DNA microarrays in conjuncti...

  9. Rapid latex agglutination test for the serodiagnosis of human brucellosis

    NARCIS (Netherlands)

    Abdoel, Theresia H.; Smits, Henk L.

    2007-01-01

    We developed and evaluated a user-friendly latex agglutination assay for the serodiagnosis of human brucellosis. The assay was obtained by coating colored latex beads with Brucella lipopolysaccharides and drying of the activated beads onto white agglutination cards. Individual cards were sealed in a

  10. Human mobility networks and persistence of rapidly mutating pathogens

    CERN Document Server

    Aleta, Alberto; Meloni, Sandro; Poletto, Chiara; Colizza, Vittoria; Moreno, Yamir

    2016-01-01

    Rapidly mutating pathogens may be able to persist in the population and reach an endemic equilibrium by escaping hosts' acquired immunity. For such diseases, multiple biological, environmental and population-level mechanisms determine the dynamics of the outbreak, including pathogen's epidemiological traits (e.g. transmissibility, infectious period and duration of immunity), seasonality, interaction with other circulating strains and hosts' mixing and spatial fragmentation. Here, we study a susceptible-infected-recovered-susceptible model on a metapopulation where individuals are distributed in subpopulations connected via a network of mobility flows. Through extensive numerical simulations, we explore the phase space of pathogen's persistence and map the dynamical regimes of the pathogen following emergence. Our results show that spatial fragmentation and mobility play a key role in the persistence of the disease whose maximum is reached at intermediate mobility values. We describe the occurrence of differen...

  11. Artificial sweat composition to grow and sustain a mixed human axillary microbiome.

    Science.gov (United States)

    Callewaert, Chris; Buysschaert, Benjamin; Vossen, Els; Fievez, Veerle; Van de Wiele, Tom; Boon, Nico

    2014-08-01

    A novel artificial sweat composition, Skin Community Interaction simulation, designed to mimic the human axillary sweat, was compared to other artificial sweat compositions. Axillary microbiota grown in the novel composition closely resembled the original community. Volatile organic compound analysis showed good correlations with in vivo axillary (mal)odor components. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Etiology and Rapid Diagnosis of Human Viral Gastroenteritis.

    Science.gov (United States)

    1987-05-01

    immunity to rotavirus are complex (1). It seems likely that serum antibody to the virus is associated with protection from illness, and perhaps, local ...three) subgroups of the virus (1). Immune responses are heteroserotypic and heterosubgroup in nature, and various human and animal rotaviruses are...illness typically produces severe diarrhea that commonly lasts for five to eight days and is usually accompanied by fever and vomiting. Rotavirus , which

  13. Evaluation of capillary and myofiber density in the pectoralis major muscles of rapidly growing, high-yield broiler chickens during increased heat stress.

    Science.gov (United States)

    Joiner, K S; Hamlin, G A; Lien, A R J; Bilgili, S F

    2014-09-01

    Skeletal muscle development proceeds from early embryogenesis through marketing age in broiler chickens. While myofiber formation is essentially complete at hatching, myofiber hypertrophy can increase after hatch by assimilation of satellite cell nuclei into myofibers. As the diameter of the myofibers increases, capillary density peripheral to the myofiber is marginalized, limiting oxygen supply and subsequent diffusion into the myofiber, inducing microischemia. The superficial and deep pectoralis muscles constitute 25% of the total body weight in a market-age bird; thus compromise of those muscle groups can have profound economic impact on broiler production. We hypothesized that marginal capillary support relative to the hypertrophic myofibers increases the incidence of microischemia, especially in contemporary high-yield broilers under stressing conditions such as high environmental temperatures. We evaluated the following parameters in four different broiler strains at 39 and 53 days of age when reared under thermoneutral (20 to 25 C) versus hot (30 to 35 C) environmental conditions: capillary density, myofiber density and diameter, and degree of myodegeneration. Our data demonstrate that myofiber diameter significantly increased with age (P > or = 0.0001), while the absolute numbers of capillaries, blood vessels, and myofibers visible in five 400 x microscopic fields decreased (P > or = 0.0001). This is concomitant with marginalization of vascular support in rapidly growing myofibers. The myofiber diameter was significantly lower with hot environmental temperatures (P > or = 0.001); therefore, the absolute number of myofibers visible in five 400X microscopic fields was significantly higher. The incidence and subjective degree of myodegeneration characterized by loss of cross-striations, myocyte hyperrefractility, sarcoplasmic vacuolation, and nuclear pyknosis or loss also increased in hot conditions. Differences among strains were not observed.

  14. Point-of-use membrane filtration and hyperchlorination to prevent patient exposure to rapidly growing mycobacteria in the potable water supply of a skilled nursing facility.

    Science.gov (United States)

    Williams, Margaret M; Chen, Tai-Ho; Keane, Tim; Toney, Nadege; Toney, Sean; Armbruster, Catherine R; Butler, W Ray; Arduino, Matthew J

    2011-09-01

    Healthcare-associated outbreaks and pseudo-outbreaks of rapidly growing mycobacteria (RGM) are frequently associated with contaminated tap water. A pseudo-outbreak of Mycobacterium chelonae-M. abscessus in patients undergoing bronchoscopy was identified by 2 acute care hospitals. RGM was identified in bronchoscopy specimens of 28 patients, 25 of whom resided in the same skilled nursing facility (SNF). An investigation ruled out bronchoscopy procedures, specimen collection, and scope reprocessing at the hospitals as sources of transmission. To identify the reservoir for RGM within the SNF and evaluate 2 water system treatments, hyperchlorination and point-of-use (POU) membrane filters, to reduce RGM. A comparative in situ study of 2 water system treatments to prevent RGM transmission. An SNF specializing in care of patients requiring ventilator support. RGM and heterotrophic plate count (HPC) bacteria were examined in facility water before and after hyperchlorination and in a subsequent 24-week assessment of filtered water by colony enumeration on Middlebrook and R2A media. Mycobacterium chelonae was consistently isolated from the SNF water supply. Hyperchlorination reduced RGM by 1.5 log(10) initially, but the population returned to original levels within 90 days. Concentration of HPC bacteria also decreased temporarily. RGM were reduced below detection level in filtered water, a 3-log(10) reduction. HPC bacteria were not recovered from newly installed filters, although low quantities were found in water from 2-week-old filters. POU membrane filters may be a feasible prevention measure for healthcare facilities to limit exposure of sensitive individuals to RGM in potable water systems.

  15. Speleothem isotopic evidence for rapid human-induced expansion of grasslands in Madagascar at 890 CE

    Science.gov (United States)

    Burns, S. J.; Godfrey, L.; Faina, P.; McGee, D.; Hardt, B. F.; Ranivoharimanana, L.; Randrianasy, J.

    2015-12-01

    The degree to which human activity impacted the landscape, vegetation and fauna of Madagascar remains under debate. Since the early 1920's, the prevailing hypothesis has been that the savannah grasslands that now cover 70% of Madagascar were the result of deforestation, which has also been tied to the disappearance of much of the island's endemic megafauna. Other studies suggest that Madagascar's grasslands are largely natural and that megafaunal extinctions may be climatically induced, leading some authors to question the entire narrative of extensive alteration of the landscape by early human activity. We collected two stalagmites, M14-AB2 and M14-AB3, from Anjohibe Cave in northwestern Madagascar (15.55°S, 46.89°E, 100 masl). Age models were constructed using 8 U/Th age determinations from AB2 and 10 from AB3. The samples began to grow at ~500 CE and were active at the time of collection. Carbon and oxygen stable isotope ratios were measured on 266 samples from AB2 and 173 samples from AB3, yielding sub-decadal temporal resolution. A rapid, more than 10 per mil increase in stalagmite carbon stable isotope ratios documents an almost complete transformation of the landscape from one with a flora dominated by C3 plants to a C4 grassland system. This transformation, well replicated in both stalagmites, occurred at approximately 890 +/- 20 CE and was complete in 100 years. Further, relatively constant oxygen isotope ratios across the carbon isotope transition demonstrate that landscape alteration was not related to changes in climate. We hypothesize that the transformation was caused primarily by expansion of the use of fire by early inhabitants of Madagascar to promote agriculture and the growth of grass as fodder for cattle. The resulting loss of forest habitat very likely increased environmental pressures on Madagascar's megafauna and accelerated their disappearance.

  16. Rapid, automated mosaicking of the human corneal subbasal nerve plexus.

    Science.gov (United States)

    Vaishnav, Yash J; Rucker, Stuart A; Saharia, Keshav; McNamara, Nancy A

    2017-11-27

    Corneal confocal microscopy (CCM) is an in vivo technique used to study corneal nerve morphology. The largest proportion of nerves innervating the cornea lie within the subbasal nerve plexus, where their morphology is altered by refractive surgery, diabetes and dry eye. The main limitations to clinical use of CCM as a diagnostic tool are the small field of view of CCM images and the lengthy time needed to quantify nerves in collected images. Here, we present a novel, rapid, fully automated technique to mosaic individual CCM images into wide-field maps of corneal nerves. We implemented an OpenCV image stitcher that accounts for corneal deformation and uses feature detection to stitch CCM images into a montage. The method takes 3-5 min to process and stitch 40-100 frames on an Amazon EC2 Micro instance. The speed, automation and ease of use conferred by this technique is the first step toward point of care evaluation of wide-field subbasal plexus (SBP) maps in a clinical setting.

  17. Wildfire smoke exposure and human health: Significant gaps in research for a growing public health issue.

    Science.gov (United States)

    Black, Carolyn; Tesfaigzi, Yohannes; Bassein, Jed A; Miller, Lisa A

    2017-10-01

    Understanding the effect of wildfire smoke exposure on human health represents a unique interdisciplinary challenge to the scientific community. Population health studies indicate that wildfire smoke is a risk to human health and increases the healthcare burden of smoke-impacted areas. However, wildfire smoke composition is complex and dynamic, making characterization and modeling difficult. Furthermore, current efforts to study the effect of wildfire smoke are limited by availability of air quality measures and inconsistent air quality reporting among researchers. To help address these issues, we conducted a substantive review of wildfire smoke effects on population health, wildfire smoke exposure in occupational health, and experimental wood smoke exposure. Our goal was to evaluate the current literature on wildfire smoke and highlight important gaps in research. In particular we emphasize long-term health effects of wildfire smoke, recovery following wildfire smoke exposure, and health consequences of exposure in children. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Sustainability: A Crucial Quest for Humanity - Welcome to a New Open Access Journal for a Growing Multidisciplinary Community

    Directory of Open Access Journals (Sweden)

    Marc A. Rosen

    2009-03-01

    Full Text Available Sustainability, although often hard to define precisely, is a rapidly growing area of study that is becoming increasingly applied in diverse areas. The definition put forth in 1983 by the World Commission on Environment and Development, informally known as the Brundtland Commission, captures many aspects of the topic. That commission defined sustainable development as “development that meets the needs of the present without compromising the ability of future generations to meet their own needs.” Other attempts have been made to define what we mean when we refer to sustainability or strive to achieve it as an objective. Despite the differences in definitions, a key theme that emerges is that sustainability is a concept that needs to be incorporated in many if not all of the activities that people undertake. [...

  19. Rapidly progressive periodontal disease associated with human immunodeficiency virus.

    Science.gov (United States)

    Al-Hezaimi, Khalid; Javed, Fawad; Ali, Tazeen Saeed; Al-Askar, Mansour; Al-Rasheed, Abdulaziz

    2012-03-01

    Severe periodontal inflammation with generalized dental plaque accumulation, spontaneous and severe gingival bleeding, fungal infection, and interdental papillae necrosis are presented in a patient infected with human immunodeficiency virus (HIV). Bite-wing radiographs revealed a generalized horizontal alveolar bone loss of 7-8 millimetres in both arches. Erythematous patches were noted on the gingival mucosa in both jaws. DNA testing was performed to indentify the periodontopathogens. The patient had no signs or symptoms of acquired immunodeficiency syndrome. This case-report presents the massive periodontal destruction that occurred in a patient infected with HIV. Therefore, it is highly recommended that patients infected with HIV should be regularly monitored to aid in early detection and to provide proper management of periodontal inflammatory conditions to minimize its destruction.

  20. 3D virtual human rapid modeling method based on top-down modeling mechanism

    Directory of Open Access Journals (Sweden)

    LI Taotao

    2017-01-01

    Full Text Available Aiming to satisfy the vast custom-made character demand of 3D virtual human and the rapid modeling in the field of 3D virtual reality, a new virtual human top-down rapid modeling method is put for-ward in this paper based on the systematic analysis of the current situation and shortage of the virtual hu-man modeling technology. After the top-level realization of virtual human hierarchical structure frame de-sign, modular expression of the virtual human and parameter design for each module is achieved gradu-al-level downwards. While the relationship of connectors and mapping restraints among different modules is established, the definition of the size and texture parameter is also completed. Standardized process is meanwhile produced to support and adapt the virtual human top-down rapid modeling practice operation. Finally, the modeling application, which takes a Chinese captain character as an example, is carried out to validate the virtual human rapid modeling method based on top-down modeling mechanism. The result demonstrates high modelling efficiency and provides one new concept for 3D virtual human geometric mod-eling and texture modeling.

  1. A retrospective approach to assess human health risks associated with growing air pollution in urbanized area of Thar Desert, western Rajasthan, India.

    Science.gov (United States)

    Rumana, Harcharan Singh; Sharma, Ramesh Chandra; Beniwal, Vikas; Sharma, Anil Kumar

    2014-01-09

    : Air pollution has been a matter of great concern globally because of the associated health risks to individuals. The situation is getting worse in developing countries with more urbanization, industrialization and more importantly the rapidly growing population posing a threat to human life in the form of pulmonary, cardiovascular, carcinogenic or asthmatic diseases by accumulating toxic pollutants, harmful gases, metals, hydrocarbons etc. The present study was undertaken to assess the magnitude of ambient air pollutants and their human health risks like respiratory ailments, infectious diseases, cardiovascular diseases and cancer using a Retrospective Approach of Bart Ostra. The parameters PM2.5, PM10, NOx, SO2, NH3 and O3 were monitored at all selected study sites monitored through a high volume sampler (APM 451 Envirotech, Envirotech Instruments Pvt. Ltd., New Delhi, India). Retrospective Approach was used for assessment of risk factors and disease burden of respiratory and cardiopulmonary health problems. Environmental burden of disease showed that the problem of health related to air pollution is a main concern particularly in the growing cities of India. High to critical level of air pollution including PM10, PM2.5, NOx, SO2, NH3 and O3 was observed in all seasons at traffic intersections and commercial sites. The respiratory infections (25% incidence in population exposed to indoor smoke problems) and a prevalence of asthma/COPD (4.4%) in households exposed to high vehicular pollution along with signs of coronary artery/heart disease and/or hypertension and cancers (37.9-52.2%), were reported requiring preventive measures. The study reflects a great concern for the mankind with the need of having streamline ways to limit air pollution and emphasize upon efficiently determining the risk of illness upon exposure to air pollution.

  2. Estimating human ovarian non-growing follicle number: the application of modern stereology techniques to an old problem.

    Science.gov (United States)

    Charleston, Jay S; Hansen, Karl R; Thyer, Angela C; Charleston, Lynne B; Gougeon, Alain; Siebert, Joseph R; Soules, Michael R; Klein, Nancy A

    2007-08-01

    BACKGROUND Previous published reports on the number of non-growing follicles (NGFs) in the human ovary have employed model-based methods for number estimates. These methods are time-intensive, and require correction factors and assumptions that ultimately limit their accuracy. Here, we describe the modification, application and validation of a modern fractionator/optical disector technique for the estimation of human ovarian NGF number. METHODS Forty-eight pairs of normal human ovaries were collected from women (age 8-51 years) undergoing elective bilateral oophorectomy, organ donation, or from autopsy. After gross pathologic examination, systematic random sampling was utilized to obtain tissue for analysis by the fractionator/optical disector method. The precision of individual NGF counts was determined by calculating the observed coefficient of error (OCE). Intra-observer variability and variation in NGF number between ovaries within a pair were also determined. RESULTS The mean OCE was 16.6% with larger variations observed at lower follicle counts. In recount experiments of the same ovary, NGF number estimates varied by 15-29%, except at very low follicle counts where variation was greater, but absolute differences were small. There was no significant difference in NGF number between ovaries within a pair (Wilcoxon signed rank test, P = 0.81). CONCLUSIONS Modern stereology methods provide an unbiased, efficient method for estimating NGF number in the human ovary. Both ovaries within a pair contain similar numbers of NGFs.

  3. Humanizing of process of education of growing personality: morality and its influence on young people, going in for sports

    Directory of Open Access Journals (Sweden)

    Feuerman W. W.

    2012-11-01

    Full Text Available The different aspects of humane education of growing personality are considered. 61 teenagers take part in research (youths. A questionnaire was conducted for the purpose of determination of priority of values teenagers. Questions, touching sporting activity and everyday-domestic relations, were put in a questionnaire. The sociological analysis of relation of personality is conducted to the cultural values of body and physical culture in modern terms. Motive forces of process of mastering of conformities to the law of organization of the system of public relations are exposed. It is marked that the heaved up a problem decides through comparison of biological and social levels of culture of personality. It is set that the regulative and supervisory role of social institutes is broken in modern terms; satisfaction is laboured and the hierarchy of vital necessities is broken.

  4. Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin.

    Science.gov (United States)

    Stoffel, Nicole U; Zeder, Christophe; Fort, Eloïse; Swinkels, Dorine W; Zimmermann, Michael B; Moretti, Diego

    2017-07-26

    Hepcidin is the central systemic regulator of iron metabolism, but its quantification in biological fluids is challenging. Rapid, accurate and user-friendly methods are needed. Our aim was to assess the ability of hepcidin as measured by three different c-ELISA assays to predict iron bioavailability in humans. The three assays used were commercially available DRG and Peninsula assays and the c-ELISA method performed at Radboud University Medical Centre, Nijmegen, The Netherlands (Hepcidinanalysis.com), validated by comparative measurements with time-of-flight mass spectrometry. We analyzed plasma samples (n=37) selected to represent a broad range of hepcidin concentrations from a subgroup of healthy, iron-depleted women in a study assessing fractional absorption from iron supplements. In single regressions, all three c-ELISA assays were predictors of fractional iron absorption: R2=0.363 (DRG), R2=0.281 (Peninsula) and R2=0.327 (Hepcidinanalysis.com). In multiple regressions, models including hepcidin measured with either DRG-, Peninsula or Hepcidinanalysis.com explained 55.7%, 44.5% and 52.5% of variance in fractional absorption, and hepcidin was a strong predictor of fractional absorption irrespective of the hepcidin assays used. However, we found significant differences in absolute values for hepcidin between different methods. Both the DRG assay's (y=0.61x+0.87; R2=0.873) and the Peninsula assay's measurements (y=1.88x+0.62; R2=0.770) were correlated with Hepcidinanalysis.com. The biological variability in plasma hepcidin, (inter-sample CV) was 5-10-fold higher for both the Peninsula and DRG assay than the analytical variably (inter-run within-sample CV) suggesting substantial discriminatory power to distinguish biological hepcidin variation. Between methods, prediction of iron bioavailability in generally healthy iron depleted subjects appears comparable.

  5. Humans and viticulture in Sardinia: The history and social relations as signs of identity of the wine-growing area

    Directory of Open Access Journals (Sweden)

    Benedetto Graziella

    2014-01-01

    Full Text Available The premise of this paper is that viticulture is an expression of history and social relations. In this sense, we embrace a post-modern vision of development that characterized both economic and cultural geography and agricultural economics. Such an approach does consider culture as an element of mediation between humans and the nature, placing it at the heart of the wine-growing territory. So history and social relations have influenced the today spatial densification by types of grape and the persistence, the reduction and/or disappearance of vines’ cultivations due to the different level of integration between humans and wine territories in the Italian region of Sardinia. In this region, there are selected areas where winegrowers have been forced to grub vineyards up, depleting the regional viticultural heritage, others–within which the fabric of the system of social relationships were denser–and where we saw a real rush to purchase of replanting rights for the expansion of the production surface for the increasing of production. The aim of this paper is to highlight the role of history and social relations in the determination of the structure of the regional viticulture through the identification and analysis of diverse case studies.

  6. Rapid development in vitro and in vivo of resistance to ceftazidime in biofilm-growing Pseudomonas aeruginosa due to chromosomal beta-lactamase

    DEFF Research Database (Denmark)

    Bagge, N; Ciofu, O; Skovgaard, L T

    2000-01-01

    The aim of this study was to examine the development of resistance of biofilm-growing P. aeruginosa during treatment with ceftazidime. Biofilms were established in vitro using a modified Robbins device (MRD) and in vivo in the rat model of chronic lung infection. Three P. aeruginosa strains...

  7. Casein kinase II is elevated in solid human tumours and rapidly proliferating non-neoplastic tissue

    DEFF Research Database (Denmark)

    Münstermann, U; Fritz, G; Seitz, G

    1990-01-01

    Protein kinase CKII (i.e. casein kinase II, CKII, NII) is expressed at a higher level in rapidly proliferating tissues and in solid human tumours (e.g. colorectal carcinomas) when compared to the corresponding non-neoplastic colorectal mucosa. This could be shown by (a) Western blotting of cellular...

  8. Rapid detection of human parechoviruses in clinical samples by real-time PCR

    NARCIS (Netherlands)

    Benschop, Kimberley; Molenkamp, Richard; van der Ham, Alwin; Wolthers, Katja; Beld, Marcel

    2008-01-01

    BACKGROUND: Human parechoviruses (HPeVs) have been associated with severe conditions such as neonatal sepsis and meningitis in young children. Rapid identification of an infectious agent in such serious conditions in these patients is essential for adequate decision making regarding treatment and

  9. Human Plasmodium knowlesi infection detected by rapid diagnostic tests for malaria

    NARCIS (Netherlands)

    J.J. van Hellemond (Jaap); M. Rutten (Martine); R. Koelewijn (Rob); A.M. Zeeman (Anne Marie); J. Verweij (Jaap); P.J. Wismans (Pieter); C.H. Kocken (Clemens); P.J.J. van Genderen (Perry)

    2009-01-01

    textabstractWe describe a PCR-confirmed case of Plasmodium knowlesi infection with a high parasitemia level and clinical signs of severe malaria in a migrant worker from Malaysian Borneo in the Netherlands. Investigations showed that commercially available rapid antigen tests for detection of human

  10. Identification of different subtypes of rapid growing Atypical Mycobacterium from water and soil sources: Using PCR-RFLP using hsp65 and rRNA 16s-23s genes.

    Science.gov (United States)

    Varahram, Mohammad; Farnia, Parissa; Saif, Shima; Marashian, Mehran; Farnia, Poopak; Ghanavi, Jaladein; Velayati, Ali Akbar

    2016-12-01

    Nontuberculosis mycobacteria (NTM) are a diverse group of microorganisms that cause a variety of diseases in humans including skin, respiratory, and gastrointestinal tract infection. Generally, NTM are classified into two categories: rapid (7days). In this study, we aimed to investigate NTM frequency and prevalence in environmental samples. Additionally, we tried to identify various subtypes of isolated rapid growing mycobacteria (RGM). Through a prospective descriptive cross-sectional study, water and soil samples were gathered from four neighboring towns around Tehran, the capital of Iran, at different geographic directions. Every 100m(2) of the studied areas gave one sample containing 6g of soil in 3-5cm depth deposited in 50mL sterile water as sampling media. After digestion and decontamination, DNA from culture-positive specimens (RGM) were extracted using phenol-chloroform methods. Then the molecular identification of species and subspecies were performed using 16s-23s rRNA and hsp65 gene. In total, 341 RGM were found, out of which 322 (94.4%) were identified and 20 (5.8%) could not be identified. The most frequent RGM was, Mycobacterium fortuitum (72; 22%), Mycobacterium senegalense (58; 17.7%), Mycobacterium parafortuitum (44; 13.4%) and Mycobacterium conceptionense type 1 (24; 7.2%), and Mycobacterium cheloni type 1 (20; 6.0%). As shown in Table 1, M. fortuitum had more subtypes (8), and the frequency of subtypes 1 (27.7%), 4 (16.6%), and 5 (13.8%) were higher. Among subtypes of M. senegalense, subtype 1 had a higher frequency (70.4%) in comparison to subtype 2 (29.5%). M. cheloni had just one subtype. Our results showed M. fortuitum as the most prominent strain isolated from environmental samples. The frequency was similar in different places, irrespective of climatic variations. Availability of various subtypes of M. fortuitum might indicate a large circulation of this RGM in soil and water of Iranian territory. This high prevalence of M. fortuitum might

  11. Rapid sensing of l-leucine by human and murine hypothalamic neurons: Neurochemical and mechanistic insights.

    Science.gov (United States)

    Heeley, Nicholas; Kirwan, Peter; Darwish, Tamana; Arnaud, Marion; Evans, Mark L; Merkle, Florian T; Reimann, Frank; Gribble, Fiona M; Blouet, Clemence

    2018-02-07

    Dietary proteins are sensed by hypothalamic neurons and strongly influence multiple aspects of metabolic health, including appetite, weight gain, and adiposity. However, little is known about the mechanisms by which hypothalamic neural circuits controlling behavior and metabolism sense protein availability. The aim of this study is to characterize how neurons from the mediobasal hypothalamus respond to a signal of protein availability: the amino acid l-leucine. We used primary cultures of post-weaning murine mediobasal hypothalamic neurons, hypothalamic neurons derived from human induced pluripotent stem cells, and calcium imaging to characterize rapid neuronal responses to physiological changes in extracellular l-Leucine concentration. A neurochemically diverse subset of both mouse and human hypothalamic neurons responded rapidly to l-leucine. Consistent with l-leucine's anorexigenic role, we found that 25% of mouse MBH POMC neurons were activated by l-leucine. 10% of MBH NPY neurons were inhibited by l-leucine, and leucine rapidly reduced AGRP secretion, providing a mechanism for the rapid leucine-induced inhibition of foraging behavior in rodents. Surprisingly, none of the candidate mechanisms previously implicated in hypothalamic leucine sensing (K ATP channels, mTORC1 signaling, amino-acid decarboxylation) were involved in the acute activity changes produced by l-leucine. Instead, our data indicate that leucine-induced neuronal activation involves a plasma membrane Ca 2+ channel, whereas leucine-induced neuronal inhibition is mediated by inhibition of a store-operated Ca 2+ current. A subset of neurons in the mediobasal hypothalamus rapidly respond to physiological changes in extracellular leucine concentration. Leucine can produce both increases and decreases in neuronal Ca 2+ concentrations in a neurochemically-diverse group of neurons, including some POMC and NPY/AGRP neurons. Our data reveal that leucine can signal through novel mechanisms to rapidly

  12. Apnea-Induced Rapid Eye Movement Sleep Disruption Impairs Human Spatial Navigational Memory

    OpenAIRE

    Varga, Andrew W.; Kishi, Akifumi; Mantua, Janna; Lim, Jason; Koushyk, Viachaslau; Leibert, David P.; Osorio, Ricardo S.; Rapoport, David M.; Ayappa, Indu

    2014-01-01

    Hippocampal electrophysiology and behavioral evidence support a role for sleep in spatial navigational memory, but the role of particular sleep stages is less clear. Although rodent models suggest the importance of rapid eye movement (REM) sleep in spatial navigational memory, a similar role for REM sleep has never been examined in humans. We recruited subjects with severe obstructive sleep apnea (OSA) who were well treated and adherent with continuous positive airway pressure (CPAP). Restric...

  13. Rapid functional analysis of computationally complex rare human IRF6 gene variants using a novel zebrafish model.

    Directory of Open Access Journals (Sweden)

    Edward B Li

    2017-09-01

    Full Text Available Large-scale sequencing efforts have captured a rapidly growing catalogue of genetic variations. However, the accurate establishment of gene variant pathogenicity remains a central challenge in translating personal genomics information to clinical decisions. Interferon Regulatory Factor 6 (IRF6 gene variants are significant genetic contributors to orofacial clefts. Although approximately three hundred IRF6 gene variants have been documented, their effects on protein functions remain difficult to interpret. Here, we demonstrate the protein functions of human IRF6 missense gene variants could be rapidly assessed in detail by their abilities to rescue the irf6 -/- phenotype in zebrafish through variant mRNA microinjections at the one-cell stage. The results revealed many missense variants previously predicted by traditional statistical and computational tools to be loss-of-function and pathogenic retained partial or full protein function and rescued the zebrafish irf6 -/- periderm rupture phenotype. Through mRNA dosage titration and analysis of the Exome Aggregation Consortium (ExAC database, IRF6 missense variants were grouped by their abilities to rescue at various dosages into three functional categories: wild type function, reduced function, and complete loss-of-function. This sensitive and specific biological assay was able to address the nuanced functional significances of IRF6 missense gene variants and overcome many limitations faced by current statistical and computational tools in assigning variant protein function and pathogenicity. Furthermore, it unlocked the possibility for characterizing yet undiscovered human IRF6 missense gene variants from orofacial cleft patients, and illustrated a generalizable functional genomics paradigm in personalized medicine.

  14. Modulators of mercury risk to wildlife and humans in the context of rapid global change

    Science.gov (United States)

    Eagles-Smith, Collin A.; Silbergeld, Ellen K.; Basu, Niladri; Bustamante, Paco; Diaz-Barriga, Fernando; Hopkins, William A.; Kidd, Karen A.; Nyland, Jennifer F.

    2018-01-01

    Environmental mercury (Hg) contamination is an urgent global health threat. The complexity of Hg in the environment can hinder accurate determination of ecological and human health risks, particularly within the context of the rapid global changes that are altering many ecological processes, socioeconomic patterns, and other factors like infectious disease incidence, which can affect Hg exposures and health outcomes. However, the success of global Hg-reduction efforts depends on accurate assessments of their effectiveness in reducing health risks. In this paper, we examine the role that key extrinsic and intrinsic drivers play on several aspects of Hg risk to humans and organisms in the environment. We do so within three key domains of ecological and human health risk. First, we examine how extrinsic global change drivers influence pathways of Hg bioaccumulation and biomagnification through food webs. Next, we describe how extrinsic socioeconomic drivers at a global scale, and intrinsic individual-level drivers, influence human Hg exposure. Finally, we address how the adverse health effects of Hg in humans and wildlife are modulated by a range of extrinsic and intrinsic drivers within the context of rapid global change. Incorporating components of these three domains into research and monitoring will facilitate a more holistic understanding of how ecological and societal drivers interact to influence Hg health risks.

  15. Rapid evolution of the cerebellum in humans and other great apes.

    Science.gov (United States)

    Barton, Robert A; Venditti, Chris

    2014-10-20

    Humans' unique cognitive abilities are usually attributed to a greatly expanded neocortex, which has been described as "the crowning achievement of evolution and the biological substrate of human mental prowess". The human cerebellum, however, contains four times more neurons than the neocortex and is attracting increasing attention for its wide range of cognitive functions. Using a method for detecting evolutionary rate changes along the branches of phylogenetic trees, we show that the cerebellum underwent rapid size increase throughout the evolution of apes, including humans, expanding significantly faster than predicted by the change in neocortex size. As a result, humans and other apes deviated significantly from the general evolutionary trend for neocortex and cerebellum to change in tandem, having significantly larger cerebella relative to neocortex size than other anthropoid primates. These results suggest that cerebellar specialization was a far more important component of human brain evolution than hitherto recognized and that technical intelligence was likely to have been at least as important as social intelligence in human cognitive evolution. Given the role of the cerebellum in sensory-motor control and in learning complex action sequences, cerebellar specialization is likely to have underpinned the evolution of humans' advanced technological capacities, which in turn may have been a preadaptation for language. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Rapid spread of mouse mammary tumor virus in cultured human breast cells

    Directory of Open Access Journals (Sweden)

    Günzburg Walter H

    2007-10-01

    Full Text Available Abstract Background The role of mouse mammary tumor virus (MMTV as a causative agent in human breast carcinogenesis has recently been the subject of renewed interest. The proposed model is based on the detection of MMTV sequences in human breast cancer but not in healthy breast tissue. One of the main drawbacks to this model, however, was that until now human cells had not been demonstrated to sustain productive MMTV infection. Results Here, we show for the first time the rapid spread of mouse mammary tumor virus, MMTV(GR, in cultured human mammary cells (Hs578T, ultimately leading to the infection of every cell in culture. The replication of the virus was monitored by quantitative PCR, quantitative RT-PCR and immunofluorescence imaging. The infected human cells expressed, upon cultivation with dexamethasone, MMTV structural proteins and released spiked B-type virions, the infectivity of which could be neutralized by anti-MMTV antibody. Replication of the virus was efficiently blocked by an inhibitor of reverse transcription, 3'-azido-3'-deoxythymidine. The human origin of the infected cells was confirmed by determining a number of integration sites hosting the provirus, which were unequivocally identified as human sequences. Conclusion Taken together, our results show that human cells can support replication of mouse mammary tumor virus.

  17. Management of skeletal Class III malocclusion with unilateral crossbite on a growing patient using facemask-bonded rapid palatal expander and fixed appliances

    Directory of Open Access Journals (Sweden)

    Tinnie Effendy

    2015-01-01

    Full Text Available Facemask (FM and bonded rapid palatal expander (RPE are part of growth modification treatments for correcting skeletal Class III pattern with retrognathic maxilla. This orthopaedic treatment is usually preceded by fixed appliances to achieve aesthetic dental alignment and improve interdigitation. This case report reviews treatment of Class III malocclusion with unilateral crossbite in a 12-year-old boy using FM and bonded RPE, followed by fixed appliances. Choice of FM and bonded RPE was in line with indication which was mild Class III malocclusion with retrognathic maxilla. Execution of treatment was made considering treatment biomechanics and patient cooperation. This orthopaedic treatment was followed by orthodontic treatment specifically aimed to correct unilateral crossbite, canine relationship yet to reach Class I, lower midline shift, as well as unintended dental consequences of using bonded RPE, namely posterior open bite and deepening curve of spee. Posttreatment facial profile and smile are more esthetic. Occlusion is significantly improved both functionally and aesthetically.

  18. Polymer based biosensor for rapid electrochemical detection of virus infection of human cells

    DEFF Research Database (Denmark)

    Kiilerich-Pedersen, Katrine; Poulsen, Claus R.; Jain, Titoo

    2011-01-01

    The demand in the field of medical diagnostics for simple, cost efficient and disposable devices is growing. Here, we present a label free, all-polymer electrochemical biosensor for detection of acute viral disease. The dynamics of a viral infection in human cell culture was investigated in a micro...... fluidic system on conductive polymer PEDOT:TsO microelectrodes by electrochemical impedance spectroscopy and video time lapse microscopy.Employing this sensitive, real time electrochemical technique, we could measure the immediate cell response to cytomegalovirus, and detect an infection within 3h, which...

  19. Development of an in vitro Assay, Based on the BioFilm Ring Test®, for Rapid Profiling of Biofilm-Growing Bacteria.

    Science.gov (United States)

    Di Domenico, Enea G; Toma, Luigi; Provot, Christian; Ascenzioni, Fiorentina; Sperduti, Isabella; Prignano, Grazia; Gallo, Maria T; Pimpinelli, Fulvia; Bordignon, Valentina; Bernardi, Thierry; Ensoli, Fabrizio

    2016-01-01

    Microbial biofilm represents a major virulence factor associated with chronic and recurrent infections. Pathogenic bacteria embedded in biofilms are highly resistant to environmental and chemical agents, including antibiotics and therefore difficult to eradicate. Thus, reliable tests to assess biofilm formation by bacterial strains as well as the impact of chemicals or antibiotics on biofilm formation represent desirable tools for a most effective therapeutic management and microbiological risk control. Current methods to evaluate biofilm formation are usually time-consuming, costly, and hardly applicable in the clinical setting. The aim of the present study was to develop and assess a simple and reliable in vitro procedure for the characterization of biofilm-producing bacterial strains for future clinical applications based on the BioFilm Ring Test® (BRT) technology. The procedure developed for clinical testing (cBRT) can provide an accurate and timely (5 h) measurement of biofilm formation for the most common pathogenic bacteria seen in clinical practice. The results gathered by the cBRT assay were in agreement with the traditional crystal violet (CV) staining test, according to the κ coefficient test (κ = 0.623). However, the cBRT assay showed higher levels of specificity (92.2%) and accuracy (88.1%) as compared to CV. The results indicate that this procedure offers an easy, rapid and robust assay to test microbial biofilm and a promising tool for clinical microbiology.

  20. Community-based rapid oral human immunodeficiency virus testing for tuberculosis patients in Lima, Peru.

    Science.gov (United States)

    Nelson, Adrianne K; Caldas, Adolfo; Sebastian, Jose Luis; Muñoz, Maribel; Bonilla, Cesear; Yamanija, Jose; Jave, Oswaldo; Magan, Christina; Saldivar, Judith; Espiritu, Betty; Rosell, Gustavo; Bayona, Jaime; Shin, Sonya

    2012-09-01

    Among tuberculosis patients, timely diagnosis of human immunodeficiency virus (HIV) co-infection and early antiretroviral treatment are crucial, but are hampered by a myriad of individual and structural barriers. Community-based models to provide counseling and rapid HIV testing are few but offer promise. During November 2009-April 2010, community health workers offered and performed HIV counseling and testing by using the OraQuick Rapid HIV-1/2 Antibody Test to new tuberculosis cases in 22 Ministry of Health establishments and their household contacts (n = 130) in Lima, Peru. Refusal of HIV testing or study participation was low (4.7%). Intervention strengths included community-based approach with participant preference for testing site, use of a rapid, non-invasive test, and accompaniment to facilitate HIV care and family disclosure. We will expand the intervention under programmatic auspices for rapid community-based testing for new tuberculosis cases in high incidence establishments. Other potential target populations include contacts of HIV-positive persons and pregnant women.

  1. Rapid detection and subtyping of human influenza A viruses and reassortants by pyrosequencing.

    Directory of Open Access Journals (Sweden)

    Yi-Mo Deng

    Full Text Available BACKGROUND: Given the continuing co-circulation of the 2009 H1N1 pandemic influenza A viruses with seasonal H3N2 viruses, rapid and reliable detection of newly emerging influenza reassortant viruses is important to enhance our influenza surveillance. METHODOLOGY/PRINCIPAL FINDINGS: A novel pyrosequencing assay was developed for the rapid identification and subtyping of potential human influenza A virus reassortants based on all eight gene segments of the virus. Except for HA and NA genes, one universal set of primers was used to amplify and subtype each of the six internal genes. With this method, all eight gene segments of 57 laboratory isolates and 17 original specimens of seasonal H1N1, H3N2 and 2009 H1N1 pandemic viruses were correctly matched with their corresponding subtypes. In addition, this method was shown to be capable of detecting reassortant viruses by correctly identifying the source of all 8 gene segments from three vaccine production reassortant viruses and three H1N2 viruses. CONCLUSIONS/SIGNIFICANCE: In summary, this pyrosequencing assay is a sensitive and specific procedure for screening large numbers of viruses for reassortment events amongst the commonly circulating human influenza A viruses, which is more rapid and cheaper than using conventional sequencing approaches.

  2. Rapid diagnosis and intraoperative margin assessment of human lung cancer with fluorescence lifetime imaging microscopy

    Directory of Open Access Journals (Sweden)

    Mengyan Wang

    2017-12-01

    Full Text Available A method of rapidly differentiating lung tumor from healthy tissue is extraordinarily needed for both the diagnosis and the intraoperative margin assessment. We assessed the ability of fluorescence lifetime imaging microscopy (FLIM for differentiating human lung cancer and normal tissues with the autofluorescence, and also elucidated the mechanism in tissue studies and cell studies. A 15-patient testing group was used to compare FLIM results with traditional histopathology diagnosis. Based on the endogenous fluorescence lifetimes of the testing group, a criterion line was proposed to distinguish normal and cancerous tissues. Then by blinded examined 41 sections from the validation group of other 16 patients, the sensitivity and specificity of FLIM were determined. The cellular metabolism was studied with specific perturbations of oxidative phosphorylation and glycolysis in cell studies. The fluorescence lifetime of cancerous lung tissues is consistently lower than normal tissues, and this is due to the both decrease of reduced nicotinamide adenine dinucleotide (NADH and flavin adenine dinucleotide (FAD lifetimes. A criterion line of lifetime at 1920 ps can be given for differentiating human lung cancer and normal tissues.The sensitivity and specificity of FLIM for lung cancer diagnosis were determined as 92.9% and 92.3%. These findings suggest that NADH and FAD can be used to rapidly diagnose lung cancer. FLIM is a rapid, accurate and highly sensitive technique in the judgment during lung cancer surgery and it can be potential in earlier cancer detection.

  3. Rapid diagnosis and intraoperative margin assessment of human lung cancer with fluorescence lifetime imaging microscopy.

    Science.gov (United States)

    Wang, Mengyan; Tang, Feng; Pan, Xiaobo; Yao, Longfang; Wang, Xinyi; Jing, Yueyue; Ma, Jiong; Wang, Guifang; Mi, Lan

    2017-12-01

    A method of rapidly differentiating lung tumor from healthy tissue is extraordinarily needed for both the diagnosis and the intraoperative margin assessment. We assessed the ability of fluorescence lifetime imaging microscopy (FLIM) for differentiating human lung cancer and normal tissues with the autofluorescence, and also elucidated the mechanism in tissue studies and cell studies. A 15-patient testing group was used to compare FLIM results with traditional histopathology diagnosis. Based on the endogenous fluorescence lifetimes of the testing group, a criterion line was proposed to distinguish normal and cancerous tissues. Then by blinded examined 41 sections from the validation group of other 16 patients, the sensitivity and specificity of FLIM were determined. The cellular metabolism was studied with specific perturbations of oxidative phosphorylation and glycolysis in cell studies. The fluorescence lifetime of cancerous lung tissues is consistently lower than normal tissues, and this is due to the both decrease of reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) lifetimes. A criterion line of lifetime at 1920 ps can be given for differentiating human lung cancer and normal tissues.The sensitivity and specificity of FLIM for lung cancer diagnosis were determined as 92.9% and 92.3%. These findings suggest that NADH and FAD can be used to rapidly diagnose lung cancer. FLIM is a rapid, accurate and highly sensitive technique in the judgment during lung cancer surgery and it can be potential in earlier cancer detection.

  4. Rapid detection and identification of human hookworm infections through high resolution melting (HRM analysis.

    Directory of Open Access Journals (Sweden)

    Romano Ngui

    Full Text Available BACKGROUND: Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR coupled with high resolution melting-curve (HRM analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species. METHODS: Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2 of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples. CONCLUSION: The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.

  5. Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.

    Science.gov (United States)

    Ngui, Romano; Lim, Yvonne A L; Chua, Kek Heng

    2012-01-01

    Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species. Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples. The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.

  6. Systematic Three-Dimensional Coculture Rapidly Recapitulates Interactions between Human Neurons and Astrocytes

    Directory of Open Access Journals (Sweden)

    Robert Krencik

    2017-12-01

    Full Text Available Summary: Human astrocytes network with neurons in dynamic ways that are still poorly defined. Our ability to model this relationship is hampered by the lack of relevant and convenient tools to recapitulate this complex interaction. To address this barrier, we have devised efficient coculture systems utilizing 3D organoid-like spheres, termed asteroids, containing pre-differentiated human pluripotent stem cell (hPSC-derived astrocytes (hAstros combined with neurons generated from hPSC-derived neural stem cells (hNeurons or directly induced via Neurogenin 2 overexpression (iNeurons. Our systematic methods rapidly produce structurally complex hAstros and synapses in high-density coculture with iNeurons in precise numbers, allowing for improved studies of neural circuit function, disease modeling, and drug screening. We conclude that these bioengineered neural circuit model systems are reliable and scalable tools to accurately study aspects of human astrocyte-neuron functional properties while being easily accessible for cell-type-specific manipulations and observations. : In this article, Krencik and colleagues show that high-density cocultures of pre-differentiated human astrocytes with induced neurons, from pluripotent stem cells, elicit mature characteristics by 3–5 weeks. This provides a faster and more defined alternative method to organoid cultures for investigating human neural circuit function. Keywords: human pluripotent stem cells, neurons, astrocytes, synapses, coculture, three-dimensional spheres, organoids, disease modeling

  7. New anatomical data on the growing C4 vertebra and its three ossification centers in human fetuses.

    Science.gov (United States)

    Baumgart, Mariusz; Szpinda, Michał; Szpinda, Anna

    2013-04-01

    Detailed knowledge on the normative growth of the spine is of great relevance in the prenatal diagnosis of its abnormalities. The present study was conducted to compile age-specific reference data for vertebra C4 and its three ossification centers in human fetuses. With the use of CT (Biograph mCT), digital image analysis (Osirix 3.9) and statistical analysis (Wilcoxon signed-rank test, Kolmogorov-Smirnov test, Levene's test, Student's t test, one-way ANOVA, post hoc RIR Tukey test, linear and nonlinear regression analysis), the normative growth of vertebra C4 and its three ossification centers in 55 spontaneously aborted human fetuses (27 males, 28 females) aged 17-30 weeks was examined. Significant differences in neither sex nor laterality were found. The height and transverse and sagittal diameters of the C4 vertebral body increased logarithmically as: y = -3.866 + 2.225 × ln(Age) ± 0.238 (R(2) = 0.69), y = -7.077 + 3.547 × ln(Age) ± 0.356 (R(2) = 0.72) and y = -3.886 + 2.272 × ln(Age) ± 0.222 (R(2) = 0.73), respectively. The C4 vertebral body grew linearly in cross-sectional area as y = -7.205 + 0.812 × Age ± 1.668 (R(2) = 0.76) and four-degree polynomially in volume as y = 14.108 + 0.00007 × Age(4) ± 6.289 (R(2) = 0.83). The transverse and sagittal diameters, cross-sectional area and volume of the ossification center of the C4 vertebral body generated the following functions: y = -8.836 + 3.708 × ln(Age) ± 0.334 (R(2) = 0.76), y = -7.748 + 3.240 × ln(Age) ± 0.237 (R(2) = 0.83), y = -4.690 + 0.437 × Age ± 1.172 (R(2) = 0.63) and y = -5.917 + 0.582 × Age ± 1.157 (R(2) = 0.77), respectively. The ossification center-to-vertebral body volume ratio gradually declined with age. On the right and left, the neural ossification centers showed the following growth: y = -19.601 + 8.018 × ln(Age) ± 0.369 (R(2) = 0.92) and y = -15.804 + 6.912 × ln(Age) ± 0.471 (R (2) = 0.85) for length, y = -5.806 + 2.587 × ln(Age) ± 0.146 (R(2) = 0.88) and y = -5

  8. Rapid Water Uptake and Limited Storage Capacity at Height of Growing Season in Four Temperate Tree Species in a Central Pennsylvania Catchment

    Science.gov (United States)

    Gaines, K.; Meinzer, F. C.; Duffy, C.; Thomas, E.; Eissenstat, D. M.

    2014-12-01

    rapid water uptake and tree water storage limited to about a month in duration. These findings are necessary for modeling of hydrologic parameters that are influenced by tree water age. They also indicate that trees on shallow soil in this catchment may be at risk if droughts lasting over a month occur more frequently in future years.

  9. Detection and monitoring of human bocavirus 1 infection by a new rapid antigen test.

    Science.gov (United States)

    Bruning, A H L; Susi, P; Toivola, H; Christensen, A; Söderlund-Venermo, M; Hedman, K; Aatola, H; Zvirbliene, A; Koskinen, J O

    2016-05-01

    Clinically relevant diagnosis of human bocavirus 1 (HBoV1) is challenging, as the virus is frequently detected in asymptomatic patients, and cofindings with other respiratory viruses are common. The clinical value of current diagnostic methods, such as PCR, is therefore low, and alternative diagnostic strategies are needed. We describe for the first time the use of an antigen detection assay for the rapid identification of HBoV1 in a paediatric patient with respiratory tract infection symptoms. We estimate the duration of active HBoV1 infection to be 6 days.

  10. Detection and monitoring of human bocavirus 1 infection by a new rapid antigen test

    Directory of Open Access Journals (Sweden)

    A.H.L. Bruning

    2016-05-01

    Full Text Available Clinically relevant diagnosis of human bocavirus 1 (HBoV1 is challenging, as the virus is frequently detected in asymptomatic patients, and cofindings with other respiratory viruses are common. The clinical value of current diagnostic methods, such as PCR, is therefore low, and alternative diagnostic strategies are needed. We describe for the first time the use of an antigen detection assay for the rapid identification of HBoV1 in a paediatric patient with respiratory tract infection symptoms. We estimate the duration of active HBoV1 infection to be 6 days.

  11. Novel patterns for the growing main bronchi in the human fetus: an anatomical, digital and statistical study.

    Science.gov (United States)

    Szpinda, Michał; Daroszewski, Marcin; Woźniak, Alina; Szpinda, Anna; Flisiński, Piotr; Dombek, Małgorzata; Mila-Kierzenkowska, Celestyna; Siedlaczek, Waldemar

    2014-01-01

    Intensive progress in prenatal medicine results in performing airway management in the fetus affected by life-threatening congenital malformations. This study aimed to examine age-specific reference intervals and growth dynamics for length, proximal and distal external transverse diameters, and projection surface areas of the two main bronchi at varying gestational ages, including their relative growth in length and projection surface area. Using anatomical dissection, digital image analysis and statistics, length, proximal and distal external transverse diameters, and projection surface areas of the right and left main bronchi were examined in 73 human fetuses (39 males, 34 females) aged 14-25 weeks, derived from spontaneous abortions and stillbirths. Statistical analysis showed no sex differences. Between the 14 and 25th week of gestation, the lengths of the right and left main bronchi increased from 1.43 ± 0.18 to 3.18 ± 0.39 mm, and from 2.97 ± 0.16 to 7.58 ± 1.95 mm, in accordance with the functions: [Formula: see text], respectively. The proximal external transverse diameters of the right and left main bronchi varied from 2.13 ± 0.41 to 4.24 ± 0.20 mm, and from 1.84 ± 0.06 to 3.67 ± 0.66 mm, following the logarithmic models: [Formula: see text], respectively. The distal external transverse diameter rose from 2.09 ± 0.47 to 4.24 ± 0.20 mm, as [Formula: see text] for the right main bronchus, and from 1.85 ± 0.04 to 3.67 ± 0.66 mm, like [Formula: see text] for the left one. On either side, there were no statistically significant differences between values of the proximal and distal transverse diameters of the main bronchus. The projection surface areas of the right and left main bronchi ranged from 2.95 ± 0.19 to 13.34 ± 2.12 mm(2), and from 5.57 ± 0.21 to 28.52 ± 5.24 mm(2), as [Formula: see text] and [Formula: see text]. The two main bronchi revealed a proportionate increase in both length and projection

  12. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples.

    Science.gov (United States)

    Mirnejad, Reza; Mohamadi, Mozafar; Piranfar, Vahbeh; Mortazavi, Seied Mojtaba; Kachuei, Reza

    2013-06-01

    To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus 39 (B. abortus 39) (23%), 13 for Brucella melitensis 39 (B. melitensis 39) (25%) and 0 for Brucella ovis 39 (B. ovis 39) (0%). This work demonstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  13. Activation of intracellular angiotensin AT2 receptors induces rapid cell death in human uterine leiomyosarcoma cells

    DEFF Research Database (Denmark)

    Zhao, Yi; Lützen, Ulf; Fritsch, Jürgen

    2015-01-01

    of apoptosis and cell death in cultured human uterine leiomyosarcoma (SK-UT-1) cells and control human uterine smooth muscle cells (HutSMC). The intracellular levels of the AT2 receptor are low in proliferating SK-UT-1 cells but the receptor is substantially up-regulated in quiescent SK-UT-1 cells with high...... densities in mitochondria. Activation of the cell membrane AT2 receptors by a concomitant treatment with angiotensin II and the AT1 receptor antagonist, losartan, induces apoptosis but does not affect the rate of cell death. We demonstrate for the first time that the high-affinity, non-peptide AT2 receptor...... agonist, Compound 21 (C21) penetrates the cell membrane of quiescent SK-UT-1 cells, activates intracellular AT2 receptors and induces rapid cell death; approximately 70% of cells died within 24 h. The cells, which escaped from the cell death, displayed activation of the mitochondrial apoptotic pathway, i...

  14. Ion Torren Semiconductor Sequencing Allows Rapid, Low Cost Sequencing of the Human Exome ( 7th Annual SFAF Meeting, 2012)

    Energy Technology Data Exchange (ETDEWEB)

    Jenkins, David [EdgeBio

    2012-06-01

    David Jenkins on "Ion Torrent semiconductor sequencing allows rapid, low-cost sequencing of the human exome" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

  15. Replication of human tracheobronchial hollow airway models using a selective laser sintering rapid prototyping technique.

    Science.gov (United States)

    Clinkenbeard, Rodney E; Johnson, David L; Parthasarathy, Ramkumar; Altan, M Cengiz; Tan, Kah-Hoe; Park, Seok-Min; Crawford, Richard H

    2002-01-01

    Exposures to toxic or pathogenic aerosols are known to produce adverse health effects. The nature and severity of these effects often are governed in large part by the location and amount of aerosol deposition within the respiratory tract. Morphologically detailed replica hollow lung airway casts are widely used in aerosol deposition research; however, techniques are not currently available that allow replicate deposition studies in identical morphologically detailed casts produced from a common reference anatomy. This project developed a technique for the precision manufacture of morphologically detailed human tracheobronchial airway models based on high-resolution anatomical imaging data. Detailed physical models were produced using the selective laser sintering (SLS) rapid prototyping process. Input to the SLS process was a three-dimensional computer model developed by boundary-based two-dimension to three-dimension conversion of anatomical images from the original National Institutes of Health/National Library of Medicine Visible Human male data set. The SLS process produced identical replicate models that corresponded exactly to the anatomical section images, within the limits of the measurement. At least five airway generations were achievable, corresponding to airways less than 2 mm in diameter. It is anticipated that rapid prototyping manufacture of respiratory tract structures based on reference anatomies such as the Visible Male and Visible Female may provide "gold standard" models for inhaled aerosol deposition studies. Adaptations of the models to represent various disease states may be readily achieved, thereby promoting exploration of pharmaceutical research on targeted drug delivery via inhaled aerosols.

  16. Rapid analysis of acetone in human plasma by derivatization desorption electrospray ionization.

    Science.gov (United States)

    Xu, Guangming; Chen, Bo; Liu, Guozhu; Yao, Shouzhuo

    2010-09-01

    Desorption electrospray ionization mass spectrometry (DESI MS) is rapidly becoming a powerful analysis tool for a wide variety of samples in the open air. By combining DESI MS and derivatization, acetone in human blood samples, one of the diabetes mellitus (DM) biomarkers, was rapidly analyzed. Prior to DESI, the plasma sample was first exposed to hydroxylamine hydrochloride and acetone in plasma was derived into fixed oxime. The reactive products were validated by DESI/MS/MS. Quantification was performed in the positive-ion electrospray ionization mode by monitoring of the product ions at m/z 74 for acetone oxime and m/z 116 for 4-methyl-2-pentanone oxime as the internal standard. The limit of detection was of the order of magnitude of 0.10 ppbv and the linear range comprised two decades of concentration, covering the relevant concentration range of acetone in plasma samples. The method has achieved fast, high-throughput analysis of acetone in human plasma with little beforehand preparation and an experiment time of only a few seconds.

  17. Portable platform for rapid in-field identification of human fecal pollution in water.

    Science.gov (United States)

    Jiang, Yu Sherry; Riedel, Timothy E; Popoola, Jessica A; Morrow, Barrett R; Cai, Sheng; Ellington, Andrew D; Bhadra, Sanchita

    2017-12-13

    Human fecal contamination of water is a public health risk. However, inadequate testing solutions frustrate timely, actionable monitoring. Bacterial culture-based methods are simple but typically cannot distinguish fecal host source. PCR assays can identify host sources but require expertise and infrastructure. To bridge this gap we have developed a field-ready nucleic acid diagnostic platform and rapid sample preparation methods that enable on-site identification of human fecal contamination within 80 min of sampling. Our platform relies on loop-mediated isothermal amplification (LAMP) of human-associated Bacteroides HF183 genetic markers from crude samples. Oligonucleotide strand exchange (OSD) probes reduce false positives by sequence specifically transducing LAMP amplicons into visible fluorescence that can be photographed by unmodified smartphones. Our assay can detect as few as 17 copies/ml of human-associated HF183 targets in sewage-contaminated water without cross-reaction with canine or feline feces. It performs robustly with a variety of environmental water sources and with raw sewage. We have also developed lyophilized assays and inexpensive 3D-printed devices to minimize cost and facilitate field application. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. People On The Move: Some Thoughts On Human Dispersal In Relation To Rapid Climatic Change

    Science.gov (United States)

    Davies, W.

    It is still generally assumed that the default situation for past humans must have been to be sedentary. That is to say, given a chance people would have settled in one area (with a good supply of resources) and established clearly-defined territories. Such concepts presuppose that much of human existence was conducted in climatic conditions sim- ilar to the relatively stable ones seen in the Holocene. What effects do rapid climatic fluctuations have upon environmental carrying capacity, and thus upon human mobil- ity and exploitation patterns? Such an approach could be called 'non-analogue', as it does not seek to impose [current] Holocene patterns upon the Pleistocene, in the same way that 'non-analogue' animal and plant communities are now routinely described for the same period. If one adopts non-analogue perspectives, perhaps one could also argue that in many cases mobility was the rule and not the exception. Turning the conventional wisdom around, we can ask why people should remain in an area. What are the characteristics of that area which could have encouraged people to become less mobile? I do not argue that all groups were mobile: some cannot have been, and not every member of other groups would have been equally mobile (differentiation on grounds of age and sex). In addition, mobility patterns must also have varied over time, although we should not necessarily expect a discernible linear trend either towards or away from greater mobility, because such behaviour operates within a climatic and environmental framework as well as a socio-economic one. If climate oscillated rapidly, it is feasible to suggest that such fluctuations affected environmental stability and thus carrying capacity. The resource species present and their availability would therefore affect the possibilities for human mobility. When discussing the possibilities for human dispersal into new regions, we essentially have a choice between two competing models: the Wave of Advance (sensu

  19. A regional human services authority's rapid needs assessment of evacuees following natural disasters.

    Science.gov (United States)

    Post, David E; Kasofsky, Jan M; Hunte, Christopher N; Diaz, James H

    2008-01-01

    The Atlantic hurricane season of 2005 was not an ordinary season, and Hurricane Katrina was not an ordinary hurricane. Hurricane Katrina damaged more than 93,000 square miles of Gulf of Mexico coastline, displaced more than 1 million residents from New Orleans, and flooded more than 80 percent of New Orleans for weeks, which killed more than 1,300 people, mostly New Orleanians. Inland regional state and local healthcare and human services agencies rushed to assist evacuees, most of whom were uninsured or displaced without employer healthcare coverage. The initial evacuation brought more than 350,000 evacuees seeking shelter to the greater Baton Rouge area, LA, 80 miles north of New Orleans, the closest high ground. This investigation describes the rapid needs assessment developed and conducted by the Capital Area Human Services District of the greater Baton Rouge area, a quasi-governmental human services authority, the regional provider of state-funded mental health, addictive disorders, and developmental disabilities services, on a sample of 6,553 Katrina evacuees in the greater Baton Rouge area. In the event of catastrophic natural and manmade disasters, state and federal decision makers should follow the National Incident Management System and support local designated lead agencies with additional resources as requested. They must rely on designated lead agencies to use their knowledge of the locale, local resources, and relationships with other providers and volunteers to respond rapidly and efficiently to evacuee needs identified through a designated, concise tool that is singularly utilized across the impacted region by all providers to determine the needed response.

  20. Growing Pains

    CERN Multimedia

    Katarina Anthony

    2013-01-01

    Heat expands and cold contracts: it’s a simple thermodynamic rule. But when temperatures swing from 300 K to near-absolute zero, this rule can mean a contraction of more than 80 metres across the LHC’s 27-km-long cryogenic system. Keeping this growth in check are compensators (a.k.a. bellows), which shrink and stretch in response to thermodynamic changes. Leak tests and X-rays now underway in the tunnel have revealed that these “joints” might be suffering from growing pains…   This 25-μm weld crack is thought to be the cause of the helium leaks. Prior to the LS1 warm-up, CERN’s cryogenic experts knew of two points in the machine’s cryogenic distribution system that were leaking helium. Fortunately, these leaks were sufficiently small, confined to known sub-sectors of the cryogenic line and – with help from the vacuum team (TE-VSC) – could easily be compensated for. But as the machine warmed up f...

  1. Pluripotent stem cell-derived organoids: using principles of developmental biology to grow human tissues in a dish.

    Science.gov (United States)

    McCauley, Heather A; Wells, James M

    2017-03-15

    Pluripotent stem cell (PSC)-derived organoids are miniature, three-dimensional human tissues generated by the application of developmental biological principles to PSCs in vitro The approach to generate organoids uses a combination of directed differentiation, morphogenetic processes, and the intrinsically driven self-assembly of cells that mimics organogenesis in the developing embryo. The resulting organoids have remarkable cell type complexity, architecture and function similar to their in vivo counterparts. In the past five years, human PSC-derived organoids with components of all three germ layers have been generated, resulting in the establishment of a new human model system. Here, and in the accompanying poster, we provide an overview of how principles of developmental biology have been essential for generating human organoids in vitro , and how organoids are now being used as a primary research tool to investigate human developmental biology. © 2017. Published by The Company of Biologists Ltd.

  2. Three-Dimensional Evaluation of the Upper Airway Morphological Changes in Growing Patients with Skeletal Class III Malocclusion Treated by Protraction Headgear and Rapid Palatal Expansion: A Comparative Research.

    Directory of Open Access Journals (Sweden)

    Xueling Chen

    Full Text Available The aim of this study was to evaluate the morphological changes of upper airway after protraction headgear and rapid maxillary expansion (PE treatment in growing patients with Class III malocclusion and maxillary skeletal deficiency compared with untreated Class III patients by cone-beam computed tomography (CBCT.Thirty growing patients who have completed PE therapy were included in PE group. The control group (n = 30 was selected from the growing untreated patients with the same diagnosis. The CBCT scans of the pre-treatment (T1 and post-treatment (T2 of PE group and the control group were collected. Reconstruction and registration of the 3D models of T1 and T2 were completed. By comparing the data obtained from T1, T2 and control group, the morphological changes of the upper airway during the PE treatment were evaluated.Comparing with the data from T1 group, the subspinale (A of maxilla and the upper incisor (UI of the T2 group were moved in the anterior direction. The gnathion (Gn of mandible was moved in the posterior-inferior direction. The displacement of the hyoid bone as well as the length and width of dental arch showed significant difference. The volume and mean cross-sectional area of nasopharynx, velopharynx and glossopharynx region showed significant difference. The largest anteroposterior/the largest lateral (AP/LR ratios of the velopharynx and glossopharynx were increased, but the AP/LR ratio of the hypopharynx was decreased. In addition, the length and width of the maxillary dental arch, the displacement of the hyoid bone, the volume of nasopharynx and velopharynx, and the AP/LR ratio of the hypopharynx and velopharynx showed significant difference between the data from control and T2 group.The PE treatment of Class III malocclusion with maxillary skeletal hypoplasia leads to a significant increase in the volume of nasopharynx and velopharynx.

  3. Three-Dimensional Evaluation of the Upper Airway Morphological Changes in Growing Patients with Skeletal Class III Malocclusion Treated by Protraction Headgear and Rapid Palatal Expansion: A Comparative Research.

    Science.gov (United States)

    Chen, Xueling; Liu, Dongxu; Liu, Ju; Wu, Zizhong; Xie, Yongtao; Li, Liang; Liu, Hong; Guo, Tiantian; Chen, Chen; Zhang, Shijie

    2015-01-01

    The aim of this study was to evaluate the morphological changes of upper airway after protraction headgear and rapid maxillary expansion (PE) treatment in growing patients with Class III malocclusion and maxillary skeletal deficiency compared with untreated Class III patients by cone-beam computed tomography (CBCT). Thirty growing patients who have completed PE therapy were included in PE group. The control group (n = 30) was selected from the growing untreated patients with the same diagnosis. The CBCT scans of the pre-treatment (T1) and post-treatment (T2) of PE group and the control group were collected. Reconstruction and registration of the 3D models of T1 and T2 were completed. By comparing the data obtained from T1, T2 and control group, the morphological changes of the upper airway during the PE treatment were evaluated. Comparing with the data from T1 group, the subspinale (A) of maxilla and the upper incisor (UI) of the T2 group were moved in the anterior direction. The gnathion (Gn) of mandible was moved in the posterior-inferior direction. The displacement of the hyoid bone as well as the length and width of dental arch showed significant difference. The volume and mean cross-sectional area of nasopharynx, velopharynx and glossopharynx region showed significant difference. The largest anteroposterior/the largest lateral (AP/LR) ratios of the velopharynx and glossopharynx were increased, but the AP/LR ratio of the hypopharynx was decreased. In addition, the length and width of the maxillary dental arch, the displacement of the hyoid bone, the volume of nasopharynx and velopharynx, and the AP/LR ratio of the hypopharynx and velopharynx showed significant difference between the data from control and T2 group. The PE treatment of Class III malocclusion with maxillary skeletal hypoplasia leads to a significant increase in the volume of nasopharynx and velopharynx.

  4. Upper body contributions to power generation during rapid, overhand throwing in humans.

    Science.gov (United States)

    Roach, Neil T; Lieberman, Daniel E

    2014-06-15

    High-speed and accurate throwing is a distinctive human behavior. Achieving fast projectile speeds during throwing requires a combination of elastic energy storage at the shoulder, as well as the transfer of kinetic energy from proximal body segments to distal segments. However, the biomechanical bases of these mechanisms are not completely understood. We used inverse dynamics analyses of kinematic data from 20 baseball players fitted with four different braces that inhibit specific motions to test a model of power generation at key joints during the throwing motion. We found that most of the work produced during throwing is generated at the hips, and much of this work (combined with smaller contributions from the pectoralis major) is used to load elastic elements in the shoulder and power the rapid acceleration of the projectile. Despite rapid angular velocities at the elbow and wrist, the restrictions confirm that much of the power generated to produce these distal movements comes from larger proximal segments, such as the shoulder and torso. Wrist hyperextension enhances performance only modestly. Together, our data also suggest that heavy reliance on elastic energy storage may help explain some common throwing injuries and can provide further insight into the evolution of the upper body and when our ancestors first developed the ability to produce high-speed throws. © 2014. Published by The Company of Biologists Ltd.

  5. Human activity accelerating the rapid desertification of the Mu Us Sandy Lands, North China

    Science.gov (United States)

    Miao, Yunfa; Jin, Heling; Cui, Jianxin

    2016-01-01

    Over the past several thousand years, arid and semiarid China has experienced a series of asynchronous desertification events in its semiarid sandy and desert regions, but the precise identification of the driving forces of such events has remained elusive. In this paper we identify two rapid desertification events (RDEs) at ~4.6 ± 0.2 ka BP and ~3.3 ± 0.2 ka BP from the JJ Profile, located in the eastern Mu Us Sandy Lands. These RDEs appear to have occurred immediately following periods marked by persistently frequent and intense fires. We argue that such fire patterns, directly linked to an uncontrolled human use of vegetation as fuel, played a key role in accelerating RDEs by ensuring that the land surface was degraded beyond the threshold required for rapid desertification. This would suggest that the future use of a massive and sustained ecological program of vegetation rehabilitation should reduce the risk of destructive fire. PMID:26961705

  6. Direct comparison between confocal and multiphoton microscopy for rapid histopathological evaluation of unfixed human breast tissue

    Science.gov (United States)

    Yoshitake, Tadayuki; Giacomelli, Michael G.; Cahill, Lucas C.; Schmolze, Daniel B.; Vardeh, Hilde; Faulkner-Jones, Beverly E.; Connolly, James L.; Fujimoto, James G.

    2016-12-01

    Rapid histopathological examination of surgical specimen margins using fluorescence microscopy during breast conservation therapy has the potential to reduce the rate of positive margins on postoperative histopathology and the need for repeat surgeries. To assess the suitability of imaging modalities, we perform a direct comparison between confocal fluorescence microscopy and multiphoton microscopy for imaging unfixed tissue and compare to paraffin-embedded histology. An imaging protocol including dual channel detection of two contrast agents to implement virtual hematoxylin and eosin images is introduced that provides high quality imaging under both one and two photon excitation. Corresponding images of unfixed human breast tissue show that both confocal and multiphoton microscopy can reproduce the appearance of conventional histology without the need for physical sectioning. We further compare normal breast tissue and invasive cancer specimens imaged at multiple magnifications, and assess the effects of photobleaching for both modalities using the staining protocol. The results demonstrate that confocal fluorescence microscopy is a promising and cost-effective alternative to multiphoton microscopy for rapid histopathological evaluation of ex vivo breast tissue.

  7. Rapid and sensitive liquid chromatography-mass spectrometry method for determination of ropinirole in human plasma.

    Science.gov (United States)

    Bhatt, Jignesh; Jangid, Arvind; Shetty, Raghavendra; Shah, Bhavin; Kambli, Sandeep; Subbaiah, Gunta; Singh, Sadhana

    2006-03-18

    A rapid and robust liquid chromatography-mass spectrometry (LC-MS/MS) method was developed for non-ergoline dopamine D(2)-receptor agonist, ropinirole in human plasma using Es-citalopram oxalate as an internal standard. The method involves solid phase extraction from plasma, reversed-phase simple isocratic chromatographic conditions and mass spectrometric detection that enables a detection limit at picogram levels. The proposed method was validated with linear range of 20-1,200 pg/ml. The extraction recoveries for ropinirole and internal standard were 90.45 and 65.42%, respectively. The R.S.D.% of intra-day and inter-day assay was lower than 15%. For its sensitivity and reliability, the proposed method is particularly suitable for pharmacokinetic studies.

  8. Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy.

    Science.gov (United States)

    Li, Mi; Xiao, Xiubin; Liu, Lianqing; Xi, Ning; Wang, Yuechao

    2016-09-01

    Understanding the physicochemical properties of cell surface signalling molecules is important for us to uncover the underlying mechanisms that guide the cellular behaviors. Atomic force microscopy (AFM) has become a powerful tool for detecting the molecular interactions on individual cells with nanometer resolution. In this paper, AFM peak force tapping (PFT) imaging mode was applied to rapidly locate and visually map the CD20 molecules on human lymphoma cells using biochemically sensitive tips. First, avidin-biotin system was used to test the effectiveness of using PFT imaging mode to probe the specific molecular interactions. The adhesion images obtained on avidin-coated mica using biotin-tethered tips obviously showed the recognition spots which corresponded to the avidins in the simultaneously obtained topography images. The experiments confirmed the specificity and reproducibility of the recognition results. Then, the established procedure was applied to visualize the nanoscale organization of CD20s on the surface of human lymphoma Raji cells using rituximab (a monoclonal anti-CD20 antibody)-tethered tips. The experiments showed that the recognition spots in the adhesion images corresponded to the specific CD20-rituximab interactions. The cluster sizes of CD20s on lymphoma Raji cells were quantitatively analyzed from the recognition images. Finally, under the guidance of fluorescence recognition, the established procedure was applied to cancer cells from a clinical lymphoma patient. The results showed that there were significant differences between the adhesion images obtained on cancer cells and on normal cells (red blood cell). The CD20 distributions on ten cancer cells from the patient were quantified according to the adhesion images. The experimental results demonstrate the capability of applying PFT imaging to rapidly investigate the nanoscale biophysical properties of native membrane proteins on the cell surface, which is of potential significance in

  9. Rapid Turnover of Effector–Memory CD4+ T Cells in Healthy Humans

    Science.gov (United States)

    Macallan, Derek C.; Wallace, Diana; Zhang, Yan; de Lara, Catherine; Worth, Andrew T.; Ghattas, Hala; Griffin, George E.; Beverley, Peter C.L.; Tough, David F.

    2004-01-01

    Memory T cells can be divided into central–memory (TCM) and effector–memory (TEM) cells, which differ in their functional properties. Although both subpopulations can persist long term, it is not known whether they are maintained by similar mechanisms. We used in vivo labeling with deuterated glucose to measure the turnover of CD4+ T cells in healthy humans. The CD45R0+CCR7− TEM subpopulation was shown to have a rapid proliferation rate of 4.7% per day compared with 1.5% per day for CD45R0+CCR7+ TCM cells; these values are equivalent to average intermitotic (doubling) times of 15 and 48 d, respectively. In contrast, the CD45RA+CCR7+ naive CD4+ T cell population was found to be much longer lived, being labeled at a rate of only 0.2% per day (corresponding to an intermitotic time of approximately 1 yr). These data indicate that human CD4+ TEM cells constitute a short-lived cell population that requires continuous replenishment in vivo. PMID:15249595

  10. Rapid telomere motions in live human cells analyzed by highly time-resolved microscopy

    Directory of Open Access Journals (Sweden)

    Wang Xueying

    2008-10-01

    Full Text Available Abstract Background Telomeres cap chromosome ends and protect the genome. We studied individual telomeres in live human cancer cells. In capturing telomere motions using quantitative imaging to acquire complete high-resolution three-dimensional datasets every second for 200 seconds, telomere dynamics were systematically analyzed. Results The motility of individual telomeres within the same cancer cell nucleus was widely heterogeneous. One class of internal heterochromatic regions of chromosomes analyzed moved more uniformly and showed less motion and heterogeneity than telomeres. The single telomere analyses in cancer cells revealed that shorter telomeres showed more motion, and the more rapid telomere motions were energy dependent. Experimentally increasing bulk telomere length dampened telomere motion. In contrast, telomere uncapping, but not a DNA damaging agent, methyl methanesulfonate, significantly increased telomere motion. Conclusion New methods for seconds-scale, four-dimensional, live cell microscopic imaging and data analysis, allowing systematic tracking of individual telomeres in live cells, have defined a previously undescribed form of telomere behavior in human cells, in which the degree of telomere motion was dependent upon telomere length and functionality.

  11. Rapid flow cytometric measurement of cytokine-induced phosphorylation pathways [CIPP] in human peripheral blood leukocytes.

    Science.gov (United States)

    Montag, David T; Lotze, Michael T

    2006-11-01

    Current strategies designed to assess cells in the peripheral blood are limited to evaluation of phenotype or delayed measurement [>6 h] of function, usually quantifying cytokine production, cytolytic activity, or response to antigens. We reasoned that measurable abnormalities in signaling pathways could reflect pathological environs that cells experience in the setting of inflammatory states/cancer and could be represented in the peripheral blood. Two major pathways regulating the immune response are the JAK/STAT and MAPK/ERK pathways. These pathways are initiated by ligand-receptor binding and are rapidly propagated by subsequent protein phosphorylation cascades. We evaluated the brief application of cytokines in vitro to interrogate the early phosphorylation events of these signaling pathways in normal peripheral blood mononuclear cells (PBMC). Individual cytokine doses and time intervals of treatment were assessed to identify conditions useful in a clinical laboratory and as an initial goal to induce maximal phosphorylation. Surprisingly, all of the STAT proteins assessed and ERK1/2 are maximally phosphorylated within 15 min in human PBMC simply following addition of cytokines without preactivation of the cells. At 2 h, cells typically return to their basal phosphorylation states. For most of the cytokines tested, increased phosphorylation directly correlated with increased concentrations of the individual cytokines. These strategies will enable robust development of simple blood analyses to identify normal levels as well as impairments in STAT and MAPK/ERK signaling pathways associated with various human disease states including acute and chronic inflammatory conditions throughout clinical immunology.

  12. Flow induced dispersion analysis rapidly quantifies proteins in human plasma samples.

    Science.gov (United States)

    Poulsen, Nicklas N; Andersen, Nina Z; Østergaard, Jesper; Zhuang, Guisheng; Petersen, Nickolaj J; Jensen, Henrik

    2015-07-07

    Rapid and sensitive quantification of protein based biomarkers and drugs is a substantial challenge in diagnostics and biopharmaceutical drug development. Current technologies, such as ELISA, are characterized by being slow (hours), requiring relatively large amounts of sample and being subject to cumbersome and expensive assay development. In this work a new approach for quantification based on changes in diffusivity is presented. The apparent diffusivity of an indicator molecule interacting with the protein of interest is determined by Taylor Dispersion Analysis (TDA) in a hydrodynamic flow system. In the presence of the analyte the apparent diffusivity of the indicator changes due to complexation. This change in diffusivity is used to quantify the analyte. This approach, termed Flow Induced Dispersion Analysis (FIDA), is characterized by being fast (minutes), selective (quantification is possible in a blood plasma matrix), fully automated, and being subject to a simple assay development. FIDA is demonstrated for quantification of the protein Human Serum Albumin (HSA) in human plasma as well as for quantification of an antibody against HSA. The sensitivity of the FIDA assay depends on the indicator-analyte dissociation constant which in favourable cases is in the sub-nanomolar to picomolar range for antibody-antigen interactions.

  13. Hippocampal size predicts rapid learning of a cognitive map in humans.

    Science.gov (United States)

    Schinazi, Victor R; Nardi, Daniele; Newcombe, Nora S; Shipley, Thomas F; Epstein, Russell A

    2013-06-01

    The idea that humans use flexible map-like representations of their environment to guide spatial navigation has a long and controversial history. One reason for this enduring controversy might be that individuals vary considerably in their ability to form and utilize cognitive maps. Here we investigate the behavioral and neuroanatomical signatures of these individual differences. Participants learned an unfamiliar campus environment over a period of three weeks. In their first visit, they learned the position of different buildings along two routes in separate areas of the campus. During the following weeks, they learned these routes for a second and third time, along with two paths that connected both areas of the campus. Behavioral assessments after each learning session indicated that subjects formed a coherent representation of the spatial structure of the entire campus after learning a single connecting path. Volumetric analyses of structural MRI data and voxel-based morphometry (VBM) indicated that the size of the right posterior hippocampus predicted the ability to use this spatial knowledge to make inferences about the relative positions of different buildings on the campus. An inverse relationship between gray matter volume and performance was observed in the caudate. These results suggest that (i) humans can rapidly acquire cognitive maps of large-scale environments and (ii) individual differences in hippocampal anatomy may provide the neuroanatomical substrate for individual differences in the ability to learn and flexibly use these cognitive maps. © 2013 Wiley Periodicals, Inc. Copyright © 2013 Wiley Periodicals, Inc.

  14. Ultrasensitive cardiac troponin I antibody based nanohybrid sensor for rapid detection of human heart attack.

    Science.gov (United States)

    Bhatnagar, Deepika; Kaur, Inderpreet; Kumar, Ashok

    2017-02-01

    An ultrasensitive cardiac troponin I antibody conjugated with graphene quantum dots (GQD) and polyamidoamine (PAMAM) nanohybrid modified gold electrode based sensor was developed for the rapid detection of heart attack (myocardial infarction) in human. Screen printed gold (Au) electrode was decorated with 4-aminothiophenol for amine functionalization of the Au surface. These amino groups were further coupled with carboxyl functionalities of GQD with EDC-NHS reaction. In order to enhance the sensitivity of the sensor, PAMAM dendrimer was successively embedded on GQD through carbodiimide coupling to provide ultra-high surface area for antibody immobilization. The activated cardiac troponin I (cTnI) monoclonal antibody was immobilized on PAMAM to form nanoprobe for sensing specific heart attack marker cTnI. Various concentrations of cardiac marker, cTnI were electrochemically measured using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in human blood serum. The modifications on sensor surface were characterized by FTIR and AFM techniques. The sensor is highly specific to cTnI and showed negligible response to non-specific antigens. The sensitivity of the sensor was 109.23μAcm(-2)μg(-1) and lower limit of detection of cTnI was found 20fgmL(-1). Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Rapid and sensitive method for determination of withaferin-A in human plasma by HPLC.

    Science.gov (United States)

    Patial, Pankaj; Gota, Vikram

    2011-02-01

    To develop and validate a rapid and sensitive high-performance liquid chromatographic method for determination of withaferin-A in human plasma. Withaferin-A, the active molecule of a traditional Indian herb, has demonstrated several biological activities in preclinical models. A validated bioassay is not available for its pharmacokinetic evaluation. The chromatographic system used a reverse-phase C18 column with UV-visible detection at 225 nm. The mobile phase consisted of water and acetonitrile applied in a gradient flow. Withaferin-A was extracted by simple protein-precipitation technique. The calibration curve was linear in the concentration range of 0.05-1.6 µg/ml. The method has the desired sensitivity to detect the plasma concentration range of withaferin-A that is likely to show biological activity based on in vitro data. This is the first HPLC method ever described for the estimation of withaferin-A in human plasma which could be applied for pharmacokinetic studies.

  16. Learning new color names produces rapid increase in gray matter in the intact adult human cortex.

    Science.gov (United States)

    Kwok, Veronica; Niu, Zhendong; Kay, Paul; Zhou, Ke; Mo, Lei; Jin, Zhen; So, Kwok-Fai; Tan, Li Hai

    2011-04-19

    The human brain has been shown to exhibit changes in the volume and density of gray matter as a result of training over periods of several weeks or longer. We show that these changes can be induced much faster by using a training method that is claimed to simulate the rapid learning of word meanings by children. Using whole-brain magnetic resonance imaging (MRI) we show that learning newly defined and named subcategories of the universal categories green and blue in a period of 2 h increases the volume of gray matter in V2/3 of the left visual cortex, a region known to mediate color vision. This pattern of findings demonstrates that the anatomical structure of the adult human brain can change very quickly, specifically during the acquisition of new, named categories. Also, prior behavioral and neuroimaging research has shown that differences between languages in the boundaries of named color categories influence the categorical perception of color, as assessed by judgments of relative similarity, by response time in alternative forced-choice tasks, and by visual search. Moreover, further behavioral studies (visual search) and brain imaging studies have suggested strongly that the categorical effect of language on color processing is left-lateralized, i.e., mediated by activity in the left cerebral hemisphere in adults (hence "lateralized Whorfian" effects). The present results appear to provide a structural basis in the brain for the behavioral and neurophysiologically observed indices of these Whorfian effects on color processing.

  17. Characterization of human malignant mesothelioma cell lines orthotopically implanted in the pleural cavity of immunodeficient mice for their ability to grow and form metastasis

    Directory of Open Access Journals (Sweden)

    Orecchia Sara

    2006-05-01

    Full Text Available Abstract Background Malignant pleural mesothelioma (MPM is a tumor known to be resistant to conventional therapies. Thus, an in vivo model can represent an important tool for assessing the efficacy of novel approaches in the treatment of MPM. Presently, human MPM cells have been grown orthotopically in mice upon transplantation of tumor masses or tumor cell suspensions following surgery. In these models however, surgery can interfere with the tumor growth and the early stages of tumor development cannot be easily explored. Finally, results may not be so accurate due to implantation of potentially different tumor samples in different experimental groups. Our work aimed at establishing a nude mouse model xenotransplanted with human MPM cell lines in which tumor progression exhibits some features of the human disease. Methods Three distinct human MPM cell lines previously established from MPM patients displaying two different phenotypes, biphasic (MM-B1 and IST-Mes3 and epithelioid (IST-Mes2, were directly injected into the pleural cavity of nude mice. At different times, mice were sacrificed for autopsy, tumor nodules were counted and then removed for histology. Presence of metastases in visceral organs was also monitored. Results IST-Mes2 cells were unable to grow in nude mice. MM-B1 and IST-Mes3 cells were capable of growing in nude mice and formed tumor nodules in the pleura. Post-mortem examination showed that MPM cells progressively colonized the parietal and visceral pleura, the diaphragm, the mediastinum and, lastly the lung parenchyma. No pneumo-thorax was evidenced in the mice. Pleural effusions as well as lymph node metastases were observed only at later times. Conclusion This model mimics the progression of human malignant mesothelioma and it is easy to perform and reproducible; therefore it can be useful to study human MPM biology and evaluate the efficacy of novel therapies.

  18. How we learn to make decisions: rapid propagation of reinforcement learning prediction errors in humans.

    Science.gov (United States)

    Krigolson, Olav E; Hassall, Cameron D; Handy, Todd C

    2014-03-01

    Our ability to make decisions is predicated upon our knowledge of the outcomes of the actions available to us. Reinforcement learning theory posits that actions followed by a reward or punishment acquire value through the computation of prediction errors-discrepancies between the predicted and the actual reward. A multitude of neuroimaging studies have demonstrated that rewards and punishments evoke neural responses that appear to reflect reinforcement learning prediction errors [e.g., Krigolson, O. E., Pierce, L. J., Holroyd, C. B., & Tanaka, J. W. Learning to become an expert: Reinforcement learning and the acquisition of perceptual expertise. Journal of Cognitive Neuroscience, 21, 1833-1840, 2009; Bayer, H. M., & Glimcher, P. W. Midbrain dopamine neurons encode a quantitative reward prediction error signal. Neuron, 47, 129-141, 2005; O'Doherty, J. P. Reward representations and reward-related learning in the human brain: Insights from neuroimaging. Current Opinion in Neurobiology, 14, 769-776, 2004; Holroyd, C. B., & Coles, M. G. H. The neural basis of human error processing: Reinforcement learning, dopamine, and the error-related negativity. Psychological Review, 109, 679-709, 2002]. Here, we used the brain ERP technique to demonstrate that not only do rewards elicit a neural response akin to a prediction error but also that this signal rapidly diminished and propagated to the time of choice presentation with learning. Specifically, in a simple, learnable gambling task, we show that novel rewards elicited a feedback error-related negativity that rapidly decreased in amplitude with learning. Furthermore, we demonstrate the existence of a reward positivity at choice presentation, a previously unreported ERP component that has a similar timing and topography as the feedback error-related negativity that increased in amplitude with learning. The pattern of results we observed mirrored the output of a computational model that we implemented to compute reward

  19. Teaching Human Genetics with Mustard: Rapid Cycling "Brassica rapa" (Fast Plants Type) as a Model for Human Genetics in the Classroom Laboratory

    Science.gov (United States)

    Wendell, Douglas L.; Pickard, Dawn

    2007-01-01

    We have developed experiments and materials to model human genetics using rapid cycling "Brassica rapa", also known as Fast Plants. Because of their self-incompatibility for pollination and the genetic diversity within strains, "B. rapa" can serve as a relevant model for human genetics in teaching laboratory experiments. The experiment presented…

  20. Evaluation of oral fluid enzyme immunoassay for confirmation of a positive rapid human immunodeficiency virus test result.

    Science.gov (United States)

    Wesolowski, L G; Sanchez, T; MacKellar, D A; Branson, B M; Ethridge, S F; Constantine, N; Ketema, F; Sullivan, P S

    2009-07-01

    The CDC recommends that a reactive rapid human immunodeficiency virus (HIV) test be confirmed with an approved supplemental test; the performance of an intermediate enzyme immunoassay (EIA) is optional. In support of this recommendation, it was found that of 1,431 reactive rapid HIV test results, 2 (0.1%) had false-negative oral fluid Western blot results and both had false-negative EIA results.

  1. Evaluation of Oral Fluid Enzyme Immunoassay for Confirmation of a Positive Rapid Human Immunodeficiency Virus Test Result▿

    OpenAIRE

    Wesolowski, L. G.; Sanchez, T.; MacKellar, D. A.; Branson, B. M.; Ethridge, S. F.; Constantine, N.; Ketema, F.; Sullivan, P.S.

    2009-01-01

    The CDC recommends that a reactive rapid human immunodeficiency virus (HIV) test be confirmed with an approved supplemental test; the performance of an intermediate enzyme immunoassay (EIA) is optional. In support of this recommendation, it was found that of 1,431 reactive rapid HIV test results, 2 (0.1%) had false-negative oral fluid Western blot results and both had false-negative EIA results.

  2. Rapid Identification of Emerging Human-Pathogenic Sporothrix Species with Rolling Circle Amplification

    Science.gov (United States)

    Rodrigues, Anderson M.; Najafzadeh, Mohammad J.; de Hoog, G. Sybren; de Camargo, Zoilo P.

    2015-01-01

    Sporothrix infections are emerging as an important human and animal threat among otherwise healthy patients, especially in Brazil and China. Correct identification of sporotrichosis agents is beneficial for epidemiological surveillance, enabling implementation of adequate public-health policies and guiding antifungal therapy. In areas of limited resources where sporotrichosis is endemic, high-throughput detection methods that are specific and sensitive are preferred over phenotypic methods that usually result in misidentification of closely related Sporothrix species. We sought to establish rolling circle amplification (RCA) as a low-cost screening tool for species-specific identification of human-pathogenic Sporothrix. We developed six species-specific padlock probes targeting polymorphisms in the gene encoding calmodulin. BLAST-searches revealed candidate probes that were conserved intraspecifically; no significant homology with sequences from humans, mice, plants or microorganisms outside members of Sporothrix were found. The accuracy of our RCA-based assay was demonstrated through the specificity of probe-template binding to 25 S. brasiliensis, 58 S. schenckii, 5 S. globosa, 1 S. luriei, 4 S. mexicana, and 3 S. pallida samples. No cross reactivity between closely related species was evident in vitro, and padlock probes yielded 100% specificity and sensitivity down to 3 × 106 copies of the target sequence. RCA-based speciation matched identifications via phylogenetic analysis of the gene encoding calmodulin and the rDNA operon (kappa 1.0; 95% confidence interval 1.0-1.0), supporting its use as a reliable alternative to DNA sequencing. This method is a powerful tool for rapid identification and specific detection of medically relevant Sporothrix, and due to its robustness has potential for ecological studies. PMID:26696992

  3. Rapid effects of phytoestrogens on human colonic smooth muscle are mediated by oestrogen receptor beta.

    LENUS (Irish Health Repository)

    Hogan, A M

    2012-02-01

    Epidemiological studies have correlated consumption of dietary phytoestrogens with beneficial effects on colon, breast and prostate cancers. Genomic and non-genomic mechanisms are responsible for anti-carcinogenic effects but, until now, the effect on human colon was assumed to be passive and remote. No direct effect on human colonic smooth muscle has previously been described. Institutional research board approval was granted. Histologically normal colon was obtained from the proximal resection margin of colorectal carcinoma specimens. Circular smooth muscle strips were microdissected and suspended under 1g of tension in organ baths containing oxygenated Krebs solution at 37 degrees C. After an equilibration period, tissues were exposed to diarylpropionitrile (DPN) (ER beta agonist) and 1,3,5-tris(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT) (ER alpha agonist) or to the synthetic phytoestrogen compounds genistein (n=8), daidzein (n=8), fisetin (n=8) and quercetin (n=8) in the presence or absence of fulvestrant (oestrogen receptor antagonist). Mechanism of action was investigated by inhibition of downstream pathways. The cholinergic agonist carbachol was used to induce contractile activity. Tension was recorded isometrically. Phytoestrogens inhibit carbachol-induced colonic contractility. In keeping with a non-genomic, rapid onset direct action, the effect was within minutes, reversible and similar to previously described actions of 17 beta oestradiol. No effect was seen in the presence of fulvestrant indicating receptor modulation. While the DPN exerted inhibitory effects, PPT did not. The effect appears to be reliant on a p38\\/mitogen activated protein kinase mediated induction of nitric oxide production in colonic smooth muscle. The present data set provides the first description of a direct effect of genistein, daidzein, fisetin and quercetin on human colonic smooth muscle. The presence of ER in colonic smooth muscle has been functionally proven and the beta

  4. Surface-stress sensors for rapid and ultrasensitive detection of active free drugs in human serum

    Science.gov (United States)

    Ndieyira, Joseph W.; Kappeler, Natascha; Logan, Stephen; Cooper, Matthew A.; Abell, Chris; McKendry, Rachel A.; Aeppli, Gabriel

    2014-03-01

    There is a growing appreciation that mechanical signals can be as important as chemical and electrical signals in biology. To include such signals in a systems biology description for understanding pathobiology and developing therapies, quantitative experiments on how solution-phase and surface chemistry together produce biologically relevant mechanical signals are needed. Because of the appearance of drug-resistant hospital `superbugs', there is currently great interest in the destruction of bacteria by bound drug-target complexes that stress bacterial cell membranes. Here, we use nanomechanical cantilevers as surface-stress sensors, together with equilibrium theory, to describe quantitatively the mechanical response of a surface receptor to different antibiotics in the presence of competing ligands in solution. The antibiotics examined are the standard, Food and Drug Administration-approved drug of last resort, vancomycin, and the yet-to-be approved oritavancin, which shows promise for controlling vancomycin-resistant infections. The work reveals variations among strong and weak competing ligands, such as proteins in human serum, that determine dosages in drug therapies. The findings further enhance our understanding of the biophysical mode of action of the antibiotics and will help develop better treatments, including choice of drugs as well as dosages, against pathogens.

  5. Structurally functional changes in the microbiota of nutrient solution with addition of liquid human wastes, used for growing plants in a closed ecological system

    Science.gov (United States)

    Tirranen, L. S.; Borodina, E. V.; Markov, A. A.

    The investigations have proved the dependence of microbial community of nutrient solution development upon the specific conditions of a closed human life-support ecological system: the time of using permanent solution, introduction of additives into the nutrient medium and system gas exchange closure. For the first time, liquid human wastes were introduced into the nutrient solution to increase the mass exchange closure. Correlation analysis revealed the direct dependence between the time of liquid native human excretions introduction into the nutrient solution and the development of organisms participating in nitrogen transformation and growth of microflora potentially dangerous for humans and plants. With the help of correlation analysis it has been determined that particularly close connection exists between the duration of introduction of liquid human wastes and bacteria Escherichia coli, denitrificators, ammonificators and urobacteria. The correlation coefficient for these microbial groups was: r = 0,78. The investigations showed that by the end of experiment the microbial community of nutrient solution fulfilled the role of native urine destructor successfully. Thus, introduction of human native excretions (at 70% substitution of nitrate nitrogen with urine nitrogen) into the nutrient solution used for growing wheat monoculture in a closed ecosystem is possible.

  6. Rapid recent human evolution and the accumulation of balanced genetic polymorphisms.

    Science.gov (United States)

    Wills, Christopher

    2011-01-01

    All evolutionary change can be traced to alterations in allele frequencies in populations over time. DNA sequencing on a massive scale now permits us to follow the genetic consequences as our species has diverged from our close relatives and as we have colonized different parts of the world and adapted to them. But it has been difficult to disentangle natural selection from many other factors that alter frequencies. These factors include mutation and intragenic reciprocal recombination, gene conversion, segregation distortion, random drift, and gene flow between populations (these last two are greatly influenced by splits and coalescences of populations over time). The first part of this review examines recent studies that have had some success in dissecting out the role of natural selection, especially in humans and Drosophila. Among many examples, these studies include those that have followed the rapid evolution of traits that may permit adaptation to high altitude in Tibetan and Andean populations. In some cases, directional selection has been so strong that it may have swept alleles close to fixation in the span of a few thousand years, a rapidity of change that is also sometimes encountered in other organisms. The second part of the review summarizes data showing that remarkably few alleles have been carried completely to fixation during our recent evolution. Some of the alleles that have not reached fixation may be approaching new internal equilibria, which would indicate polymorphisms that are maintained by balancing selection. Finally, the review briefly examines why genetic polymorphisms, particularly those that are maintained by negative frequency dependence, are likely to have played an important role in the evolution of our species. A method is suggested for measuring the contribution of these polymorphisms to our gene pool. Such polymorphisms may add to the ability of our species to adapt to our increasingly complex and challenging environment.

  7. The Charlie Sheen Effect on Rapid In-home Human Immunodeficiency Virus Test Sales.

    Science.gov (United States)

    Allem, Jon-Patrick; Leas, Eric C; Caputi, Theodore L; Dredze, Mark; Althouse, Benjamin M; Noar, Seth M; Ayers, John W

    2017-07-01

    One in eight of the 1.2 million Americans living with human immunodeficiency virus (HIV) are unaware of their positive status, and untested individuals are responsible for most new infections. As a result, testing is the most cost-effective HIV prevention strategy and must be accelerated when opportunities are presented. Web searches for HIV spiked around actor Charlie Sheen's HIV-positive disclosure. However, it is unknown whether Sheen's disclosure impacted offline behaviors like HIV testing. The goal of this study was to determine if Sheen's HIV disclosure was a record-setting HIV prevention event and determine if Web searches presage increases in testing allowing for rapid detection and reaction in the future. Sales of OraQuick rapid in-home HIV test kits in the USA were monitored weekly from April 12, 2014, to April 16, 2016, alongside Web searches including the terms "test," "tests," or "testing" and "HIV" as accessed from Google Trends. Changes in OraQuick sales around Sheen's disclosure and prediction models using Web searches were assessed. OraQuick sales rose 95% (95% CI, 75-117; p < 0.001) of the week of Sheen's disclosure and remained elevated for 4 more weeks (p < 0.05). In total, there were 8225 more sales than expected around Sheen's disclosure, surpassing World AIDS Day by a factor of about 7. Moreover, Web searches mirrored OraQuick sales trends (r = 0.79), demonstrating their ability to presage increases in testing. The "Charlie Sheen effect" represents an important opportunity for a public health response, and in the future, Web searches can be used to detect and act on more opportunities to foster prevention behaviors.

  8. Reverse transcription genome exponential amplification reaction assay for rapid and universal detection of human rhinoviruses.

    Science.gov (United States)

    Guan, Li; Zhao, Lin-Qing; Zhou, Hang-Yu; Nie, Kai; Li, Xin-Na; Zhang, Dan; Song, Juan; Qian, Yuan; Ma, Xue-Jun

    2016-07-01

    Human rhinoviruses (HRVs) have long been recognized as the cause of more than one-half of acute viral upper respiratory illnesses, and they are associated with more-serious diseases in children, such as asthma, acute otitis media and pneumonia. A rapid and universal test for of HRV infection is in high demand. In this study, a reverse transcription genome exponential amplification reaction (RT-GEAR) assay targeting the HRV 5' untranslated region (UTR) was developed for pan-HRV detection. The reaction was performed in a single tube in one step at 65 °C for 60 min using a real-time fluorometer (Genie(®)II; Optigene). The RT-GEAR assay showed no cross-reactivity with common human enteroviruses, including HEV71, CVA16, CVA6, CVA10, CVA24, CVB5, Echo30, and PV1-3 or with other common respiratory viruses including FluA H3, FluB, PIV1-4, ADV3, RSVA, RSVB and HMPV. With in vitro-transcribed RNA containing the amplified regions of HRV-A60, HRV-B06 and HRV-C07 as templates, the sensitivity of the RT-GEAR assay was 5, 50 and 5 copies/reaction, respectively. Experiments to evaluate the clinical performance of the RT-GEAR assay were also carried out with a panel of 143 previously verified samples, and the results were compared with those obtained using a published semi-nested PCR assay followed by sequencing. The tested panel comprised 91 HRV-negative samples and 52 HRV-positive samples (18 HRV-A-positive samples, 3 HRV-B-positive samples and 31 HRV-C-positive samples). The sensitivity and specificity of the pan-HRVs RT-GEAR assay was 98.08 % and 100 %, respectively. The kappa correlation between the two methods was 0.985. The RT-GEAR assay based on a portable Genie(®)II fluorometer is a sensitive, specific and rapid assay for the universal detection of HRV infection.

  9. Neutrophil-mediated protection of cultured human vascular endothelial cells from damage by growing Candida albicans hyphae

    Energy Technology Data Exchange (ETDEWEB)

    Edwards, J.E. Jr.; Rotrosen, D.; Fontaine, J.W.; Haudenschild, C.C.; Diamond, R.D.

    1987-05-01

    Interactions were studied between human neutrophils and cultured human umbilical vein endothelial cells invaded by Candida albicans. In the absence of neutrophils, progressive Candida germination and hyphal growth extensively damaged endothelial cell monolayers over a period of 4 to 6 hours, as determined both by morphological changes and release of /sup 51/Cr from radiolabeled endothelial cells. Monolayers were completely destroyed and replaced by hyphae after 18 hours of incubation. In contrast, when added 2 hours after the monolayers had been infected with Candida, neutrophils selectively migrated toward and attached to hyphae at points of hyphal penetration into individual endothelial cells (observed by time-lapse video-microscopy). Attached neutrophils spread over hyphal surfaces both within and beneath the endothelial cells; neutrophil recruitment to initial sites of leukocyte-Candida-endothelial cell interactions continued throughout the first 60 minutes of observation. Neutrophil spreading and stasis were observed only along Candida hyphae and at sites of Candida-endothelial cell interactions. These events resulted in 58.0% killing of Candida at 2 hours and subsequent clearance of Candida from endothelial cell monolayers, as determined by microcolony counts and morphological observation. On introduction of additional neutrophils to yield higher ratios of neutrophils to endothelial cells (10 neutrophils:1 endothelial cell), neutrophil migration toward hyphal elements continued. Despite retraction or displacement of occasional endothelial cells by invading Candida and neutrophils, most endothelial cells remained intact, viable, and motile as verified both by morphological observations and measurement of /sup 51/Cr release from radiolabeled monolayers.

  10. Deterministically patterned biomimetic human iPSC-derived hepatic model via rapid 3D bioprinting

    Science.gov (United States)

    Ma, Xuanyi; Qu, Xin; Zhu, Wei; Li, Yi-Shuan; Yuan, Suli; Zhang, Hong; Liu, Justin; Wang, Pengrui; Lai, Cheuk Sun Edwin; Zanella, Fabian; Feng, Gen-Sheng; Sheikh, Farah; Chien, Shu; Chen, Shaochen

    2016-01-01

    The functional maturation and preservation of hepatic cells derived from human induced pluripotent stem cells (hiPSCs) are essential to personalized in vitro drug screening and disease study. Major liver functions are tightly linked to the 3D assembly of hepatocytes, with the supporting cell types from both endodermal and mesodermal origins in a hexagonal lobule unit. Although there are many reports on functional 2D cell differentiation, few studies have demonstrated the in vitro maturation of hiPSC-derived hepatic progenitor cells (hiPSC-HPCs) in a 3D environment that depicts the physiologically relevant cell combination and microarchitecture. The application of rapid, digital 3D bioprinting to tissue engineering has allowed 3D patterning of multiple cell types in a predefined biomimetic manner. Here we present a 3D hydrogel-based triculture model that embeds hiPSC-HPCs with human umbilical vein endothelial cells and adipose-derived stem cells in a microscale hexagonal architecture. In comparison with 2D monolayer culture and a 3D HPC-only model, our 3D triculture model shows both phenotypic and functional enhancements in the hiPSC-HPCs over weeks of in vitro culture. Specifically, we find improved morphological organization, higher liver-specific gene expression levels, increased metabolic product secretion, and enhanced cytochrome P450 induction. The application of bioprinting technology in tissue engineering enables the development of a 3D biomimetic liver model that recapitulates the native liver module architecture and could be used for various applications such as early drug screening and disease modeling. PMID:26858399

  11. Apnea-induced rapid eye movement sleep disruption impairs human spatial navigational memory.

    Science.gov (United States)

    Varga, Andrew W; Kishi, Akifumi; Mantua, Janna; Lim, Jason; Koushyk, Viachaslau; Leibert, David P; Osorio, Ricardo S; Rapoport, David M; Ayappa, Indu

    2014-10-29

    Hippocampal electrophysiology and behavioral evidence support a role for sleep in spatial navigational memory, but the role of particular sleep stages is less clear. Although rodent models suggest the importance of rapid eye movement (REM) sleep in spatial navigational memory, a similar role for REM sleep has never been examined in humans. We recruited subjects with severe obstructive sleep apnea (OSA) who were well treated and adherent with continuous positive airway pressure (CPAP). Restricting CPAP withdrawal to REM through real-time monitoring of the polysomnogram provides a novel way of addressing the role of REM sleep in spatial navigational memory with a physiologically relevant stimulus. Individuals spent two different nights in the laboratory, during which subjects performed timed trials before and after sleep on one of two unique 3D spatial mazes. One night of sleep was normally consolidated with use of therapeutic CPAP throughout, whereas on the other night, CPAP was reduced only in REM sleep, allowing REM OSA to recur. REM disruption via this method caused REM sleep reduction and significantly fragmented any remaining REM sleep without affecting total sleep time, sleep efficiency, or slow-wave sleep. We observed improvements in maze performance after a night of normal sleep that were significantly attenuated after a night of REM disruption without changes in psychomotor vigilance. Furthermore, the improvement in maze completion time significantly positively correlated with the mean REM run duration across both sleep conditions. In conclusion, we demonstrate a novel role for REM sleep in human memory formation and highlight a significant cognitive consequence of OSA. Copyright © 2014 the authors 0270-6474/14/3414571-07$15.00/0.

  12. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Directory of Open Access Journals (Sweden)

    Valentina Arfilli

    2015-11-01

    Full Text Available Shiga toxins (Stx have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h, radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins.

  13. Rapid 3D human ribcage and kidney modeling for transcostal HIFU surgery

    Science.gov (United States)

    Cao, Rui; Gao, Jing; Melzer, Andreas; Nabi, Ghulam; Huang, Zhihong

    2012-11-01

    A rapid modeling technique for constructing human ribcage and kidney models for high intensity focused ultrasound through the ribcage is proposed and tested. In this study, a 3D model reconstructed from a patient's CT images provides solutions for the conversion of 3D image data into multi-part volumetric models of kidney and ribcage. The model was imported into PZFlex for simulation of acoustic field analysis. Tissue mimicking materials for the phantom were selected based on the acoustic characterisation result. The effect of the ribcage on HIFU sonication and acoustic pressure distribution were measured in the focal plane. Porcine ribcage sample was used in experiments for comparison. Results showed that with the presence of the ribcage phantom, the maximum temperature at the focus was reduced by approximately 60-70% and the maximum pressure at the focal zone was halved. Focus splitting with the ribcage in place was demonstrated both in experiments and simulations. The development of this model provides basis for using patient's data for transcostal HIFU research and maximise the efficiency of the HIFU surgery.

  14. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

    Science.gov (United States)

    Arfilli, Valentina; Carnicelli, Domenica; Ardissino, Gianluigi; Torresani, Erminio; Scavia, Gaia; Brigotti, Maurizio

    2015-01-01

    Shiga toxins (Stx) have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC) strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h), radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins). PMID:26556372

  15. Rapid disease progression in human immunodeficiency virus type 1-infected individuals with adverse reactions to trimethoprim-sulfamethoxazole prophylaxis

    NARCIS (Netherlands)

    Veenstra, J.; Veugelers, P. J.; Keet, I. P.; van der Ven, A. J. A. M.; Miedema, F.; Lange, J. M.; Coutinho, R. A.

    1997-01-01

    We studied the relation between the occurrence of adverse reactions to trimethoprim-sulfamethoxazole (TMP-SMZ) prophylaxis and the subsequent course of human immunodeficiency virus (HIV) infection in a cohort of homosexual men. Adverse reactions to TMP-SMZ were associated with a more rapid

  16. Immunochromatographic Brucella-specific immunoglobulin M and G lateral flow assays for rapid serodiagnosis of human brucellosis

    NARCIS (Netherlands)

    Smits, Henk L.; Abdoel, Theresia H.; Solera, Javier; Clavijo, Encarnacion; Diaz, Ramon

    2003-01-01

    To fulfill the need for a simple and rapid diagnostic test for human brucellosis, we used the immunochromatographic lateral flow assay format to develop two assays, one for the detection of Brucella-specific immunoglobulin M (IgM) antibodies and one for the detection of Brucella-specific IgG

  17. Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization

    NARCIS (Netherlands)

    Mazza, G. (Giuseppe); Al-Akkad, W. (Walid); Telese, A. (Andrea); Longato, L. (Lisa); Urbani, L. (Luca); Robinson, B. (Benjamin); Hall, A. (Andrew); Kong, K. (Kenny); Frenguelli, L. (Luca); Marrone, G. (Giusi); Willacy, O. (Oliver); Shaeri, M. (Mohsen); A.J. Burns (Alan); Malago, M. (Massimo); Gilbertson, J. (Janet); Rendell, N. (Nigel); Moore, K. (Kevin); Hughes, D. (David); Notingher, I. (Ioan); Jell, G. (Gavin); Del Rio Hernandez, A. (Armando); P. de Coppi (Paolo); Rombouts, K. (Krista); Pinzani, M. (Massimo)

    2017-01-01

    textabstractThe development of human liver scaffolds retaining their 3-dimensional structure and extra-cellular matrix (ECM) composition is essential for the advancement of liver tissue engineering. We report the design and validation of a new methodology for the rapid and accurate production of

  18. Rapid identification of human SNAP-25 transcript variants by a miniaturized capillary electrophoresis system.

    Science.gov (United States)

    Németh, Nóra; Kerékgyártó, Márta; Sasvári-Székely, Mária; Rónai, Zsolt; Guttman, András

    2014-02-01

    The 25 kDa synaptosomal-associated protein (SNAP-25) is a crucial component of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex and plays an important role in neurotransmission in the central nervous system. SNAP-25 has two different splice variants, SNAP-25a and SNAP-25b, differing in nine amino acids that results in a slight functional alteration of the generated soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex. Two independent techniques, a PCR-miniaturized CE method and a real-time PCR based approach were elaborated for the specific and quantitative detection of the two SNAP-25 transcription variants. DNA-constructs coding for the two isoforms were used for optimization. Excellent specificity was observed with the use of our previously described highly sensitive miniaturized CE system in combination with quantitative PCR. The ratio of the two isoforms were reliably detected in a range of at least four orders of magnitude with a linear regression of R(2) = 0.987. Expression of the two isoforms was determined in human samples, where SNAP-25 was detected even in non-neural tissues, although at approximately a 100-fold lower level compared to the central nervous system. The relative amount of the SNAP-25b isoform was higher in the brain, whereas expression of SNAP-25a variant proved to be slightly higher in extra-neural cell types. The genomics approach in conjunction with the miniaturized CE system introduced in this paper is readily applicable for rapid alternative splice variant analysis. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Rapid Ganciclovir Susceptibility Assay Using Flow Cytometry for Human Cytomegalovirus Clinical Isolates

    Science.gov (United States)

    McSharry, James J.; Lurain, Nell S.; Drusano, George L.; Landay, Alan L.; Notka, Mostafa; O’Gorman, Maurice R. G.; Weinberg, Adriana; Shapiro, Howard M.; Reichelderfer, Patricia S.; Crumpacker, Clyde S.

    1998-01-01

    Rapid, quantitative, and objective determination of the susceptibilities of human cytomegalovirus (HCMV) clinical isolates to ganciclovir has been assessed by an assay that uses a fluorochrome-labeled monoclonal antibody to an HCMV immediate-early antigen and flow cytometry. Analysis of the ganciclovir susceptibilities of 25 phenotypically characterized clinical isolates by flow cytometry demonstrated that the 50% inhibitory concentrations (IC50s) of ganciclovir for 19 of the isolates were between 1.14 and 6.66 μM, with a mean of 4.32 μM (±1.93) (sensitive; IC50 less than 7 μM), the IC50s for 2 isolates were 8.48 and 9.79 μM (partially resistant), and the IC50s for 4 isolates were greater than 96 μM (resistant). Comparative analysis of the drug susceptibilities of these clinical isolates by the plaque reduction assay gave IC50s of less than 6 μM, with a mean of 2.88 μM (±1.40) for the 19 drug-sensitive isolates, IC50s of 6 to 8 μM for the partially resistant isolates, and IC50s of greater than 12 μM for the four resistant clinical isolates. Comparison of the IC50s for the drug-susceptible and partially resistant clinical isolates obtained by the flow cytometry assay with the IC50s obtained by the plaque reduction assay showed an acceptable correlation (r2 = 0.473; P = 0.001), suggesting that the flow cytometry assay could substitute for the more labor-intensive, subjective, and time-consuming plaque reduction assay. PMID:9736557

  20. Extracellular ATP induces the rapid release of HIV-1 from virus containing compartments of human macrophages.

    Science.gov (United States)

    Graziano, Francesca; Desdouits, Marion; Garzetti, Livia; Podini, Paola; Alfano, Massimo; Rubartelli, Anna; Furlan, Roberto; Benaroch, Philippe; Poli, Guido

    2015-06-23

    HIV type 1 (HIV-1) infects CD4(+) T lymphocytes and tissue macrophages. Infected macrophages differ from T cells in terms of decreased to absent cytopathicity and for active accumulation of new progeny HIV-1 virions in virus-containing compartments (VCC). For these reasons, infected macrophages are believed to act as "Trojan horses" carrying infectious particles to be released on cell necrosis or functional stimulation. Here we explored the hypothesis that extracellular ATP (eATP) could represent a microenvironmental signal potentially affecting virion release from VCC of infected macrophages. Indeed, eATP triggered the rapid release of infectious HIV-1 from primary human monocyte-derived macrophages (MDM) acutely infected with the CCR5-dependent HIV-1 strain. A similar phenomenon was observed in chronically infected promonocytic U1 cells differentiated to macrophage-like cells (D-U1) by costimulation with phorbol esters and urokinase-type plasminogen activator. Worthy of note, eATP did not cause necrotic, apoptotic, or pyroptotic cell death, and its effect on HIV-1 release was suppressed by Imipramine (an antidepressant agent known to inhibit microvesicle formation by interfering with membrane-associated acid sphingomyelinase). Virion release was not triggered by oxidized ATP, whereas the effect of eATP was inhibited by a specific inhibitor of the P2X7 receptor (P2X7R). Thus, eATP triggered the discharge of virions actively accumulating in VCC of infected macrophages via interaction with the P2X7R in the absence of significant cytopathicity. These findings suggest that the microvesicle pathway and P2X7R could represent exploitable targets for interfering with the VCC-associated reservoir of infectious HIV-1 virions in tissue macrophages.

  1. A rapid and quantitative method to detect human circulating tumor cells in a preclinical animal model.

    Science.gov (United States)

    Tu, Shih-Hsin; Hsieh, Yi-Chen; Huang, Li-Chi; Lin, Chun-Yu; Hsu, Kai-Wen; Hsieh, Wen-Shyang; Chi, Wei-Ming; Lee, Chia-Hwa

    2017-06-23

    As cancer metastasis is the deadliest aspect of cancer, causing 90% of human deaths, evaluating the molecular mechanisms underlying this process is the major interest to those in the drug development field. Both therapeutic target identification and proof-of-concept experimentation in anti-cancer drug development require appropriate animal models, such as xenograft tumor transplantation in transgenic and knockout mice. In the progression of cancer metastasis, circulating tumor cells (CTCs) are the most critical factor in determining the prognosis of cancer patients. Several studies have demonstrated that measuring CTC-specific markers in a clinical setting (e.g., flow cytometry) can provide a current status of cancer development in patients. However, this useful technique has rarely been applied in the real-time monitoring of CTCs in preclinical animal models. In this study, we designed a rapid and reliable detection method by combining a bioluminescent in vivo imaging system (IVIS) and quantitative polymerase chain reaction (QPCR)-based analysis to measure CTCs in animal blood. Using the IVIS Spectrum CT System with 3D-imaging on orthotropic-developed breast-tumor-bearing mice. In this manuscript, we established a quick and reliable method for measuring CTCs in a preclinical animal mode. The key to this technique is the use of specific human and mouse GUS primers on DNA/RNA of mouse peripheral blood under an absolute qPCR system. First, the high sensitivity of cancer cell detection on IVIS was presented by measuring the luciferase carried MDA-MB-231 cells from 5 to 5x10(11) cell numbers with great correlation (R(2) = 0.999). Next, the MDA-MB-231 cell numbers injected by tail vein and their IVIS radiance signals were strongly corrected with qPCR-calculated copy numbers (R(2) > 0.99). Furthermore, by applying an orthotropic implantation animal model, we successfully distinguished xenograft tumor-bearing mice and control mice with a significant difference (p < 0

  2. Mycobacterium lutetiense sp. nov., Mycobacterium montmartrense sp. nov. and Mycobacterium arcueilense sp. nov., members of a novel group of non-pigmented rapidly growing mycobacteria recovered from a water distribution system.

    Science.gov (United States)

    Konjek, Julie; Souded, Sabiha; Guerardel, Yann; Trivelli, Xavier; Bernut, Audrey; Kremer, Laurent; Welte, Benedicte; Joyeux, Michel; Dubrou, Sylvie; Euzeby, Jean-Paul; Gaillard, Jean-Louis; Sapriel, Guillaume; Heym, Beate

    2016-09-01

    From our recent survey of non-pigmented rapidly growing mycobacteria in the Parisian water system, three groups of isolates (taxons 1-3) corresponding to possible novel species were selected for taxonomic study. The three taxa each formed creamy white, rough colonies, had an optimal growth temperature of 30 °C, hydrolyzed Tween 80, were catalase-positive at 22 °C and expressed arylsulfatase activity. All three were susceptible to amikacin, ciprofloxacin and tigecycline. The three taxa produced specific sets of mycolic acids, including one family that has never previously been described, as determined by thin layer chromatography and nuclear magnetic resonance. The partial rpoB sequences (723 bp) showed 4-6 % divergence from each other and more than 5 % differences from the most similar species. Partial 16S rRNA gene sequences showed 99 % identity within each species. The most similar sequences for 16S rRNA genes (98-99 % identity over 1444-1461 bp) were found in the Mycobacterium fortuitum group, Mycobacterium septicum and Mycobacterium farcinogenes. The three taxa formed a new clade (bootstrap value, 99 %) on trees reconstructed from concatenated partial 16S rRNA, hsp65 and rpoB sequences. The above results led us to propose three novel species for the three groups of isolates, namely Mycobacterium lutetiense sp. nov. [type strain 071T=ParisRGMnew_1T (CIP 110656T=DSM 46713T)], Mycobacterium montmartrense sp. nov. [type strain 196T=ParisRGMnew_2T (CIP 110655T=DSM 46714T)] and Mycobacteriu marcueilense sp. nov. [type strain of 269T=ParisRGMnew_3T (CIP 110654T=DSM 46715T)].

  3. A simple and rapid Hepatitis A Virus (HAV titration assay based on antibiotic resistance of infected cells: evaluation of the HAV neutralization potency of human immune globulin preparations

    Directory of Open Access Journals (Sweden)

    Kaplan Gerardo G

    2008-12-01

    Full Text Available Abstract Background Hepatitis A virus (HAV, the causative agent of acute hepatitis in humans, is an atypical Picornaviridae that grows poorly in cell culture. HAV titrations are laborious and time-consuming because the virus in general does not cause cytopathic effect and is detected by immunochemical or molecular probes. Simple HAV titration assays could be developed using currently available viral construct containing selectable markers. Results We developed an antibiotic resistance titration assay (ARTA based on the infection of human hepatoma cells with a wild type HAV construct containing a blasticidin (Bsd resistance gene. Human hepatoma cells infected with the HAV-Bsd construct survived selection with 2 μg/ml of blasticidin whereas uninfected cells died within a few days. At 8 days postinfection, the color of the pH indicator phenol red in cell culture media correlated with the presence of HAV-Bsd-infected blasticidin-resistant cells: an orange-to-yellow color indicated the presence of growing cells whereas a pink-to-purple color indicated that the cells were dead. HAV-Bsd titers were determined by an endpoint dilution assay based on the color of the cell culture medium scoring orange-to-yellow wells as positive and pink-to-purple wells as negative for HAV. As a proof-of-concept, we used the ARTA to evaluate the HAV neutralization potency of two commercially available human immune globulin (IG preparations and a WHO International Standard for anti-HAV. The three IG preparations contained comparable levels of anti-HAV antibodies that neutralized approximately 1.5 log of HAV-Bsd. Similar neutralization results were obtained in the absence of blasticidin by an endpoint dilution ELISA at 2 weeks postinfection. Conclusion The ARTA is a simple and rapid method to determine HAV titers without using HAV-specific probes. We determined the HAV neutralization potency of human IG preparations in 8 days by ARTA compared to the 14 days required by the

  4. Multicenter evaluation of a new rapid automated human immunodeficiency virus antigen detection assay.

    Science.gov (United States)

    Weber, B; Mühlbacher, A; Michl, U; Paggi, G; Bossi, V; Sargento, C; Camacho, R; Fall, E H; Berger, A; Schmitt, U; Melchior, W

    1999-03-01

    Although human immunodeficiency virus (HIV) antigen assays are of limited value for monitoring antiretroviral therapy, they play an important role for confirmatory testing of fourth generation HIV screening enzyme immunoassay (EIA) reactive samples. In a multicenter study, a new automated rapid p24 antigen assay, Elecsys HIV Ag (Roche Diagnostics Boehringer Mannheim GmbH, Penzberg, Germany), was compared to FDA licensed tests (Abbott HIV-1 Ag monoclonal and Coulter HIV-1 p24 antigen assay). In the evaluation 27 seroconversion panels were included, sera from the acute phase of infection, single and follow-up samples from HIV antibody positive patients, dilution series of HIV antigen positive standards, sera and cell culture supernatants infected with different HIV-1 subtypes (A-H, and O) HIV-2 and recombinant HIV-1 (gag/env) isolates. To challenge the specificity of the new assay, 2565 unselected blood donors, sera from pregnant women, dialysis and hospitalized patients and 407 potentially cross-reactive samples were investigated. Acute HIV infection was detected in three to eight seroconversion panels earlier with Elecsys HIV Ag than with the alternative assays. Higher numbers of serum samples from HIV infected patients tested positive by Elecsys HIV Ag than with the comparative assays. All HIV-1 subtypes and HIV-2 isolates were recognized with Elecsys HIV Ag. Abbott HIV-1 Ag monoclonal and Coulter HIV-1 p24 antigen assay showed a variable sensitivity for the different HIV-1 subtypes. The specificity of Elecsys HIV Ag and Coulter HIV-1 p24 antigen assay were 99.8 and 99.93%, respectively. All the eight sera that were false reactive by Elecsys HIV Ag were tested negative with the Elecsys HIV Ag Neutralization Test. In conclusion, Elecsys HIV Ag was more sensitive than the alternative assays and showed a high specificity in combination with the neutralization assay. The very short incubation time of 18 min and the fully automated procedure of Elecsys HIV Ag which

  5. Easy xeno-free and feeder-free method for isolating and growing limbal stromal and epithelial stem cells of the human cornea.

    Directory of Open Access Journals (Sweden)

    Djida Ghoubay-Benallaoua

    Full Text Available Epithelial and stromal stem cells are required to maintain corneal transparency. The aim of the study was to develop a new method to isolate and grow both corneal stromal (SSC and epithelial limbal (LSC stem cells from small human limbal biopsies under culture conditions in accordance with safety requirements mandatory for clinical use in humans. Superficial limbal explants were retrieved from human donor corneo-scleral rims. Human limbal cells were dissociated by digestion with collagenase A, either after epithelial scraping or with no scraping. Isolated cells were cultured with Essential 8 medium (E8, E8 supplemented with EGF (E8+ or Green's medium with 3T3 feeder-layers. Cells were characterized by immunostaining, RT-qPCR, colony forming efficiency, sphere formation, population doubling, second harmonic generation microscopy and differentiation potentials. LSC were obtained from unscraped explants in E8, E8+ and Green's media and were characterized by colony formation and expression of PAX6, ΔNP63α, Bmi1, ABCG2, SOX9, CK14, CK15 and vimentin, with a few cells positive for CK3. LSC underwent 28 population doublings still forming colonies. SSC were obtained from both scraped and unscraped explants in E8 and E8+ media and were characterized by sphere formation, expression of PAX6, SOX2, BMI1, NESTIN, ABCG2, KERATOCAN, VIMENTIN, SOX9, SOX10 and HNK1, production of collagen fibrils and differentiation into keratocytes, fibroblasts, myofibroblasts, neurons, adipocytes, chondrocytes and osteocytes. SSC underwent 48 population doublings still forming spheres, Thus, this new method allows both SSC and LSC to be isolated from small superficial limbal biopsies and to be primary cultured in feeder-free and xeno-free conditions, which will be useful for clinical purposes.

  6. Easy xeno-free and feeder-free method for isolating and growing limbal stromal and epithelial stem cells of the human cornea.

    Science.gov (United States)

    Ghoubay-Benallaoua, Djida; de Sousa, Céline; Martos, Raphaël; Latour, Gaël; Schanne-Klein, Marie-Claire; Dupin, Elisabeth; Borderie, Vincent

    2017-01-01

    Epithelial and stromal stem cells are required to maintain corneal transparency. The aim of the study was to develop a new method to isolate and grow both corneal stromal (SSC) and epithelial limbal (LSC) stem cells from small human limbal biopsies under culture conditions in accordance with safety requirements mandatory for clinical use in humans. Superficial limbal explants were retrieved from human donor corneo-scleral rims. Human limbal cells were dissociated by digestion with collagenase A, either after epithelial scraping or with no scraping. Isolated cells were cultured with Essential 8 medium (E8), E8 supplemented with EGF (E8+) or Green's medium with 3T3 feeder-layers. Cells were characterized by immunostaining, RT-qPCR, colony forming efficiency, sphere formation, population doubling, second harmonic generation microscopy and differentiation potentials. LSC were obtained from unscraped explants in E8, E8+ and Green's media and were characterized by colony formation and expression of PAX6, ΔNP63α, Bmi1, ABCG2, SOX9, CK14, CK15 and vimentin, with a few cells positive for CK3. LSC underwent 28 population doublings still forming colonies. SSC were obtained from both scraped and unscraped explants in E8 and E8+ media and were characterized by sphere formation, expression of PAX6, SOX2, BMI1, NESTIN, ABCG2, KERATOCAN, VIMENTIN, SOX9, SOX10 and HNK1, production of collagen fibrils and differentiation into keratocytes, fibroblasts, myofibroblasts, neurons, adipocytes, chondrocytes and osteocytes. SSC underwent 48 population doublings still forming spheres, Thus, this new method allows both SSC and LSC to be isolated from small superficial limbal biopsies and to be primary cultured in feeder-free and xeno-free conditions, which will be useful for clinical purposes.

  7. Growing and Growing: Promoting Functional Thinking with Geometric Growing Patterns

    Science.gov (United States)

    Markworth, Kimberly A.

    2010-01-01

    Design research methodology is used in this study to develop an empirically-substantiated instruction theory about students' development of functional thinking in the context of geometric growing patterns. The two research questions are: (1) How does students' functional thinking develop in the context of geometric growing patterns? (2) What are…

  8. An RNA gene expressed during cortical development evolved rapidly in humans

    DEFF Research Database (Denmark)

    Pollard, Katherine S; Salama, Sofie R; Lambert, Nelle

    2006-01-01

    of the human brain. We devised a ranking of regions in the human genome that show significant evolutionary acceleration. Here we report that the most dramatic of these 'human accelerated regions', HAR1, is part of a novel RNA gene (HAR1F) that is expressed specifically in Cajal-Retzius neurons...

  9. Kinase pathway dependence in primary human leukemias determined by rapid inhibitor screening

    NARCIS (Netherlands)

    J.W. Tyner (Jeffrey); W.F. Yang (Wayne); A. Bankhead III (Armand); G. Fan (Guang); L.B. Fletcher (Luke); J. Bryant (Jade); J.M. Glover (Jason); B.H. Chang (Bill); S.E. Spurgeon (Stephen); W.H. Fleming (William); T. Kovacsovics; J. Gotlib (Jason); S.T. Oh (Stephen); M.W.N. Deininger (Michael W.); C.M. Zwaan (Christian Michel); M.L. den Boer (Monique); M.M. van den Heuvel-Eibrink (Marry); T. O'Hare (Thomas); B.J. Druker (Brian); M.M. Loriaux (Marc)

    2013-01-01

    textabstractKinases are dysregulated in most cancers, but the frequency of specific kinase mutations is low, indicating a complex etiology in kinase dysregulation. Here, we report a strategy to rapidly identify functionally important kinase targets, irrespective of the etiology of kinase pathway

  10. Reagent deposition for rapid multiplex pathogen identification in human blood culture samples

    DEFF Research Database (Denmark)

    Mogensen, Klaus Bo; Machado, Ana Manuel; Dufva, Martin

    2014-01-01

    viewed in a dual-color microscope configuration. The test takes 20-30 min to perform. In order to lower the cost of the test, rapid automated reagent deposition is needed. Here, ultrasonic spray coating of polyvinyl alcohol/PNA-probes on microscope glass slides is presented. Different wetting regimes...

  11. Prospective evaluation of three rapid diagnostic tests for diagnosis of human leptospirosis.

    NARCIS (Netherlands)

    M.G.A. Goris (Marga); M.M.G. Leeflang (Mariska); M. Lodén (Martin); J.F.P. Wagenaar (Jiri); P.R. Klatser (Paul); R.A. Hartskeerl (Rudy); K.R. Boer (Kimberly)

    2013-01-01

    markdownabstractDiagnosis of leptospirosis by the microscopic agglutination test (MAT) or by culture is confined to specialized laboratories. Although ELISA techniques are more common, they still require laboratory facilities. Rapid Diagnostic Tests (RDTs) can be used for easy point-of-care

  12. Evaluation of the clinical utility of a rapid blood test for human leptospirosis

    NARCIS (Netherlands)

    Eapen, C. K.; Sugathan, Sheela; Kuriakose, Mariamma; Abdoel, Theresia; Smits, Henk L.

    2002-01-01

    A rapid assay device for the detection of Leptospira-specific immunoglobulin M (IgM) antibodies was applied on whole blood samples collected from a group of consecutive patients admitted with clinical suspicion of leptospirosis to a district hospital in Kerala, India. The hospital is located in an

  13. Preclinical and clinical performance of the Efoora test, a rapid test for detection of human immunodeficiency virus-specific antibodies.

    Science.gov (United States)

    Arens, Max Q; Mundy, Linda M; Amsterdam, Daniel; Barrett, J Tom; Bigg, Dan; Bruckner, David; Hanna, Bruce; Prince, Harry; Purington, Timothy; Hanna, Todd; Hewitt, Ross; Kalinka, Carolyn; Koppes, Thomas; Maxwell, Sarz; Moe, Ardis; Doymaz, Mehmet; Poulter, Melinda; Saber-Tehrani, Maryam; Simard, Lorenzo; Wilkins-Carmody, Donna; Vidaver, John; Berger, Cheryl; Davis, Alan H; Alzona, Mortimer T

    2005-05-01

    Barriers to effective diagnostic testing for human immunodeficiency virus type 1 (HIV-1) infection can be reduced with simple, reliable, and rapid detection methods. Our objective was to determine the accuracy, sensitivity, and specificity of a new rapid, lateral-flow immunochromatographic HIV-1 antibody detection device. Preclinical studies were performed using seroconversion, cross-reaction, and interference panels, archived clinical specimens, and fresh whole blood. In a multicenter, prospective clinical trial, a four-sample matrix of capillary (fingerstick) whole-blood specimens and venous whole blood, plasma, and serum was tested for HIV-1 antibodies with the Efoora HIV rapid test (Efoora Inc., Buffalo Grove, IL) and compared with an enzyme immunoassay (EIA) (Abbott Laboratories) licensed by the Food and Drug Administration. Western blot and nucleic acid test supplemental assays were employed to adjudicate discordant samples. Preclinical testing of seroconversion panels showed that antibodies were often detected earlier by the rapid test than by a reference EIA. No significant interference or cross-reactions were observed. Testing of 4,984 archived specimens yielded a sensitivity of 99.2% and a specificity of 99.7%. A prospective multicenter clinical study with 2,954 adult volunteers demonstrated sensitivity and specificity for the Efoora HIV rapid test of 99.8% (95% confidence interval [CI], 99.3 and 99.98%) and 99.0% (95% CI, 98.5 and 99.4%), respectively. Reactive rapid HIV-1 antibody detection was confirmed in 99.6% of those with a known HIV infection (n = 939), 5.2% of those in the high-risk group (n = 1,003), and 0.1% of those in the low-risk group (n = 1,012). For 21 (0.71%) patients, there was discordance between the results of the rapid test and the confirmatory EIA/Western blot tests. We conclude that the Efoora HIV rapid test is a simple, rapid assay for detection of HIV-1 antibodies, with high sensitivity and specificity compared to a standardized

  14. Rapid fabricating technique for multi-layered human hepatic cell sheets by forceful contraction of the fibroblast monolayer.

    Directory of Open Access Journals (Sweden)

    Yusuke Sakai

    Full Text Available Cell sheet engineering is attracting attention from investigators in various fields, from basic research scientists to clinicians focused on regenerative medicine. However, hepatocytes have a limited proliferation potential in vitro, and it generally takes a several days to form a sheet morphology and multi-layered sheets. We herein report our rapid and efficient technique for generating multi-layered human hepatic cell (HepaRG® cell sheets using pre-cultured fibroblast monolayers derived from human skin (TIG-118 cells as a feeder layer on a temperature-responsive culture dish. Multi-layered TIG-118/HepaRG cell sheets with a thick morphology were harvested on day 4 of culturing HepaRG cells by forceful contraction of the TIG-118 cells, and the resulting sheet could be easily handled. In addition, the human albumin and alpha 1-antitrypsin synthesis activities of TIG-118/HepaRG cells were approximately 1.2 and 1.3 times higher than those of HepaRG cells, respectively. Therefore, this technique is considered to be a promising modality for rapidly fabricating multi-layered human hepatocyte sheets from cells with limited proliferation potential, and the engineered cell sheet could be used for cell transplantation with highly specific functions.

  15. Rapid reversal of innate immune dysregulation in blood of patients and livers of humanized mice with HCV following DAA therapy

    Science.gov (United States)

    Burchill, Matthew A.; Roby, Justin A.; Crochet, Nanette; Wind-Rotolo, Megan; Stone, Amy E.; Edwards, Michael G.; Dran, Rachael J.; Kriss, Michael S.; Gale, Michael

    2017-01-01

    Chronic hepatitis C virus (HCV) infection results in sustained immune activation in both the periphery and hepatic tissue. HCV infection induces innate immune signaling that is responsible for recognition of dsRNA, leading to activation of transcription factors and production of Type I and III IFNs, as well as pro-inflammatory cytokines and chemokines. Continued activation of host-immune mediated inflammation is thought to contribute to pathologic changes that result in progressive hepatic fibrosis. The current standard treatment for chronic HCV infection is directly-acting antivirals (DAAs), which have provided the unique opportunity to determine whether successful, rapid treatment-induced eradication of viral RNA normalizes the dysregulated antiviral innate immune response in patients chronically infected with HCV. Results First, in patients receiving two different combinations of DAAs, we found that DAAs induced not only rapid viral clearance, but also a re-setting of antiviral immune responses in the peripheral blood. Specifically, we see a rapid decline in the expression of genes associated with chronic IFN stimulation (IFIT3, USP18, IFIT1) as well as a rapid decline in genes associated with inflammation (IL1β, CXCL10, CXCL11) in the peripheral blood that precedes the complete removal of virus from the blood. Interestingly, this rapid reversal of innate immune activation was not seen in patients who successfully clear chronic HCV infection using IFN-based therapy. Next, using a novel humanized mouse model (Fah-/-RAG2-/-IL2rgnull—FRG), we assessed the changes that occur in the hepatic tissue following DAA treatment. DAA-mediated rapid HCV clearance resulted in blunting of the expression of proinflammatory responses while functionally restoring the RIG-I/MAVS axis in the liver of humanized mice. Conclusions Collectively, our data demonstrate that the rapid viral clearance following treatment with DAAs results in the rebalancing of innate antiviral response in

  16. High false-positive rate of human immunodeficiency virus rapid serum screening in a predominantly hispanic prenatal population.

    Science.gov (United States)

    Zacharias, Nikolaos M; Athanassaki, Ioanna D; Sangi-Haghpeykar, Haleh; Gardner, Michael O

    2004-12-01

    To identify the characteristics of the gravidas delivering at our birthing center that place them at risk for false-positive human immunodeficiency virus (HIV) enzyme-linked immunosorbent assay (ELISA). The medical records of all rapid HIV-ELISA-positive gravidas that delivered at our hospital between January 2000 and October 2001 were retrieved, and information was gathered regarding maternal demographics. The results of the Western blot tests were also retrieved and correlated to the ELISA results, across varying maternal characteristics. chi(2), Student's t-test and multivariate analysis were performed, as appropriate, using the SAS software; statistical significance was denoted by ppositive rapid HIV-ELISA out of 9,781 deliveries. Of those, 26 were confirmed as HIV infected by Western blot (overall HIV prevalence: 0.27%, ELISA-positive predictive value: 37.7%). The subgroup prevalence of HIV and positive predictive value of ELISA were 1.53 and 75% among Caucasians; 2.43 and 82.6% among African-Americans; and 0.05 and 9.8% among Hispanics, respectively (p or =5 lifetime) sexual partners was elicited in the majority of HIV-infected patients. The positive predictive value of rapid HIV-ELISA during pregnancy varies widely, depending on maternal race/ethnicity and sexual behavior. The routine disclosure of rapid intrapartum HIV serum screening results prior to Western blot confirmation should be avoided in very low-risk populations.

  17. The use of immunochromatographic rapid test for soft tissue remains identification in order to distinguish between human and non-human origin.

    Science.gov (United States)

    Gascho, Dominic; Morf, Nadja V; Thali, Michael J; Schaerli, Sarah

    2017-05-01

    Clear identification of soft tissue remains as being of non-human origin may be visually difficult in some cases e.g. due to decomposition. Thus, an additional examination is required. The use of an immunochromatographic rapid tests (IRT) device can be an easy solution with the additional advantage to be used directly at the site of discovery. The use of these test devices for detecting human blood at crime scenes is a common method. However, the IRT is specific not only for blood but also for differentiation between human and non-human soft tissue remains. In the following this method is discussed and validated by means of two forensic cases and several samples of various animals. Copyright © 2017 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.

  18. Detection of Elevated Signaling Amino Acids in Human Diabetic Vitreous by Rapid Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Miao-Jen Lu

    2007-01-01

    Full Text Available Elevated glutamate is implicated in the pathology of PDR. The ability to rapidly assess the glutamate and amino acid content of vitreous provides a more complete picture of the chemical changes occurring at the diabetic retina and may lead to a better understanding of the pathology of PDR. Vitreous humor was collected following vitrectomies of patients with PDR and control conditions of macular hole or epiretinal membrane. A capillary electrophoresis method was developed to quantify glutamate and arginine. The analysis is relatively fast (<6 minutes and utilizes a poly(ethyleneoxide and sodium dodecylsulfate run buffer. Both amino acid levels show significant increases in PDR patients versus controls and are comparable to other reports. The levels of vitreal glutamate vary inversely with the degree of observed hemorrhage. The results demonstrate a rapid method for assessment of a number of amino acids to characterize the chemical changes at the diabetic retina to better understand tissue changes and potentially identify new treatments.

  19. Growing media [Chapter 5

    Science.gov (United States)

    Douglass F. Jacobs; Thomas D. Landis; Tara Luna

    2009-01-01

    Selecting the proper growing medium is one of the most important considerations in nursery plant production. A growing medium can be defined as a substance through which roots grow and extract water and nutrients. In native plant nurseries, a growing medium can consist of native soil but is more commonly an "artificial soil" composed of materials such as peat...

  20. Rapid Identification of Emerging Human-Pathogenic Sporothrix Species with Rolling Circle Amplification

    NARCIS (Netherlands)

    Rodrigues, Anderson M; Najafzadeh, Mohammad J; de Hoog, G Sybren; de Camargo, Zoilo P

    2015-01-01

    Sporothrix infections are emerging as an important human and animal threat among otherwise healthy patients, especially in Brazil and China. Correct identification of sporotrichosis agents is beneficial for epidemiological surveillance, enabling implementation of adequate public-health policies and

  1. Time Domain Technique for Rapid, Broadband Measurement of Human Absorption Cross Section in a Reverberation Chamber

    OpenAIRE

    Robinson, Martin P; Zhang, Xiaotian; Flintoft, Ian D.

    2017-01-01

    Absorption cross section (ACS) of an object is used in stochastic power balance models, while human ACS is closely related to microwave dosimetry parameters such as specific absorption rate (SAR) and thus characterises exposure as well as effect of human bodies on multipath propagation. ACS, averaged over all directions of incidence, can be obtained in the frequency domain from the S-parameters of two antennas in a stirred-mode reverberation chamber; however, our new time domain method is fas...

  2. Rapid single-step induction of functional neurons from human pluripotent stem cells.

    Science.gov (United States)

    Zhang, Yingsha; Pak, Changhui; Han, Yan; Ahlenius, Henrik; Zhang, Zhenjie; Chanda, Soham; Marro, Samuele; Patzke, Christopher; Acuna, Claudio; Covy, Jason; Xu, Wei; Yang, Nan; Danko, Tamas; Chen, Lu; Wernig, Marius; Südhof, Thomas C

    2013-06-05

    Available methods for differentiating human embryonic stem cells (ESCs) and induced pluripotent cells (iPSCs) into neurons are often cumbersome, slow, and variable. Alternatively, human fibroblasts can be directly converted into induced neuronal (iN) cells. However, with present techniques conversion is inefficient, synapse formation is limited, and only small amounts of neurons can be generated. Here, we show that human ESCs and iPSCs can be converted into functional iN cells with nearly 100% yield and purity in less than 2 weeks by forced expression of a single transcription factor. The resulting ES-iN or iPS-iN cells exhibit quantitatively reproducible properties independent of the cell line of origin, form mature pre- and postsynaptic specializations, and integrate into existing synaptic networks when transplanted into mouse brain. As illustrated by selected examples, our approach enables large-scale studies of human neurons for questions such as analyses of human diseases, examination of human-specific genes, and drug screening. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. Transcriptome Signatures Reveal Rapid Induction of Immune-Responsive Genes in Human Memory CD8(+) T Cells.

    Science.gov (United States)

    Yang, Cheng; Khanniche, Asma; DiSpirito, Joanna R; Ji, Ping; Wang, Shujun; Wang, Ying; Shen, Hao

    2016-05-31

    Memory T cells (TM) play a prominent role in protection and auto-immunity due to their ability to mount a more effective response than naïve T cells (TN). However, the molecular mechanisms underlying enhanced functionality of TM are not well defined, particularly in human TM. We examined the global gene expression profiles of human CD8(+) TN and TM before and after stimulation. There were 1,284, 1,373 and 1,629 differentially expressed genes between TN and TM at 0 hr, 4 hr and 24 hr after stimulation, respectively, with more genes expressed to higher levels in TM. Genes rapidly up-regulated in TN cells were largely involved in nitrogen, nucleoside and amino acid metabolisms. In contrast, those in CD8(+) TM were significantly enriched for immune-response-associated processes, including cytokine production, lymphocyte activation and chemotaxis. Multiple cytokines were rapidly up-regulated in TM cells, including effector cytokines known to be produced by CD8(+) T cells and important for their functions, as well as regulatory cytokines, both pro- and anti-inflammatory, that are not typically produced by CD8(+) T cells. These results provide new insights into molecular mechanisms that contribute to the enhanced functionality of human CD8(+) TM and their prominent role in protection and auto-immunity.

  4. Effectiveness of travel restrictions in the rapid containment of human influenza: a systematic review

    Science.gov (United States)

    Mateus, Ana LP; Otete, Harmony E; Beck, Charles R; Dolan, Gayle P; Nguyen-Van-Tam, Jonathan S

    2014-01-01

    Abstract Objective To assess the effectiveness of internal and international travel restrictions in the rapid containment of influenza. Methods We conducted a systematic review according to the requirements of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Health-care databases and grey literature were searched and screened for records published before May 2014. Data extraction and assessments of risk of bias were undertaken by two researchers independently. Results were synthesized in a narrative form. Findings The overall risk of bias in the 23 included studies was low to moderate. Internal travel restrictions and international border restrictions delayed the spread of influenza epidemics by one week and two months, respectively. International travel restrictions delayed the spread and peak of epidemics by periods varying between a few days and four months. Travel restrictions reduced the incidence of new cases by less than 3%. Impact was reduced when restrictions were implemented more than six weeks after the notification of epidemics or when the level of transmissibility was high. Travel restrictions would have minimal impact in urban centres with dense populations and travel networks. We found no evidence that travel restrictions would contain influenza within a defined geographical area. Conclusion Extensive travel restrictions may delay the dissemination of influenza but cannot prevent it. The evidence does not support travel restrictions as an isolated intervention for the rapid containment of influenza. Travel restrictions would make an extremely limited contribution to any policy for rapid containment of influenza at source during the first emergence of a pandemic virus. PMID:25552771

  5. Rapid and efficient CRISPR/Cas9 gene inactivation in human neurons during human pluripotent stem cell differentiation and direct reprogramming.

    Science.gov (United States)

    Rubio, Alicia; Luoni, Mirko; Giannelli, Serena G; Radice, Isabella; Iannielli, Angelo; Cancellieri, Cinzia; Di Berardino, Claudia; Regalia, Giulia; Lazzari, Giovanna; Menegon, Andrea; Taverna, Stefano; Broccoli, Vania

    2016-11-18

    The CRISPR/Cas9 system is a rapid and customizable tool for gene editing in mammalian cells. In particular, this approach has widely opened new opportunities for genetic studies in neurological disease. Human neurons can be differentiated in vitro from hPSC (human Pluripotent Stem Cells), hNPCs (human Neural Precursor Cells) or even directly reprogrammed from fibroblasts. Here, we described a new platform which enables, rapid and efficient CRISPR/Cas9-mediated genome targeting simultaneously with three different paradigms for in vitro generation of neurons. This system was employed to inactivate two genes associated with neurological disorder (TSC2 and KCNQ2) and achieved up to 85% efficiency of gene targeting in the differentiated cells. In particular, we devised a protocol that, combining the expression of the CRISPR components with neurogenic factors, generated functional human neurons highly enriched for the desired genome modification in only 5 weeks. This new approach is easy, fast and that does not require the generation of stable isogenic clones, practice that is time consuming and for some genes not feasible.

  6. The human genome and sport, including epigenetics and athleticogenomics: a brief look at a rapidly changing field.

    Science.gov (United States)

    Sharp, N C Craig

    2008-09-01

    Since Hugh Montgomery discovered the first of what are now nearly 200 "fitness genes", together with rapid advances in human gene therapy, there is now a real prospect of the use of genes, genetic elements, and/or cells that have the capacity to enhance athletic performance (to paraphrase the World Anti-Doping Agency's definition of gene doping). This overview covers the main areas of interface between genetics and sport, attempts to provide a context against which gene doping may be viewed, and suggests a futuristic legitimate use of genomic (and possibly epigenetic) information in sport.

  7. Human adaptation responses to a rapidly changing Arctic: A research context for building system resilience

    Science.gov (United States)

    Chapin, T.; Brinkman, T. J.

    2016-12-01

    Although human behavior accounts for more uncertainty in future trajectories in climate change than do biophysical processes, most climate-change research fails to include human actions in research design and implementation. This is well-illustrated in the Arctic. At the global scale, arctic processes strongly influence the strength of biophysical feedbacks between global human emissions and the rate of climate warming. However, most human actions in the arctic have little effect on these feedbacks, so research can contribute most effectively to reduction in arctic warming through improved understanding of the strength of arctic-global biophysical feedbacks, as in NASA's ABoVE program, and its effective communication to policy makers and the public. In contrast, at the local to regional scale within the arctic, human actions may influence the ecological and societal consequences of arctic warming, so research benefits from active stakeholder engagement in research design and implementation. Human communities and other stakeholders (government and NGOs) respond heterogeneously to socioeconomic and environmental change, so research that documents the range of historical and current adaptive responses to change provides insights on the resilience (flexibility of future options) of social-ecological processes in the arctic. Alaskan communities have attempted a range of adaptive responses to coastal erosion (e.g., seasonal migration, protection in place, relocation), wildfire (fire suppression to use of fire to manage wildlife habitat or landscape heterogeneity), declining sea ice (e.g., new hunting technology, sea ice observations and predictions), and changes in wildlife and fish availability (e.g., switch to harvest of alternative species, harvest times, or harvest locations). Research that draws on both traditional and western knowledge facilitates adaptation and predictions of the likely societal consequences of climate change in the Arctic. Effective inclusion of

  8. Isolation of a human lymphoblastoid line heterozygous at the thymidine kinase locus: possibility for a rapid human cell mutation assay

    Energy Technology Data Exchange (ETDEWEB)

    Skopek, T.R.; Liber, H.L.; Penman, B.W.; Thilly, W.G.

    1978-09-29

    A thymidine kinase heterozygote designated H2BT has been isolated from the human lymphoblast line HH4. Significant increase in the trifluorothymidine-resistant fraction was observed in the new cell line following treatment with the mutagens ICR-191 and butylmethansulfonate. Phenotypic expression was complete forty-eight hours after treatment.

  9. Interferon-alpha treatment rapidly clears Hepatitis e virus infection in humanized mice

    NARCIS (Netherlands)

    M.D.B. van de Garde (Martijn D.B.); S.D. Pas (Suzan); G.W. van Oord (Gertine); L. Gama (Lucio); Choi, Y. (Youkyung); R.A. de Man (Robert); P.A. Boonstra (André); T. Vanwolleghem (Thomas)

    2017-01-01

    textabstractAntiviral treatment options for chronic Hepatitis E Virus (HEV) infections are limited and immunological determinants of viral persistence remain largely unexplored. We studied the antiviral potency of pegylated interferon-α (pegIFNα) against HEV infections in humanized mice and modelled

  10. An Optimized Shotgun Strategy for the Rapid Generation of Comprehensive Human Proteomes

    DEFF Research Database (Denmark)

    Bekker-Jensen, Dorte B; Kelstrup, Christian D; Batth, Tanveer S

    2017-01-01

    This study investigates the challenge of comprehensively cataloging the complete human proteome from a single-cell type using mass spectrometry (MS)-based shotgun proteomics. We modify a classical two-dimensional high-resolution reversed-phase peptide fractionation scheme and optimize a protocol ...

  11. Detection and monitoring of human bocavirus 1 infection by a new rapid antigen test

    NARCIS (Netherlands)

    Bruning, A. H. L.; Susi, P.; Toivola, H.; Christensen, A.; Söderlund-Venermo, M.; Hedman, K.; Aatola, H.; Zvirbliene, A.; Koskinen, J. O.

    2016-01-01

    Clinically relevant diagnosis of human bocavirus 1 (HBoV1) is challenging, as the virus is frequently detected in asymptomatic patients, and cofindings with other respiratory viruses are common. The clinical value of current diagnostic methods, such as PCR, is therefore low, and alternative

  12. Expandable and Rapidly Differentiating Human Induced Neural Stem Cell Lines for Multiple Tissue Engineering Applications

    Directory of Open Access Journals (Sweden)

    Dana M. Cairns

    2016-09-01

    Full Text Available Limited availability of human neurons poses a significant barrier to progress in biological and preclinical studies of the human nervous system. Current stem cell-based approaches of neuron generation are still hindered by prolonged culture requirements, protocol complexity, and variability in neuronal differentiation. Here we establish stable human induced neural stem cell (hiNSC lines through the direct reprogramming of neonatal fibroblasts and adult adipose-derived stem cells. These hiNSCs can be passaged indefinitely and cryopreserved as colonies. Independently of media composition, hiNSCs robustly differentiate into TUJ1-positive neurons within 4 days, making them ideal for innervated co-cultures. In vivo, hiNSCs migrate, engraft, and contribute to both central and peripheral nervous systems. Lastly, we demonstrate utility of hiNSCs in a 3D human brain model. This method provides a valuable interdisciplinary tool that could be used to develop drug screening applications as well as patient-specific disease models related to disorders of innervation and the brain.

  13. Rapid and sensitive liquid chromatography-tandem mass spectrometry method for the quantitation of levodropropizine in human plasma.

    Science.gov (United States)

    Tang, Yunbiao; Zhao, Limei; Wang, Yingwu; Fawcett, J Paul; Gu, Jingkai

    2005-05-05

    A rapid and sensitive LC-MS-MS method for quantifying levodropropizine in human plasma after oral administration of a single-dose (60 mg/day) was developed and validated. The sample preparation used liquid-liquid extraction with a mixture of dichloromethane-diethyl ether (2:3, v/v) in a basic environment. The retention time of levodropropizne and zolmitriptan (used as internal standard) was 1.6 and 1.4 min, respectively. The assay was linear over the range 0.25-500 ng/mL with a LOQ of 0.25 ng/mL. The intra- and inter-day precision were levodropropizine concentration profile in human plasma was determined.

  14. The precedence of syntax in the rapid emergence of human language in evolution as defined by the integration hypothesis.

    Directory of Open Access Journals (Sweden)

    Vitor eNóbrega

    2015-03-01

    Full Text Available Our core hypothesis is that the emergence of human language arose very rapidly from the linking of two pre-adapted systems found elsewhere in the animal world—an expression system, found, for example, in birdsong, and a lexical system, suggestively found in non-human primate calls (Miyagawa et al., 2013, 2014. We challenge the view that language has undergone a series of gradual changes—or a single preliminary protolinguistic stage—before achieving its full character. We argue that a full-fledged combinatorial operation Merge triggered the integration of these two pre-adapted systems, giving rise to a fully developed language. This goes against the gradualist view that there existed a structureless, protolinguistic stage, in which a rudimentary proto-Merge operation generated internally flat words. It is argued that compounds in present-day language are a fossilized form of this prior stage, a point which we will question.

  15. The precedence of syntax in the rapid emergence of human language in evolution as defined by the integration hypothesis.

    Science.gov (United States)

    Nóbrega, Vitor A; Miyagawa, Shigeru

    2015-01-01

    Our core hypothesis is that the emergence of human language arose very rapidly from the linking of two pre-adapted systems found elsewhere in the animal world-an expression system, found, for example, in birdsong, and a lexical system, suggestively found in non-human primate calls (Miyagawa et al., 2013, 2014). We challenge the view that language has undergone a series of gradual changes-or a single preliminary protolinguistic stage-before achieving its full character. We argue that a full-fledged combinatorial operation Merge triggered the integration of these two pre-adapted systems, giving rise to a fully developed language. This goes against the gradualist view that there existed a structureless, protolinguistic stage, in which a rudimentary proto-Merge operation generated internally flat words. It is argued that compounds in present-day language are a fossilized form of this prior stage, a point which we will question.

  16. Translating human genetics into mouse: the impact of ultra-rapid in vivo genome editing.

    Science.gov (United States)

    Aida, Tomomi; Imahashi, Risa; Tanaka, Kohichi

    2014-01-01

    Gene-targeted mutant animals, such as knockout or knockin mice, have dramatically improved our understanding of the functions of genes in vivo and the genetic diversity that characterizes health and disease. However, the generation of targeted mice relies on gene targeting in embryonic stem (ES) cells, which is a time-consuming, laborious, and expensive process. The recent groundbreaking development of several genome editing technologies has enabled the targeted alteration of almost any sequence in any cell or organism. These technologies have now been applied to mouse zygotes (in vivo genome editing), thereby providing new avenues for simple, convenient, and ultra-rapid production of knockout or knockin mice without the need for ES cells. Here, we review recent achievements in the production of gene-targeted mice by in vivo genome editing. © 2013 The Authors Development, Growth & Differentiation © 2013 Japanese Society of Developmental Biologists.

  17. Improvement and Evaluation of Loop-Mediated Isothermal Amplification for Rapid Detection of Toxoplasma gondii Infection in Human Blood Samples.

    Directory of Open Access Journals (Sweden)

    Xi-Meng Sun

    Full Text Available Loop-mediated isothermal amplification (LAMP, an attractive DNA amplification method, was developed as a valuable tool for the rapid detection of Toxoplasma gondii. In this study, species-specific LAMP primers were designed by targeting the AF146527 sequence, which was a conserved sequence of 200- to 300-fold repetitive 529 bp fragment of T.gondii. LAMP reaction system was optimized so that it could detect the minimal DNA sample such as a single tachyzoite or 10 copies of recombinant plasmid. No cross-reactivity was found when using DNA from other parasites as templates. Subsequently, a total of 200 human blood samples were directly investigated by two diagnostic methods, LAMP and conventional PCR. Fourteen of 200 (7% samples were positive for Toxoplasma by LAMP (the primers developed in this study, whereas only 5 of 200 (2.5% were proved positive by conventional PCR. The procedure of the LAMP assay was very simple, as the reaction would be carried out in a single tube under isothermal conditions at 64°C and the result would be read out with 1 h (as early as 35 min with loop primers. Thus, this method has the advantages of rapid amplification, simple operation, and easy detection and would be useful for rapid and reliable clinical diagnosis of acute toxoplasmosis, especially in developing countries.

  18. Foodborne transmission of bovine spongiform encephalopathy to non-human primates results in preclinical rapid-onset obesity.

    Directory of Open Access Journals (Sweden)

    Alexander Strom

    Full Text Available Obesity has become one of the largest public health challenges worldwide. Recently, certain bacterial and viral pathogens have been implicated in the pathogenesis of obesity. In the present study, we retrospectively analyzed clinical data, plasma samples and post-mortem tissue specimens derived from a risk assessment study in bovine spongiform encephalopathy (BSE-infected female cynomolgus monkeys (Macaca fascicularis. The original study design aimed to determine minimal infectious doses after oral or intracerebral (i.c. infection of macaques to assess the risk for humans. High-dose exposures resulted in 100% attack rates and a median incubation time of 4.7 years as described previously. Retrospective analyses of clinical data from high-dosed macaques revealed that foodborne BSE transmission caused rapid weight gain within 1.5 years post infection (β = 0.915; P<0.0001 which was not seen in age- and sex-matched control animals or i.c. infected animals. The rapid-onset obesity was not associated with impaired pancreatic islet function or glucose metabolism. In the early preclinical phase of oral transmission associated with body weight gain, prion accumulation was confined to the gastrointestinal tract. Intriguingly, immunohistochemical findings suggest that foodborne BSE transmission has a pathophysiological impact on gut endocrine cells which may explain rapid weight gain. To our knowledge, this is the first experimental model which clearly demonstrates that foodborne pathogens can induce obesity.

  19. The effects in humans of rapid loss of body mass on a boxing-related task.

    Science.gov (United States)

    Smith, M S; Dyson, R; Hale, T; Harrison, J H; McManus, P

    2000-09-01

    The physiological effects of strategies for a rapid loss of body mass immediately before weighing-in for competition in weight-governed sports are unclear. This study examined the effects of a 3%-4% loss in body mass on a boxing-related task. Seven novice amateur boxers completed three 3 min rounds of simulated boxing on a prototype boxing ergometer in an euhydrated state (E-trial) and after exercise-induced thermal dehydration (D-trial). All subjects lost body mass following dehydration-mean body mass fell 3.8 (SD +/- 0.3)% [77.3 (SD +/- 11.3) to 74.4 (SD +/- 10.7) kg, PPV) were inconsistent. Four subjects suffered reductions in PV between 15% and 30%, one subject maintained his E-trial value and two recorded an increase. The D-trial mean PV value was 8.0 (SD +/- 17.2)% lower but this fall was not statistically significant (P>0.05). Analysis of D-trial boxing performance showed one subject maintained his performance over the two trials and a second improved 17.8%. A two-way ANOVA (condition x time) with repeated measures on both factors showed no significant main effect differences for condition (F1,6 = 3.93 P>0.05), time (F1.83,48 = 1.12, P>0.05) or interaction between them (F1.93,48, P>0.05). Furthermore, neither heart rate nor blood lactate responses in the boxing task differed between trials. These data were affected by the small sample. Power and effect size analysis using eta(2) procedure and removing the outlier data produced a mean fall in boxing performance of 26.8%. However, some subjects appeared able to resist the deleterious effects of a rapid loss of body mass prior to competition and further research is needed to explain the mechanisms under-pinning this ability.

  20. Toward a mechanistic understanding of human-induced rapid environmental change: A case study linking energy development, avian nest predation, and predators

    Science.gov (United States)

    Hethcoat, Matthew G.; Chalfoun, Anna D.

    2015-01-01

    Demographic consequences of human-induced rapid environmental change (HIREC) have been widely documented for many populations. The mechanisms underlying such patterns, however, are rarely investigated and yet are critical to understand for effective conservation and management.

  1. BRaf signaling principles unveiled by large-scale human mutation analysis with a rapid lentivirus-based gene replacement method.

    Science.gov (United States)

    Lim, Chae-Seok; Kang, Xi; Mirabella, Vincent; Zhang, Huaye; Bu, Qian; Araki, Yoichi; Hoang, Elizabeth T; Wang, Shiqiang; Shen, Ying; Choi, Sukwoo; Kaang, Bong-Kiun; Chang, Qiang; Pang, Zhiping P; Huganir, Richard L; Zhu, J Julius

    2017-03-15

    Rapid advances in genetics are linking mutations on genes to diseases at an exponential rate, yet characterizing the gene mutation-cell behavior relationships essential for precision medicine remains a daunting task. More than 350 mutations on small GTPase BRaf are associated with various tumors, and ∼40 mutations are associated with the neurodevelopmental disorder cardio-facio-cutaneous syndrome (CFC). We developed a fast cost-effective lentivirus-based rapid gene replacement method to interrogate the physiopathology of BRaf and ∼50 disease-linked BRaf mutants, including all CFC-linked mutants. Analysis of simultaneous multiple patch-clamp recordings from 6068 pairs of rat neurons with validation in additional mouse and human neurons and multiple learning tests from 1486 rats identified BRaf as the key missing signaling effector in the common synaptic NMDA-R-CaMKII-SynGap-Ras-BRaf-MEK-ERK transduction cascade. Moreover, the analysis creates the original big data unveiling three general features of BRaf signaling. This study establishes the first efficient procedure that permits large-scale functional analysis of human disease-linked mutations essential for precision medicine. © 2017 Lim et al.; Published by Cold Spring Harbor Laboratory Press.

  2. Glycoblotting method allows for rapid and efficient glycome profiling of human Alzheimer's disease brain, serum and cerebrospinal fluid towards potential biomarker discovery.

    Science.gov (United States)

    Gizaw, Solomon T; Ohashi, Tetsu; Tanaka, Masakazu; Hinou, Hiroshi; Nishimura, Shin-Ichiro

    2016-08-01

    Understanding of the significance of posttranslational glycosylation in Alzheimer's disease (AD) is of growing importance for the investigation of the pathogenesis of AD as well as discovery research of the disease-specific serum biomarkers. We designed a standard protocol for the glycoblotting combined with MALDI-TOFMS to perform rapid and quantitative profiling of the glycan parts of glycoproteins (N-glycans) and glycosphingolipids (GSLs) using human AD's post-mortem samples such as brain tissues (dissected cerebral cortices such as frontal, parietal, occipital, and temporal domains), serum and cerebrospinal fluid (CSF). The structural profiles of the major N-glycans released from glycoproteins and the total expression levels of the glycans were found to be mostly similar between the brain tissues of the AD patients and those of the normal control group. In contrast, the expression levels of the serum and CSF protein N-glycans such as bisect-type and multiply branched glycoforms were increased significantly in AD patient group. In addition, the levels of some gangliosides such as GM1, GM2 and GM3 appeared to alter in the AD patient brain and serum samples when compared with the normal control groups. Alteration of the expression levels of major N- and GSL-glycans in human brain tissues, serum and CSF of AD patients can be monitored quantitatively by means of the glycoblotting-based standard protocols. The changes in the expression levels of the glycans derived from the human post-mortem samples uncovered by the standardized glycoblotting method provides potential serum biomarkers in central nervous system disorders and can contribute to the insight into the molecular mechanisms in the pathogenesis of neurodegenerative diseases and future drug discovery. Most importantly, the present preliminary trials using human post-mortem samples of AD patients suggest that large-scale serum glycomics cohort by means of various-types of human AD patients as well as the normal

  3. Rapid detection of structural variation in a human genome using nanochannel-based genome mapping technology

    DEFF Research Database (Denmark)

    Cao, Hongzhi; Hastie, Alex R.; Cao, Dandan

    2014-01-01

    BACKGROUND: Structural variants (SVs) are less common than single nucleotide polymorphisms and indels in the population, but collectively account for a significant fraction of genetic polymorphism and diseases. Base pair differences arising from SVs are on a much higher order (>100 fold) than poi...... mapping technology as a comprehensive and cost-effective method for detecting structural variation and studying complex regions in the human genome, as well as deciphering viral integration into the host genome.......BACKGROUND: Structural variants (SVs) are less common than single nucleotide polymorphisms and indels in the population, but collectively account for a significant fraction of genetic polymorphism and diseases. Base pair differences arising from SVs are on a much higher order (>100 fold) than point...... mutations; however, none of the current detection methods are comprehensive, and currently available methodologies are incapable of providing sufficient resolution and unambiguous information across complex regions in the human genome. To address these challenges, we applied a high-throughput, cost...

  4. Field Evaluation of the Determine Rapid Human Immunodeficiency Virus Diagnostic Test in Honduras and the Dominican Republic

    Science.gov (United States)

    Palmer, Carol J.; Dubon, Jose M.; Koenig, Ellen; Perez, Eddy; Ager, Arba; Jayaweera, Dushyantha; Cuadrado, Raul R.; Rivera, Ada; Rubido, Alex; Palmer, Dennis A.

    1999-01-01

    Rapid detection of human immunodeficiency virus (HIV) infection can result in improved patient care and/or faster implementation of public health preventive measures. A new rapid test, Determine (Abbott, Abbott Park, Ill.), detects HIV type 1 (HIV-1) and HIV-2 antibodies within 15 min by using 50 μl of serum or plasma. No specialized equipment or ancillary supplies are required, and results are read visually. A positive result is noted by the appearance of a red line. An operational control (red line) indicates proper test performance. We evaluated the Determine rapid HIV detection test with a group of well-characterized serum samples (CD4 counts and viral loads were known) and serum samples from HIV-positive individuals at field sites in Honduras and the Dominican Republic. In the field evaluations, the results obtained by the Determine assay were compared to those obtained by local in-country HIV screening procedures. We evaluated serum from 100 HIV-positive patients and 66 HIV-negative patients. All samples gave the expected results. In a companion study, 42 HIV-positive samples from a Miami, Fla., serum bank were tested by the Determine assay. The samples had been characterized in terms of CD4 counts and viral loads. Fifteen patients had CD4 counts 200 cells/mm3. Viral loads ranged from 630 to 873,746 log10 copies/ml. All samples from the Miami serum bank were positive by the Determine test. Combined results from the multicenter studies indicated that the correct results were obtained by the Determine assay for 100% (142 of 142) of the HIV-positive serum samples and 100% (66 of 66) of the HIV-negative serum samples. The Determine test was simple to perform and the results were easy to interpret. The Determine test provides a valuable new method for the rapid identification of HIV-positive individuals, especially in developing countries with limited laboratory infrastructures. PMID:10523577

  5. Use of Fourier transform infrared spectroscopy (FTIR spectroscopy for rapid and accurate identification of Yeasts isolated from human and animals

    Directory of Open Access Journals (Sweden)

    M. Taha

    2013-06-01

    Full Text Available Rapid and accurate identification of yeast is increasingly important to stipulate the appropriate therapy thus reducing morbidity and mortality related to yeast infections. Vibrational spectroscopic techniques (infrared (IR and Raman could provide potential alternatives to conventional typing methods, because they constitute a rapid, inexpensive and highly specific spectroscopic fingerprint through-which microorganism can be identified. The present study evaluate (FTIR spectroscopy as a sensitive and effective assay for the identification of the most frequent yeast species isolated from human and animals. One hundred and twenty-eight yeasts isolated from infected human mouths/vaginas, chronic diseased cows, crop mycosis in chicken and soil contaminated with pigeon droppings were phenotypically identified. Using universal primers, ITS1/ITS4, we have amplified ITS1-5.8S-ITS2 rDNA regions for 39 yeast isolates as representative samples. The PCR products were digested with restriction enzyme MspI and examined by PCR-RFLP, which was an efficient technique for identification of Candida spp., Cryptococcus neoformans and Trichosporon asahii. Further, identification of the same 39 isolates were done by FTIR spectroscopy and considered as reference for other strains by comparison of their FTIR spectra. The current study has sharply demonstrated the significant spectral differences between the various examined species of Candida, Cryptococcus, Trichosporon, Rhodotorula and Geotrichum isolated from different sources. Decisively, our research has confirmed that FTIR spectroscopy is a promising diagnostic tool, because of its sensitivity, rapidity, high differentiation capacity and simplicity compared to conventional/molecular techniques.

  6. Towards an ASEAN Human Rights Framework : Rapid Development in the Kingdom of Cambodia

    OpenAIRE

    Camilla Marie, Mårtensson

    2016-01-01

    The Association of Southeast Asia Nations (ASEAN) has since its origins in the late 1960’s been connected to the emphasis on Economic, Social and Cultural rights and the regional need for economic development. Through the intense years and debates that followed the first ASEAN Declaration the unity has made significant contributions to the Human Rights discourse due to the challenges and possibilities that surfaced. The Kingdom of Cambodia is one of the ASEAN member states poorest countries b...

  7. Rapid Detection of Trypanosoma cruzi in Human Serum by Use of an Immunochromatographic Dipstick Test ▿

    Science.gov (United States)

    Reithinger, Richard; Grijalva, Mario J.; Chiriboga, Rosa F.; Alarcón de Noya, Belkisyolé; Torres, Jaime R.; Pavia-Ruz, Norma; Manrique-Saide, Pablo; Cardinal, Marta V.; Gürtler, Ricardo E.

    2010-01-01

    We evaluated a commercially available immunochromatographic dipstick test to detect Trypanosoma cruzi infection in 366 human serum samples with known serological results from Argentina, Ecuador, Mexico, and Venezuela. One hundred forty-nine of 366 (40.7%) and 171/366 (46.7%) samples tested positive by dipstick and serology, respectively. Dipstick sensitivity was calculated to be 84.8% (range between countries, 77.5 to 95%), and specificity was 97.9% (95.9 to 100%). PMID:20534801

  8. Candida albicans and Candida parapsilosis rapidly up-regulate galectin-3 secretion by human gingival epithelial cells.

    Science.gov (United States)

    Tamai, Riyoko; Kiyoura, Yusuke

    2014-02-01

    Galectin-3 is a β-galactoside-binding C-type lectin that plays an important role in innate immunity. The purpose of this study was to determine whether Candida albicans and Candida parapsilosis up-regulate galectin-3 secretion by human gingival epithelial cells and gingival fibroblasts. Ca9-22, a human gingival epithelial cell line, and human gingival fibroblasts were incubated in the presence or absence of C. albicans or C. parapsilosis without serum. Levels of secreted human galectin-3 in culture supernatants were measured by enzyme-linked immunosorbent assay. We also pretreated Ca9-22 cells with cytochalasin D (an actin polymerization inhibitor), ALLN (a calpain inhibitor) and LY294002 [a phosphatidylinositol-3 kinase (PI3K) inhibitor] to determine whether the up-regulation of galectin-3 secretion was mediated by cytoskeletal changes, protease activity, or PI3K signaling. Galectin-3 secretion was significantly and rapidly up-regulated by live C. albicans and C. parapsilosis, as well as heat-killed C. albicans. In addition, cytochalasin D, LY294002 and ALLN did not inhibit the up-regulation in galectin-3 secretion. These results suggest that both live and heat-killed C. albicans and C. parapsilosis may increase the activity of the innate immune system and invasion by other microorganisms via up-regulation of galectin-3 secretion.

  9. A history into genetic and epigenetic evolution of food tolerance: how humanity rapidly evolved by drinking milk and eating wheat.

    Science.gov (United States)

    Blanchard, Carine

    2017-12-01

    Human exposure to wheat and milk is almost global worldwide. Yet the introduction of milk and wheat is very recent (5000-10 000 years) when compared to the human evolution. The last 4 decades have seen a rise in food allergy and food intolerance to milk and wheat. Often described as plurifactorial, the cause of allergic diseases is the result from an interplay between genetic predisposition and epigenetic in the context of environmental changes. Genetic and epigenetic understanding and their contribution to allergy or other antigen-driven diseases have considerably advanced in the last few years. Yet, environmental factors are also quite difficult to identify and associate with disease risk. Can we rethink our old findings and learn from human history and recent genetic studies? More than one million years separate Homo habilis to today's mankind, more than 1 million years to develop abilities to obtain food by foraging in diverse environments. One million year to adjust and fine-tune our genetic code and adapt; and only 1% of this time, 10 000 years, to face the three biggest revolutions of the human kind: the agricultural revolution, the industrial revolution and the postindustrial revolution. With big and rapid environmental changes come adaptation but with no time for fine-tuning. Today tolerance and adverse reactions to food may be a testimony of adaptation successes and mistakes.

  10. An Ambition to Grow

    OpenAIRE

    Ron Kemp; Hakkert, R.

    2006-01-01

    This report tries to gain insight in the willingness or ambition to grow of a small business owner. The main question of this report is therefore: Which factors influence the ambition to grow a business? To examine the ambition to grow an economic and a psychological perspective is given in this study.

  11. Rapid in vitro derivation of endothelium directly from human cancer cells.

    Directory of Open Access Journals (Sweden)

    Jennifer D Elster

    Full Text Available The development of an independent blood supply by a tumor is essential for maintaining growth beyond a certain limited size and for providing a portal for metastatic dissemination. Host-derived endothelial cells (ECs residing in and compromising the tumor vasculature originate via distinct processes known as sprouting angiogenesis and vasculogenesis. More recently ECs originating directly from the tumor cells themselves have been described although the basis for this phenomenon remains poorly understood. Here we describe in vitro conditions that allow lung and ovarian cancer cells to undergo a rapid and efficient transition into ECs that are indistinguishable from those obtained in vivo. A variety of methods were used to establish that the acquired phenotypes and behaviors of these tumor-derived ECs (TDECs closely resemble those of authentic ECs. Xenografts arising from co-inoculated in vitro-derived TDECs and tumor cells were also more highly vascularized than control tumors; moreover, their blood vessels were on average larger and frequently contained admixtures of host-derived ECs and TDECs derived from the initial inoculum. These results demonstrate that cancer cells can be manipulated under well-defined in vitro conditions to initiate a tumor cell-to-EC transition that is largely cell-autonomous, highly efficient and closely mimics the in vivo process. These studies provide a suitable means by which to identify and perhaps modify the earliest steps in TDEC generation.

  12. CS-SCORE: Rapid identification and removal of human genome contaminants from metagenomic datasets.

    Science.gov (United States)

    Haque, Mohammed Monzoorul; Bose, Tungadri; Dutta, Anirban; Reddy, Chennareddy Venkata Siva Kumar; Mande, Sharmila S

    2015-08-01

    Metagenomic sequencing data, obtained from host-associated microbial communities, are usually contaminated with host genome sequence fragments. Prior to performing any downstream analyses, it is necessary to identify and remove such contaminating sequence fragments. The time and memory requirements of available host-contamination detection techniques are enormous. Thus, processing of large metagenomic datasets is a challenging task. This study presents CS-SCORE--a novel algorithm that can rapidly identify host sequences contaminating metagenomic datasets. Validation results indicate that CS-SCORE is 2-6 times faster than the current state-of-the-art methods. Furthermore, the memory footprint of CS-SCORE is in the range of 2-2.5GB, which is significantly lower than other available tools. CS-SCORE achieves this efficiency by incorporating (1) a heuristic pre-filtering mechanism and (2) a directed-mapping approach that utilizes a novel sequence composition metric (cs-score). CS-SCORE is expected to be a handy 'pre-processing' utility for researchers analyzing metagenomic datasets. For academic users, an implementation of CS-SCORE is freely available at: http://metagenomics.atc.tcs.com/cs-score (or) https://metagenomics.atc.tcs.com/preprocessing/cs-score. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. An Optimized Shotgun Strategy for the Rapid Generation of Comprehensive Human Proteomes

    DEFF Research Database (Denmark)

    Bekker-Jensen, Dorte B; Kelstrup, Christian D; Batth, Tanveer S

    2017-01-01

    This study investigates the challenge of comprehensively cataloging the complete human proteome from a single-cell type using mass spectrometry (MS)-based shotgun proteomics. We modify a classical two-dimensional high-resolution reversed-phase peptide fractionation scheme and optimize a protocol......-coding genes). This depth is comparable with next-generation RNA sequencing and enables the identification of post-translational modifications, including ∼7,000 N-acetylation sites and ∼10,000 phosphorylation sites, without the need for enrichment. We further demonstrate the general applicability and clinical...

  14. The human small intestinal microbiota is driven by rapid uptake and conversion of simple carbohydrates

    DEFF Research Database (Denmark)

    Zoetendal, Erwin G; Raes, Jeroen; van den Bogert, Bartholomeus

    2012-01-01

    the microbiota at other parts of the GI tract, which is especially true for the small intestine because of its limited accessibility. Here we deduce an ecological model of the microbiota composition and function in the small intestine, using complementing culture-independent approaches. Phylogenetic microarray......The human gastrointestinal tract (GI tract) harbors a complex community of microbes. The microbiota composition varies between different locations in the GI tract, but most studies focus on the fecal microbiota, and that inhabiting the colonic mucosa. Consequently, little is known about...

  15. Rapid Assay for Simultaneous Detection and Differentiation of Immunoglobulin G Antibodies to Human Immunodeficiency Virus Type 1 (HIV-1) Group M, HIV-1 Group O, and HIV-2

    OpenAIRE

    Vallari, Ana S.; Hickman, Robert K.; Hackett, John R.; Brennan, Catherine A.; Varitek, Vincent A.; Devare, Sushil G.

    1998-01-01

    A rapid immunodiagnostic test that detects and discriminates human immunodeficiency virus (HIV) infections on the basis of viral type, HIV type 1 (HIV-1) group M, HIV-1 group O, or HIV-2, was developed. The rapid assay for the detection of HIV (HIV rapid assay) was designed as an instrument-free chromatographic immunoassay that detects immunoglobulin G (IgG) antibodies to HIV. To assess the performance of the HIV rapid assay, 470 HIV-positive plasma samples were tested by PCR and/or Western b...

  16. Rapid and Quantitative Assay of Amyloid-Seeding Activity in Human Brains Affected with Prion Diseases.

    Directory of Open Access Journals (Sweden)

    Hanae Takatsuki

    Full Text Available The infectious agents of the transmissible spongiform encephalopathies are composed of amyloidogenic prion protein, PrPSc. Real-time quaking-induced conversion can amplify very small amounts of PrPSc seeds in tissues/body fluids of patients or animals. Using this in vitro PrP-amyloid amplification assay, we quantitated the seeding activity of affected human brains. End-point assay using serially diluted brain homogenates of sporadic Creutzfeldt-Jakob disease patients demonstrated that 50% seeding dose (SD50 is reached approximately 10(10/g brain (values varies 10(8.79-10.63/g. A genetic case (GSS-P102L yielded a similar level of seeding activity in an autopsy brain sample. The range of PrPSc concentrations in the samples, determined by dot-blot assay, was 0.6-5.4 μg/g brain; therefore, we estimated that 1 SD50 unit was equivalent to 0.06-0.27 fg of PrPSc. The SD50 values of the affected brains dropped more than three orders of magnitude after autoclaving at 121°C. This new method for quantitation of human prion activity provides a new way to reduce the risk of iatrogenic prion transmission.

  17. Tachyphylaxis of human forearm vascular responses does not occur rapidly after exposure to isoproterenol.

    Science.gov (United States)

    Stein, C M; Nelson, R; Deegan, R; He, H; Inagami, T; Frazer, M; Badr, K F; Wood, M; Wood, A J

    1995-06-01

    In vitro and limited in vivo data suggest that rapid desensitization of beta-adrenoceptor responses occurs after exposure to agonist. Tachyphylaxis to a beta-adrenoceptor agonist would represent a potentially important mechanism for the short-term regulation of vascular tone. The effects of a 4-hour infusion of 400 ng/min intra-arterial isoproterenol on forearm blood flow and presynaptic beta-adrenoceptor-mediated norepinephrine release were determined in eight healthy volunteers. Intra-arterial isoproterenol at 400 ng/min resulted in a significant increase in forearm blood flow in all eight subjects at all time points, with no evidence of tachyphylaxis. In fact, forearm blood flow after 4 hours of the isoproterenol infusion (22.8 +/- 3.3 mL/100 mL per minute) was significantly greater than after 7 minutes (14.6 +/- 2.8 mL/100 mL per minute), 15 minutes (15.4 +/- 2.4 mL/100 mL per minute), and 30 minutes (17.4 +/- 3.0 mL/100 mL per minute) of the infusion (P < .05). Similarly, presynaptic beta-adrenoceptor responses showed no evidence of tachyphylaxis, so forearm norepinephrine spillover values after 7 minutes (6.6 +/- 0.94 ng/min), 15 minutes (7.6 +/- 1.5 ng/min), and 4 hours (8.8 +/- 1.1 ng/min) of isoproterenol infusion were increased and similar. Minimal systemic effects were observed, and there was no evidence of tolerance, there being no difference in heart rate after 7 minutes (70.7 +/- 2.7 beats per minute) and 4 hours (72.2 +/- 3.6 beats per minute) of isoproterenol infusion.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Prospective evaluation of three rapid diagnostic tests for diagnosis of human leptospirosis.

    Directory of Open Access Journals (Sweden)

    Marga G A Goris

    Full Text Available BACKGROUND: Diagnosis of leptospirosis by the microscopic agglutination test (MAT or by culture is confined to specialized laboratories. Although ELISA techniques are more common, they still require laboratory facilities. Rapid Diagnostic Tests (RDTs can be used for easy point-of-care diagnosis. This study aims to evaluate the diagnostic performance of the RDTs LeptoTek Dri Dot, LeptoTek Lateral Flow, and Leptocheck-WB, prospectively. METHODOLOGY: During 2001 to 2012, one or two of the RDTs at the same time have been applied prior to routine diagnostics (MAT, ELISA and culture on serum specimens from participants sent in for leptospirosis diagnosis. The case definition was based on MAT, ELISA and culture results. Participants not fulfilling the case definition were considered not to have leptospirosis. The diagnostic accuracy was determined based on the 1(st submitted sample and paired samples, either in an overall analysis or stratified according to days post onset of illness. RESULTS: The overall sensitivity and specificity for the LeptoTek Dri Dot was 75% respectively 96%, for the LeptoTek Lateral Flow 78% respectively 95%, and for the Leptocheck-WB 78% respectively 98%. Based on the 1(st submitted sample the sensitivity was low (51% for LeptoTek Dri Dot, 69% for LeptoTek Lateral Flow, and 55% for Leptocheck-WB, but substantially increased when the results of paired samples were combined, although accompanied by a lower specificity (82% respectively 91% for LeptoTek Dri Dot, 86% respectively 84% for LeptoTek Lateral Flow, and 80% respectively 93% for Leptocheck-WB. CONCLUSIONS: All three tests present antibody tests contributing to the diagnosis of leptospirosis, thus supporting clinical suspicion and contributing to awareness. Since the overall sensitivity of the tested RDTs did not exceed 80%, one should be cautious to rely only on an RDT result, and confirmation by reference tests is strongly recommended.

  19. Rapid analysis of clenbuterol, salbutamol, procaterol, and fenoterol in pharmaceuticals and human urine by capillary electrophoresis.

    Science.gov (United States)

    Sirichai, Somsak; Khanatharana, Proespichaya

    2008-09-15

    Capillary electrophoresis (CE) with UV detection for the simultaneous and short-time analysis of clenbuterol, salbutamol, procaterol, fenoterol is described and validated. Optimized conditions were found to be a 10 mmoll(-1) borate buffer (pH 10.0), an separation voltage of 19 kV, and a separation temperature of 32 degrees C. Detection was set at 205 nm. Under the optimized conditions, analyses of the four analytes in pharmaceutical and human urine samples were carried out in approximately 1 min. The interference of the sample matrix was not observed. The LOD (limits of detection) defined at S/N of 3:1 was found between 0.5 and 2.0 mgl(-1) for the analytes. The linearity of the detector response was within the range from 2.0 to 30 mgl(-1) with correlation coefficient >0.996.

  20. Application of a human monoclonal antibody in a rapid competitive anti-HIV ELISA.

    Science.gov (United States)

    Döpel, S H; Porstmann, T; Grunow, R; Jungbauer, A; Von Baehr, R

    1989-01-17

    The ELISA is the established screening technique for the detection of antibodies directed against HIV. The first generation assays, mostly based on the sandwich principle, employed purified virus from cell culture and gave both false-positive and false-negative results. Sandwich-type assays preferentially detect IgG antibodies, require a high serum dilution and are two-step procedures. In order to detect an immune response as early as possible after infection anti-HIV antibodies of the IgM class should also be measured. To this end a competitive ELISA has been developed using a solid phase-adsorbed recombinant HIV envelope protein and an enzyme-labelled human monoclonal antibody. This detects both IgM and IgG antibodies, the results are available within 1 h and a serum predilution is not necessary.

  1. Rapid Synthesis of Gold Nanoparticles from Quercus incana and Their Antimicrobial Potential against Human Pathogens

    Directory of Open Access Journals (Sweden)

    Rizwana Sarwar

    2017-01-01

    Full Text Available In current study, bioreduction of tetrachloroauric acid (HAuCl4·3H2O was carried out using leaves extract of Quercus incana for nanoparticle synthesis. The nanoparticles were characterized by ultraviolet visible spectrum (UV, Fourier-transform infrared (FT-IR, and transmission electron microscopy (TEM analysis. The gold nanoparticles (GNPs were generally clumpy agglomerates of polydispersed particles, with an average size in the range 5.5–10 nm. The Gas chromatography–mass spectrometry (GC–MS qualitative analysis and FT-IR data supported the presence of bioactive compounds, which are responsible for the metal reduction and nanoparticles stabilization. The biocompatibility of synthesized GNPs was evaluated via antibacterial activity by using human bacterial pathogens. The results showed that synthesized GNPs showed enhanced antibacterial activity against all bacterial pathogens.

  2. A rapid sound-action association effect in human insular cortex.

    Directory of Open Access Journals (Sweden)

    Isabella Mutschler

    Full Text Available BACKGROUND: Learning to play a musical piece is a prime example of complex sensorimotor learning in humans. Recent studies using electroencephalography (EEG and transcranial magnetic stimulation (TMS indicate that passive listening to melodies previously rehearsed by subjects on a musical instrument evokes differential brain activation as compared with unrehearsed melodies. These changes were already evident after 20-30 minutes of training. The exact brain regions involved in these differential brain responses have not yet been delineated. METHODOLOGY/PRINCIPAL FINDING: Using functional mri (fmri, we investigated subjects who passively listened to simple piano melodies from two conditions: in the 'actively learned melodies' condition subjects learned to play a piece on the piano during a short training session of a maximum of 30 minutes before the fMRI experiment, and in the 'passively learned melodies' condition subjects listened passively to and were thus familiarized with the piece. We found increased fMRI responses to actively compared with passively learned melodies in the left anterior insula, extending to the left fronto-opercular cortex. The area of significant activation overlapped the insular sensorimotor hand area as determined by our meta-analysis of previous functional imaging studies. CONCLUSIONS/SIGNIFICANCE: Our results provide evidence for differential brain responses to action-related sounds after short periods of learning in the human insular cortex. As the hand sensorimotor area of the insular cortex appears to be involved in these responses, re-activation of movement representations stored in the insular sensorimotor cortex may have contributed to the observed effect. The insular cortex may therefore play a role in the initial learning phase of action-perception associations.

  3. Human Adipose-Derived Stem Cells on Rapid Prototyped Three-Dimensional Hydroxyapatite/Beta-Tricalcium Phosphate Scaffold.

    Science.gov (United States)

    Canciani, Elena; Dellavia, Claudia; Ferreira, Lorena Maria; Giannasi, Chiara; Carmagnola, Daniela; Carrassi, Antonio; Brini, Anna Teresa

    2016-05-01

    In the study, we assess a rapid prototyped scaffold composed of 30/70 hydroxyapatite (HA) and beta-tricalcium-phosphate (β-TCP) loaded with human adipose-derived stem cells (hASCs) to determine cell proliferation, differentiation toward osteogenic lineage, adhesion and penetration on/into the scaffold.In this in vitro study, hASCs isolated from fat tissue discarded after plastic surgery were expanded, characterized, and then loaded onto the scaffold. Cells were tested for: viability assay (Alamar Blue at days 3, 7 and Live/Dead at day 32), differentiation index (alkaline phosphatase activity at day 14), scaffold adhesion (standard error of the mean analysis at days 5 and 18), and penetration (ground sections at day 32).All the hASC populations displayed stemness markers and the ability to differentiate toward adipogenic and osteogenic lineages.Cellular vitality increased between 3 and 7 days, and no inhibitory effect by HA/β-TCP was observed. Under osteogenic stimuli, scaffold increased alkaline phosphatase activity of +243% compared with undifferentiated samples. Human adipose-derived stem cells adhered on HA/β-TCP surface through citoplasmatic extensions that occupied the macropores and built networks among them. Human adipose derived stem cells were observed in the core of HA/β-TCP. The current combination of hASCs and HA/β-TCP scaffold provided encouraging results. If authors' data will be confirmed in preclinical models, the present engineering approach could represent an interesting tool in treating large bone defects.

  4. Rapid characterization of binding specificity and cross-reactivity of antibodies using recombinant human protein arrays.

    Science.gov (United States)

    Kijanka, Gregor; Ipcho, Simon; Baars, Sabine; Chen, Hong; Hadley, Katie; Beveridge, Allan; Gould, Edith; Murphy, Derek

    2009-01-30

    Antibodies are routinely used as research tools, in diagnostic assays and increasingly as therapeutics. Ideally, these applications require antibodies with high sensitivity and specificity; however, many commercially available antibodies are limited in their use as they cross-react with non-related proteins. Here we describe a novel method to characterize antibody specificity. Six commercially available monoclonal and polyclonal antibodies were screened on high-density protein arrays comprising of ~10,000 recombinant human proteins (Imagenes). Two of the six antibodies examined; anti-pICln and anti-GAPDH, bound exclusively to their target antigen and showed no cross-reactivity with non-related proteins. However, four of the antibodies, anti-HSP90, anti-HSA, anti-bFGF and anti-Ro52, showed strong cross-reactivity with other proteins on the array. Antibody-antigen interactions were readily confirmed using Western immunoblotting. In addition, the redundant nature of the protein array used, enabled us to define the epitopic region within HSP90 of the anti-HSP90 antibody, and identify possible shared epitopes in cross-reacting proteins. In conclusion, high-density protein array technology is a fast and effective means for determining the specificity of antibodies and can be used to further improve the accuracy of antibody applications.

  5. Evolution and behavioural responses to human-induced rapid environmental change

    Science.gov (United States)

    Sih, Andrew; Ferrari, Maud C O; Harris, David J

    2011-01-01

    Almost all organisms live in environments that have been altered, to some degree, by human activities. Because behaviour mediates interactions between an individual and its environment, the ability of organisms to behave appropriately under these new conditions is crucial for determining their immediate success or failure in these modified environments. While hundreds of species are suffering dramatically from these environmental changes, others, such as urbanized and pest species, are doing better than ever. Our goal is to provide insights into explaining such variation. We first summarize the responses of some species to novel situations, including novel risks and resources, habitat loss/fragmentation, pollutants and climate change. Using a sensory ecology approach, we present a mechanistic framework for predicting variation in behavioural responses to environmental change, drawing from models of decision-making processes and an understanding of the selective background against which they evolved. Where immediate behavioural responses are inadequate, learning or evolutionary adaptation may prove useful, although these mechanisms are also constrained by evolutionary history. Although predicting the responses of species to environmental change is difficult, we highlight the need for a better understanding of the role of evolutionary history in shaping individuals’ responses to their environment and provide suggestion for future work. PMID:25567979

  6. Control of HPNS in humans during rapid compression with trimix to 650 m (2131 ft).

    Science.gov (United States)

    Bennett, P B; Coggin, R; Roby, J

    1981-06-01

    In 1978 a series of deep trimix (He-N2-O2) dives was initiated to establish the relationship between a given nitrogen percentage and the rate of compression required to prevent the high pressure nervous syndrome (HPNS) at 460 m (1509 ft) and to determine the effects of inspired gas density, hydrostatic pressure, and narcosis o various circulatory and respiratory parameter, including the presence of dyspnea. In 1979, three human subjects were compressed to 460 m in 12 h 20 min with 5% N2 in He-O2. This resulted in nausea, vomiting, fatigue, tremors, and other signs and symptoms of HPNS that were especially prominent on arrival at that depth but had much improved by Day 2. In March 1980 the same profile was repeated but with 10% N2 in He-O2. The divers arrived at 460 m with virtually no symptoms of HPNS, but the psychometric performance, as for Atlantis I, still was decreased by some 40% on Day 1 and recovered to some 15% by Day 2. After 5-6 days at 460 m further extension of the dive to 650 m (2132 ft) with a 7.7% N2 mixture for 24 h showed similar control of symptoms of HPNS, although inspiratory resting dyspnea was present in one subject. The results are discussed in relation to the interactions of nitrogen percentage and rate of compression.

  7. A rapid lateral flow immunoassay for the detection of tyrosine phosphatase-like protein IA-2 autoantibodies in human serum.

    Directory of Open Access Journals (Sweden)

    Ingrid Kikkas

    Full Text Available Type 1 diabetes (T1D results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is subsequently captured on the anti-HA-Tag antibody-coated test line on the strip. The other site of the IA-2As is bound to biotinylated IA-2, allowing the complex to be visualized using colloidal gold nanoparticle-conjugated streptavidin. For this study, 35 serum samples from T1D patients and 44 control sera from non-diabetic individuals were analyzed with our novel assay and the results were correlated with two IA-2A ELISAs. Among the 35 serum samples from T1D patients, the IA-2A LFIA, the in-house IA-2A ELISA and the commercial IA-2A ELISA identified as positive 21, 29 and 30 IA-2A-positive sera, respectively. The major advantages of the IA-2A LFIA are its rapidity and simplicity.

  8. A rapid lateral flow immunoassay for the detection of tyrosine phosphatase-like protein IA-2 autoantibodies in human serum.

    Science.gov (United States)

    Kikkas, Ingrid; Mallone, Roberto; Larger, Etienne; Volland, Hervé; Morel, Nathalie

    2014-01-01

    Type 1 diabetes (T1D) results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As) are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA) based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is subsequently captured on the anti-HA-Tag antibody-coated test line on the strip. The other site of the IA-2As is bound to biotinylated IA-2, allowing the complex to be visualized using colloidal gold nanoparticle-conjugated streptavidin. For this study, 35 serum samples from T1D patients and 44 control sera from non-diabetic individuals were analyzed with our novel assay and the results were correlated with two IA-2A ELISAs. Among the 35 serum samples from T1D patients, the IA-2A LFIA, the in-house IA-2A ELISA and the commercial IA-2A ELISA identified as positive 21, 29 and 30 IA-2A-positive sera, respectively. The major advantages of the IA-2A LFIA are its rapidity and simplicity.

  9. A bioorganometallic approach for rapid electrochemical analysis of human immunodeficiency virus type-1 reverse transcriptase in serum

    Energy Technology Data Exchange (ETDEWEB)

    Labib, Mahmoud; Shipman, Patrick O.; Martic, Sanela [Department of Chemistry, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7 (Canada); Kraatz, Heinz-Bernhard, E-mail: hkraatz@uwo.ca [Department of Chemistry, University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7 (Canada)

    2011-05-30

    Highlights: > Two-dimensional electrochemical assay of HIV-1 reverse transcriptase in serum is presented. > XPS and ToF-SIMS confirmed binding of synthetic ferrocene-lipoic acid to gold. > Dynamic range was 100-500 pg mL{sup -1} with an LOD of 50 pg mL{sup -1}. > The biosensor allows sensitive and selective detection of reverse transcriptase in 20 s. - Abstract: Determination of low-abundance human immunodeficiency virus (HIV) enzymes is essential for early detection of the viral infection. Designed for the rapid detection of the virus in human serum, a rapid and ultrasensitive electrochemical assay for HIV-1 reverse transcriptase (HIV-1 RT) is presented in this article. The assay format is based on the formation of a self-assembled monolayer (SAM) of a synthesized ferrocene (Fc)-labeled lipoic acid onto a gold electrode. X-ray photoelectron spectroscopy (XPS) and Time-of-Flight secondary ion mass spectrometry (ToF-SIMS) were employed to confirm the binding of the Fc-labeled lipoic acid to the gold via the Au-S bond. A short RT-specific peptide, VEAIIRILQQLLFIH, was covalently attached to the Fc-labeled lipoic acid. Square wave voltammetry (SWV) offered a two-dimensional measurement of RT based on the anodic shift and reduction of current density of the Fc redox signal upon binding of RT to its specific peptide. This allowed a linear quantification of the target RT in the range of 100-500 pg mL{sup -1}, equivalent to 85.5-427.4 fM, with a detection limit of 50 pg mL{sup -1} (42.7 fM). The developed biosensor is inexpensive, easy to prepare and operate, and allows a highly selective detection of RT in 20 s.

  10. Rapid reversal of human intestinal ischemia-reperfusion induced damage by shedding of injured enterocytes and reepithelialisation.

    Directory of Open Access Journals (Sweden)

    Joep P M Derikx

    Full Text Available BACKGROUND: Intestinal ischemia-reperfusion (IR is a phenomenon related to physiological conditions (e.g. exercise, stress and to pathophysiological events (e.g. acute mesenteric ischemia, aortic surgery. Although intestinal IR has been studied extensively in animals, results remain inconclusive and data on human intestinal IR are scarce. Therefore, an experimental harmless model for human intestinal IR was developed, enabling us to clarify the sequelae of human intestinal IR for the first time. METHODS AND FINDINGS: In 30 patients undergoing pancreatico-duodenectomy we took advantage of the fact that in this procedure a variable length of jejunum is removed. Isolated jejunum (5 cm was subjected to 30 minutes ischemia followed by reperfusion. Intestinal Fatty Acid Binding Protein (I-FABP arteriovenous concentration differences across the bowel segment were measured before and after ischemia to assess epithelial cell damage. Tissue sections were collected after ischemia and at 25, 60 and 120 minutes reperfusion and stained with H&E, and for I-FABP and the apoptosis marker M30. Bonferroni's test was used to compare I-FABP differences. Mean (SEM arteriovenous concentration gradients of I-FABP across the jejunum revealed rapidly developing epithelial cell damage. I-FABP release significantly increased from 290 (46 pg/ml before ischemia towards 3,997 (554 pg/ml immediately after ischemia (p<0.001 and declined gradually to 1,143 (237 pg/ml within 1 hour reperfusion (p<0.001. Directly after ischemia the intestinal epithelial lining was microscopically normal, while subepithelial spaces appeared at the villus tip. However, after 25 minutes reperfusion, enterocyte M30 immunostaining was observed at the villus tip accompanied by shedding of mature enterocytes into the lumen and loss of I-FABP staining. Interestingly, within 60 minutes reperfusion the epithelial barrier resealed, while debris of apoptotic, shedded epithelial cells was observed in the lumen

  11. Aquaponics: What a Way to Grow! Fifth Graders Design Systems for Fish and Plants While Exploring Human Impacts on the Environment

    Science.gov (United States)

    Gillan, Amy; Raja, Shella

    2016-01-01

    In light of increasing populations and dwindling natural resources, elementary teachers play a crucial role in ensuring children understand and commit to more sustainable lifestyles. Climate change, growing pressures on global fisheries, and the harmful effects of traditional agricultural methods exacerbate this call. Coupled with this emphasis is…

  12. Rapid Assessment of Distribution of Wildlife and Human Activities for Prioritizing Conservation Actions in a Patagonian Landscape.

    Science.gov (United States)

    Rivas, Lorena F; Novaro, Andrés J; Funes, Martín C; Walker, R Susan

    2015-01-01

    Large landscapes encompassing reserves and areas with other human uses are necessary for conservation of many species. Generating information for conservation planning over such landscapes may be expensive and time-consuming, though resources for conservation are generally limited and conservation is often urgent. We developed a sign-based occupancy survey to help prioritize conservation interventions by simultaneously assessing the distribution of 3 species, the lesser rhea, guanaco, and mara, and their association with human activities in a 20,000-km2 landscape in the northern Patagonian steppe. We used a single-season occupancy model with spatial rather than temporal replication of surveys in order to reduce costs of multiple visits to sites. We used covariates related to detectability, environmental factors, and different human activities to identify the most plausible models of occupancy, and calculated importance weights of covariates from these models to evaluate relative impacts of human activities on each species. Abundance of goats had the strongest negative association with lesser rheas and guanacos, and road density with maras. With six months of fieldwork, our results provided initial hypotheses for adaptive conservation interventions for each species. Addressing high livestock densities for rheas and guanacos, poaching by urban hunters for all three species, and hunting by rural people for rheas are priorities for conservation in this landscape. Our methodology provided new insights into the responses of these species, although low detection probabilities for maras indicate that the sampling scheme should be altered for future monitoring of this species. This method may be adapted for any large landscape where a rapid, objective means for prioritizing conservation actions on multiple species is needed and data are scarce.

  13. Novel gold nanoparticle trimer reporter probe combined with dry-reagent cotton thread immunoassay device for rapid human ferritin test.

    Science.gov (United States)

    Mao, Xun; Du, Ting-E; Meng, Lili; Song, Tingting

    2015-08-19

    We reported here for the first time on the use of cotton thread combined with novel gold nanoparticle trimer reporter probe for low-cost, sensitive and rapid detection of a lung cancer related biomarker, human ferritin. A model system comprising ferritin as an analyte and a pair of monoclonal antibodies was used to demonstrate the proof-of-concept on the dry-reagent natural cotton thread immunoassay device. Results indicated that the using of novel gold nanoparticle trimer reporter probe greatly improved the sensitivity comparing with traditional gold nanoparticle reporter probe on the cotton thread immunoassay device. The assay avoids multiple incubation and washing steps performed in most conventional protein analyses. Although qualitative tests are realized by observing the color change of the test zone, quantitative data are obtained by recording the optical responses of the test zone with a commercial scanner and corresponding analysis software. Under optimal conditions, the cotton thread immunoassay device was capable of measuring 10 ng/mL human ferritin under room temperature which is sensitive enough for clinical diagnosis. Moreover, the sample solution employed in the assays is just 8 μL, which is much less than traditional lateral flow strip based biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. A novel UHPLC method for the rapid and simultaneous determination of daidzein, genistein and equol in human urine.

    Science.gov (United States)

    Redruello, Begoña; Guadamuro, Lucía; Cuesta, Isabel; Álvarez-Buylla, Jorge R; Mayo, Baltasar; Delgado, Susana

    2015-11-15

    This work reports on a novel method involving reverse-phased ultra-high performance liquid chromatography (UHPLC) plus a spectrophotometric photodiode array/fluorescence (FLR) detection system for determining the concentration of equol and major soy isoflavones (daidzein and genistein) in human urine. The proposed method was validated in terms of its linearity, sensitivity, accuracy (recovery) and precision (intra- and inter-day repeatability). The isoflavone profiles of urine samples from a group of menopausal women following oral soy isoflavone supplementation were determined and compared. Screening for equol-producer status was accomplished with high sensitivity (detection limit of the FLR detector 2.93nM). The method involves a short chromatographic run time compared to conventional HPLC methods while allowing for the simultaneous and reliable quantification of daidzein, genistein and equol in human urine. It also allows for the rapid screening of multiple urine samples when testing for equol production status and checking patient adherence to isoflavone treatment regimens. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Development of a rapid culture method to induce adipocyte differentiation of human bone marrow-derived mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Ninomiya, Yuichi [Translational Research Center, Saitama International Medical, Saitama Medical University, 1397-1 Yamane, Hidaka, Saitama 350-1298 (Japan); Sugahara-Yamashita, Yzumi; Nakachi, Yutaka; Tokuzawa, Yoshimi; Okazaki, Yasushi [Division of Functional Genomics and Systems Medicine, Research Center for Genomic Medicine, Saitama Medical University, Saitama 350-1241 (Japan); Nishiyama, Masahiko, E-mail: yamacho@saitama-med.ac.jp [Translational Research Center, Saitama International Medical, Saitama Medical University, 1397-1 Yamane, Hidaka, Saitama 350-1298 (Japan)

    2010-04-02

    Human mesenchymal stem cells (hMSCs) derived from bone marrow are multipotent stem cells that can regenerate mesenchymal tissues such as adipose, bone or muscle. It is thought that hMSCs can be utilized as a cell resource for tissue engineering and as human models to study cell differentiation mechanisms, such as adipogenesis, osteoblastogenesis and so on. Since it takes 2-3 weeks for hMSCs to differentiate into adipocytes using conventional culture methods, the development of methods to induce faster differentiation into adipocytes is required. In this study we optimized the culture conditions for adipocyte induction to achieve a shorter cultivation time for the induction of adipocyte differentiation in bone marrow-derived hMSCs. Briefly, we used a cocktail of dexamethasone, insulin, methylisobutylxanthine (DIM) plus a peroxisome proliferator-activated receptor {gamma} agonist, rosiglitazone (DIMRo) as a new adipogenic differentiation medium. We successfully shortened the period of cultivation to 7-8 days from 2-3 weeks. We also found that rosiglitazone alone was unable to induce adipocyte differentiation from hMSCs in vitro. However, rosiglitazone appears to enhance hMSC adipogenesis in the presence of other hormones and/or compounds, such as DIM. Furthermore, the inhibitory activity of TGF-{beta}1 on adipogenesis could be investigated using DIMRo-treated hMSCs. We conclude that our rapid new culture method is very useful in measuring the effect of molecules that affect adipogenesis in hMSCs.

  16. Bisphenol A induces a rapid activation of Erk1/2 through GPR30 in human breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Dong, S. [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566 (Japan); Institute of Urban Environment, Chinese Academy of Sciences, Xiamen (China); Terasaka, S. [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566 (Japan); Kiyama, R., E-mail: kiyama.r@aist.go.j [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566 (Japan)

    2011-01-15

    Bisphenol A (BPA) has been considered as an endocrine disruptor due to its ability to interact with estrogen receptors (ERs). While G protein-coupled receptor 30 (GPR30) is a novel estrogen receptor, its role in BPA-induced activation of Erk1/2 remains unknown. Human breast cancer cell lines, MCF-7, MDA-MB-231 and SKBR3, were used as experimental models to discriminate between ERs-dependent, putative ERs-independent and/or GPR30-associated effects. BPA induced a rapid activation of Erk1/2 in both ER{alpha}/{beta}-positive and negative breast cancer cells, and this effect was not blocked with an ER antagonist, ICI 182,780. A small interfering RNA assay revealed that the expression of GPR30 was necessary for BPA-induced activation of Erk1/2 and transcriptional regulation of c-fos. In addition, BPA regulates the expression of c-fos likely through an AP1-mediated pathway. As a conclusion, GPR30 plays an important role in the BPA-induced activation of Erk1/2 in a manner distinguishable from that in ER{alpha}-mediated signaling. - We showed here that the mechanism by which BPA induces the activation of Erk1/2 is distinguishable from the mechanism of ER{alpha}-mediated signaling in human breast cancer cells.

  17. Strategies for rapidly mapping proviral integration sites and assessing cardiogenic potential of nascent human induced pluripotent stem cell clones.

    Science.gov (United States)

    Dambrot, Cheryl; Buermans, Henk P J; Varga, Eszter; Kosmidis, Georgios; Langenberg, Karin; Casini, Simona; Elliott, David A; Dinnyes, Andras; Atsma, Douwe E; Mummery, Christine L; Braam, Stefan R; Davis, Richard P

    2014-10-01

    Recent methodological advances have improved the ease and efficiency of generating human induced pluripotent stem cells (hiPSCs), but this now typically results in a greater number of hiPSC clones being derived than can be wholly characterized. It is therefore imperative that methods are developed which facilitate rapid selection of hiPSC clones most suited for the downstream research aims. Here we describe a combination of procedures enabling the simultaneous screening of multiple clones to determine their genomic integrity as well as their cardiac differentiation potential within two weeks of the putative reprogrammed colonies initially appearing. By coupling splinkerette-PCR with Ion Torrent sequencing, we could ascertain the number and map the proviral integration sites in lentiviral-reprogrammed hiPSCs. In parallel, we developed an effective cardiac differentiation protocol that generated functional cardiomyocytes within 10 days without requiring line-specific optimization for any of the six independent human pluripotent stem cell lines tested. Finally, to demonstrate the scalable potential of these procedures, we picked 20 nascent iPSC clones and performed these independent assays concurrently. Before the clones required passaging, we were able to identify clones with a single integrated copy of the reprogramming vector and robust cardiac differentiation potential for further analysis. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. A Rapid Culture Technique Produces Functional Dendritic-Like Cells from Human Acute Myeloid Leukemia Cell Lines

    Directory of Open Access Journals (Sweden)

    Jian Ning

    2011-01-01

    Full Text Available Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML cells as progenitors from which functional dendritic-like antigen presenting cells (DLC were generated, that constitutively express tumour antigens for recognition by CD8+ T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8+ T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.

  19. Rapid elimination kinetics of free PSA or human kallikrein-related peptidase 2 after initiation of gonadotropin-releasing hormone-antagonist treatment of prostate cancer

    DEFF Research Database (Denmark)

    Ulmert, David; Vickers, Andrew J; Scher, Howard I

    2012-01-01

    The utility of conventional prostate-specific antigen (PSA) measurements in blood for monitoring rapid responses to treatment for prostate cancer is limited because of its slow elimination rate. Prior studies have shown that free PSA (fPSA), intact PSA (iPSA) and human kallikrein-related peptidase...... of tPSA, fPSA, iPSA and hK2 after rapid induction of castration with degarelix (Firmagon(®)), a novel GnRH antagonist....

  20. Human annoyance, acceptability and concern as responses to vibration from the construction of Light Rapid Transit lines in residential environments

    Energy Technology Data Exchange (ETDEWEB)

    Wong-McSweeney, D., E-mail: D.B.C.WongMcSweeney@salford.ac.uk [Acoustics Research Centre, University of Salford, Salford M5 4TW (United Kingdom); Woodcock, J.S.; Peris, E.; Waddington, D.C.; Moorhouse, A.T. [Acoustics Research Centre, University of Salford, Salford M5 4TW (United Kingdom); Redel-Macías, M.D. [Dep. Rural Engineering Campus de Rabanales, University of Córdoba, Córdoba (Spain)

    2016-10-15

    The aim of this paper is to investigate the use of different self-reported measures for assessing the human response to environmental vibration from the construction of an urban LRT (Light Rapid Transit) system. The human response to environmental stressors such as vibration and noise is often expressed in terms of exposure–response relationships that describe annoyance as a function of the magnitude of the vibration. These relationships are often the basis of noise and vibration policy and the setting of limit values. This paper examines measures other than annoyance by expressing exposure–response relationships for vibration in terms of self-reported concern about property damage and acceptability. The exposure–response relationships for concern about property damage and for acceptability are then compared with those for annoyance. It is shown that concern about property damage occurs at vibration levels well below those where there is any risk of damage. Earlier research indicated that concern for damage is an important moderator of the annoyance induced. Acceptability, on the other hand, might be influenced by both annoyance and concern, as well as by other considerations. It is concluded that exposure–response relationships expressing acceptability as a function of vibration exposure could usefully complement existing relationships for annoyance in future policy decisions regarding environmental vibration. The results presented in this paper are derived from data collected through a socio-vibration survey (N = 321) conducted for the construction of an urban LRT in the United Kingdom. - Highlights: • The human response to construction vibration is assessed in residential environments. • Exposure–response relationships are generated based on survey and semi-empirical vibration estimation. • Annoyance, concern and acceptability are compared as response measures. • Concern and acceptability are viable measures complementing annoyance.

  1. Capacitive DNA sensor for rapid and sensitive detection of whole genome human herpesvirus-1 dsDNA in serum.

    Science.gov (United States)

    Cheng, Cheng; Oueslati, Rania; Wu, Jayne; Chen, Jiangang; Eda, Shigetoshi

    2017-06-01

    This work presents a rapid, highly sensitive, low-cost, and specific capacitive DNA sensor for detection of whole genome human herpesvirus-1 DNA. This sensor is capable of direct DNA detection with a response time of 30 s, and it can be used to test standard buffer or serum samples. The sensing approach for DNA detection is based on alternating current (AC) electrokinetics. By applying an inhomogeneous AC electric field on sensor electrodes, positive dielectrophoresis is induced to accelerate DNA hybridization. The same applied AC signal also directly measures the hybridization of target with the probe on the sensor surface. Experiments are conducted to optimize the AC signal, as well as the buffers for probe immobilization and target DNA hybridization. The assay is highly sensitive and specific, with no response to human herpesvirus-2 DNA at 5 ng/mL and a LOD of 1.0 pg/mL (6.5 copies/μL or 10.7 aM) in standard buffer. When testing the double stranded (ds) DNA spiked in human serum samples, the sensor yields a LOD of 20.0 pg/mL (129.5 copies/μL or 0.21 femtomolar (fM)) in neat serum. In this work, the target is whole genome dsDNA, consequently the test can be performed without the use of enzyme or amplification, which considerably simplifies the sensor operation and is highly suitable for point of care disease diagnosis. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Rapid effects of 17beta-estradiol on epithelial TRPV6 Ca2+ channel in human T84 colonic cells.

    LENUS (Irish Health Repository)

    Irnaten, Mustapha

    2008-11-01

    The control of calcium homeostasis is essential for cell survival and is of crucial importance for several physiological functions. The discovery of the epithelial calcium channel Transient Receptor Potential Vaniloid (TRPV6) in intestine has uncovered important Ca(2+) absorptive pathways involved in the regulation of whole body Ca(2+) homeostasis. The role of steroid hormone 17beta-estradiol (E(2)), in [Ca(2+)](i) regulation involving TRPV6 has been only limited at the protein expression levels in over-expressing heterologous systems. In the present study, using a combination of calcium-imaging, whole-cell patch-clamp techniques and siRNA technology to specifically knockdown TRPV6 protein expression, we were able to (i) show that TRPV6 is natively, rather than exogenously, expressed at mRNA and protein levels in human T84 colonic cells, (ii) characterize functional TRPV6 channels and (iii) demonstrate, for the first time, the rapid effects of E(2) in [Ca(2+)](i) regulation involving directly TRPV6 channels in T84 cells. Treatment with E(2) rapidly (<5 min) enhanced [Ca(2+)](i) and this increase was partially but significantly prevented when cells were pre-treated with ruthenium red and completely abolished in cells treated with siRNA specifically targeting TRPV6 protein expression. These results indicate that when cells are stimulated by E(2), Ca(2+) enters the cell through TRPV6 channels. TRPV6 channels in T84 cells contribute to the Ca(2+) entry\\/signalling pathway that is sensitive to 17beta-estradiol.

  3. Performance of a rapid, on-site human immunodeficiency virus antibody assay in a public health setting.

    Science.gov (United States)

    Kassler, W J; Haley, C; Jones, W K; Gerber, A R; Kennedy, E J; George, J R

    1995-11-01

    Rapid, on-site human immunodeficiency virus (HIV) testing has the potential to improve the delivery of prevention services in publicly funded counseling and testing sites. The Single Use Diagnostic System (SUDS) HIV-1 is the only rapid enzyme immunoassay (EIA) approved for diagnostic use in the United States. To evaluate the feasibility of using SUDS in public clinics and to validate the test's performance in a public health laboratory, we conducted blinded SUDS testing on plasma sent for HIV testing. From 19 March through 30 June 1993, 1,923 consecutive samples from a sexually transmitted diseases clinic and an HIV counseling and testing clinic were tested on site with SUDS. Tests done in the first two weeks with a malfunctioning centrifuge n = 402) and those done when there were excessively high temperatures in the laboratory (n = 53) were analyzed separately. Of 1,466 tests, 39 were positive by both SUDS and EIA (with Western blot [immunoblot] confirmation) and 7 were SUDS positive and EIA negative. Western blotting was used as the "gold standard" to adjudicate these discrepancies. There were no SUDS-negative and EIA-positive tests. Compared with that of EIA (with Western blot confirmation), the sensitivity of SUDS was 100% (95% confidence interval, 88.8 to 100%) and the specificity was 99.5% (95% confidence interval, 98.9 to 99.8%). The positive predictive value of SUDS was 88% in the STD clinic and 81% in the HIV counseling and testing clinic. There was a 7.7-fold increase in false positives, from 0.48 to 3.7%, when there was inadequate centrifugation and when the temperature exceeded the manufacturer's recommendations. Rapid, on-site HIV testing by the SUDS assay is feasible and practical in public health settings. The test can be performed accurately, at reasonable cost, and within the time frame of a typical clinic visit. Caution should be used, however, as two conditions adversely affected the accuracy of this test: inadequate specimen preparation and

  4. Rapid spread of the invasive yellow-legged hornet in France: the role of human-mediated dispersal and the effects of control measures

    OpenAIRE

    Robinet, Christelle; Suppo, Christelle; Darrouzet, Eric

    2017-01-01

    1.The invasive yellow-legged hornet was first discovered in Europe, in south-western France, in 2004. It has since spread very rapidly and has caused significant mortality among honey bees and native entomofauna. It also poses a risk to humans because its sting provokes allergic reactions. The objectives of this study were the following: (i) to disentangle the roles played by human-mediated dispersal and self-mediated dispersal in the species’ rapid range expansion and (ii) to estimate the in...

  5. Melting ice, growing trade?

    National Research Council Canada - National Science Library

    Sami Bensassi; Julienne C. Stroeve; Inmaculada Martínez-Zarzoso; Andrew P. Barrett

    2016-01-01

    Abstract Large reductions in Arctic sea ice, most notably in summer, coupled with growing interest in Arctic shipping and resource exploitation have renewed interest in the economic potential of the Northern Sea Route (NSR...

  6. Rapid detection and typing of live bacteria from human joint fluid samples by utilizing an integrated microfluidic system.

    Science.gov (United States)

    Chang, Wen-Hsin; Wang, Chih-Hung; Lin, Chih-Lin; Wu, Jiunn-Jong; Lee, Mel S; Lee, Gwo-Bin

    2015-04-15

    Periprosthetic joint infection (PJI) is one of the most dreading complications that hinder the merits of an arthroplasty. A prerequisite for treatment of the above procedure is rapid detection of live bacteria to prevent its recurrence and proper choice of antibiotics. Conventional culture methods are time-consuming and associated with a high false negative rate. Amplification of bacterial genetic materials requires a tedious process but is associated with a high false positive rate. An integrated microfluidic system capable of molecular diagnosis for detecting live bacteria was reported in our previous work. However, the system could not provide detailed information about infectious bacteria for the subsequent antibiotic choices. Furthermore, it took at least 55min to finish the entire process. In this work, a microfluidic platform using ethidium monoazide (EMA) which can only penetrate into dead bacteria is presented for live bacteria detection and typing within a short period of time (30min for the detection of live bacteria and another 40min for the typing of bacteria strains). We tested the proposed system by using human joint fluid samples and found its limit of detection for bacterial detection equal to 10(2)CFU (colony formation unit) for live bacteria detection with gold nanoparticle probes and 10(2)-10(4)CFU for typing bacteria by an on-chip polymerase chain reaction. The whole procedure of the integrated microfluidic system is automated with little human intervention. Moreover, this is the first time that sequential live bacteria detection and typing are demonstrated on the same microfluidic platform. Based on the promising results, the proposed system may become in the near future an auxiliary tool for immediate medical decision and choice of antibiotics in routine arthroplasties or PJI's. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Rapid replacement of prevailing genotype of human respiratory syncytial virus by genotype ON1 in Beijing, 2012-2014.

    Science.gov (United States)

    Cui, Guanglin; Zhu, Runan; Deng, Jie; Zhao, Linqing; Sun, Yu; Wang, Fang; Qian, Yuan

    2015-07-01

    Human respiratory syncytial virus (HRSV) is the most common viral pathogen causing lower respiratory infections in infants and young children worldwide. HRSV ON1 genotype in subgroup A with a characteristic of a 72 nucleotide duplication in the second highly variable region of attachment glycoprotein gene, has been reported in some countries since it was first detected in clinical samples collected in Canada in 2010. In this study, 557 HRSV antigen-positive nasopharyngeal aspirates were randomly selected during 2012/2013 to 2013/2014 HRSV seasons in Beijing for subgroup typing and for ON1 genotype screening by using a PCR based method developed for easily identifying genotype ON1 out of strains of subtype A. It was found that subgroup B was dominant in the 2012/2013 season and sudden shift of subgroup dominance from B to A and rapid replacement of previously prevailing NA1 genotype by ON1 genotype occurred in the 2013/2014 season. Reversible amino acid replacement in the G protein gene was found in a new branch of ON1 genotype. The evolutionary rate of the 351 global ON1 sequences was estimated to 7.34 × 10(-3) nucleotide substitutions per site per year (95% highest probability density intervals, HPD, 5.71 × 10(-3) to 9.04 × 10(-3)), with the time of most recent common ancestor dating back to June 2009. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. LC-ESI/MS/MS method for rapid screening and confirmation of 44 exogenous anabolic steroids in human urine.

    Science.gov (United States)

    Jeon, Byoung Wook; Yoo, Hye Hyun; Jeong, Eun Sook; Kim, Ho Jun; Jin, Changbae; Kim, Dong Hyun; Lee, Jaeick

    2011-09-01

    A sensitive and rapid method based on liquid chromatography-triple-quadrupole tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI) has been developed and validated for the screening and confirmation of 44 exogenous anabolic steroids (29 parent steroids and 15 metabolites) in human urine. The method involves an enzymatic hydrolysis, liquid-liquid extraction, and detection by LC-MS/MS. A triple-quadrupole mass spectrometer was operated in positive ESI mode with selected reaction monitoring (SRM) mode for the screening and product ion scan mode for the confirmation. The protonated molecular ions were used as precursor ions for the SRM analysis and product ion scan. The intraday and interday precisions of the target analytes at concentrations of the minimum required performance levels for the screening were 2-14% and 2-15%, respectively. The limits of detection for the screening and confirmation method were 0.1-10 ng/mL and 0.2-10 ng/mL, respectively, for 44 steroids. This method was successfully applied to analysis of urine samples from suspected anabolic steroid abusers.

  9. A Rapid and Sensitive HPLC-Fluorescence Method for Determination of Mirtazapine and Its two Major Metabolites in Human Plasma.

    Science.gov (United States)

    Lavasani, Hoda; Giorgi, Mario; Sheikholeslami, Behjat; Hedayati, Mohammadhasan; Rouini, Mohammad Reza

    2014-01-01

    A rapid and sensitive HPLC method has been developed for the quantification of mirtazapine (MRZ), a noradrenergic and specific serotonergic inhibitor antidepressant (NaSSA) and its two major metabolites N-desmethyl mirtazapine (NDM) and 8-hydroxymirtazapine (8-OHM) in human plasma. The separation was achieved using Chromolith C18 column and a mobile phase of acetonitrile: phosphate buffer (pH = 3, 20:80, v/v) in isocratic mode at a flow rate of 2 mL/min. A fluorescence detector was set at 290 and 350 nm for excitation and emission, respectively. Zolpidem was used as the internal standard. Liquid-liquid extraction was applied for sample clean up. All analytes were eluted in less than 5 minutes with LOQ of 1 ng/mL for MRZ and 2 ng/mL for both NDM and 8-OHM. The developed method was successfully applied to quantify MRZ and its metabolites in plasma of a healthy volunteer.

  10. A New Surface Plasmon Resonance-Based Immunoassay for Rapid, Reproducible and Sensitive Quantification of Pentraxin-3 in Human Plasma

    Directory of Open Access Journals (Sweden)

    Mara Canovi

    2014-06-01

    Full Text Available A new immunoassay based on surface plasmon resonance (SPR for the rapid, reproducible and sensitive determination of pentraxin-3 (PTX3 levels in human plasma has been developed and characterized. The method involves a 3-min flow of plasma over a sensor chip pre-coated with a monoclonal anti-PTX3 antibody (MNB4, followed by a 3-min flow of a polyclonal anti-PTX3 antibody (pAb, required for specific recognition of captured PTX3. The SPR signal generated with this secondary antibody linearly correlates with the plasma PTX3 concentration, in the range of 5–1500 ng/mL, with a lowest limit of detection of 5 ng/mL. The PTX3 concentrations determined with the SPR-based immunoassay in the plasma of 21 patients with sepsis, ranging 15–1600 ng/mL, were superimposable to those found in a classic ELISA immunoassay. Since the PTX3 concentration in the plasma of healthy subjects is <2 ng/mL, but markedly rises in certain medical conditions, the method is useful to quantify pathological levels of this important biomarker, with important diagnostic applications. In comparison with the classic ELISA, the SPR-based approach is much faster (30 min versus 4–5 h and could be exploited for the development of new cost-effective SPR devices for point-of-care diagnosis.

  11. Rapid and sensitive liquid chromatography tandem mass spectrometry method for the quantification of ambroxol in human plasma.

    Science.gov (United States)

    Hu, Wanqun; Xu, Yu; Liu, Fei; Liu, Aixiang; Guo, Qingxiang

    2008-10-01

    A sensitive, specific and rapid high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was described and validated for the quantification of ambroxol in human plasma using enalaprilat as the internal standard (IS). Chromatographic separation was performed on a Lichrospher CN column with a mobile phase of methanol and water (containing 0.1% formic acid) (70:30, v/v). The total run time was 5.0 min for each sample. The analytes was detected by mass spectrometry with electrospray ionization source in positive selected reaction monitoring mode. The precursor-fragment ion reaction for ambroxol was m/z 378.9 --> 263.8, and for IS was m/z 349.0 --> 205.9. The linearity was established over the concentration range of 1.56-400.00 ng/mL. The inter-day and the intra-day precisions were all within 10%. A simple protein precipitation with methanol was adopted for sample preparation. The extraction recoveries of ambroxol and IS were higher than 90.80%. The validated method was successfully applied in pharmacokinetic study after oral administration of 90 mg ambroxol to 24 healthy volunteers.

  12. Rapid genotyping of carcinogenic human papillomavirus by loop-mediated isothermal amplification using a new automated DNA test (Clinichip HPV™).

    Science.gov (United States)

    Satoh, Toyomi; Matsumoto, Koji; Fujii, Takuma; Sato, Osamu; Gemma, Nobuhiro; Onuki, Mamiko; Saito, Hiroshi; Aoki, Daisuke; Hirai, Yasuo; Yoshikawa, Hiroyuki

    2013-03-01

    This study was designed to evaluate the Clinichip HPV test, a new DNA test that detects carcinogenic human papillomavirus (HPV) rapidly by loop-mediated isothermal amplification and performs genotyping of all 13 carcinogenic types using automated DNA chip technology with an assay time 2.5h. Using this test, 247 Japanese women (109 with normal cytology, 43 with cervical intraepithelial neoplasia grade 1, 60 with cervical intraepithelial neoplasia grade 2/3 and 35 with invasive cervical cancer) were tested for carcinogenic HPV genotypes. The results were compared to those obtained by the polymerase chain reaction-amplified DNA sequencing using 13 type-specific primers. Overall, there was very good agreement for the detection of carcinogenic HPV between the Clinichip test and direct sequencing, with 95.5% total agreement and a kappa value of 0.91. Comparison of the detection of individual HPV types shows that the overall agreement was also high (range: 96.8-100%). In women with cervical intraepithelial neoplasia grade 2 or worse, the detection rate of carcinogenic HPV was 95.7% by both the Clinichip test and the direct-sequencing method, indicating complete agreement between the two methods. In conclusion, it was found that the Clinichip test is a promising new laboratory method for genotyping of carcinogenic HPV. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Rapid analysis of the interactions between drugs and human serum albumin (HSA) using high-performance affinity chromatography (HPAC).

    Science.gov (United States)

    Kim, Hee Seung; Wainer, Irving W

    2008-07-01

    This study used a combination of zonal elution and frontal affinity chromatography on immobilized human serum albumin (HSA) high-performance affinity chromatography (HPAC) column to examine the association constants of various compounds that have been studied by equilibrium dialysis or ultra filtration. A standard plot was generated from retention factors of reference compounds using zonal elution chromatography against association constants of reference compounds using frontal affinity chromatography. The linear relationship was established (r2=0.9993) between retention factors and association constants of reference compounds. This standard plot was later used for rapid determination of association constants of various drugs which show low to medium binding affinity to HSA. Association constants of those drugs from this study were compared to that of more generally used methods (i.e., equilibrium dialysis or ultra filtration) from literature and resulted in a relatively high correlation (r2=0.945) value. This combination of zonal elution and frontal affinity chromatography method for determining association constants showed several advantages against traditional methods. Depending on drugs of interest, an association constant of drug to HSA can be measured as fast as 1.5 min. Other notable advantages include an ease of automation and its ability to distinguish association constants of chiral compounds at the same time. The same approach could be used for studying interaction of other drugs and proteins and should further improve overall drug screening process.

  14. Measurement of proliferation and disappearance of rapid turnover cell populations in human studies using deuterium-labeled glucose.

    Science.gov (United States)

    Macallan, Derek C; Asquith, Becca; Zhang, Yan; de Lara, Catherine; Ghattas, Hala; Defoiche, Julien; Beverley, Peter C L

    2009-01-01

    Cell proliferation may be measured in vivo by quantifying DNA synthesis with isotopically labeled deoxyribonucleotide precursors. Deuterium-labeled glucose is one such precursor which, because it achieves high levels of enrichment for a short period, is well suited to the study of rapidly dividing cells, in contrast to the longer term labeling achieved with heavy water ((2)H(2)O). As deuterium is non-radioactive and glucose can be readily administered, this approach is suitable for clinical studies. It has been widely applied to investigate human lymphocyte proliferation, but solid tissue samples may also be analyzed. Rate, duration and route (intravenous or oral) of [6,6-(2)H(2)]-glucose administration should be adapted to the target cell of interest. For lymphocytes, cell separation is best achieved by fluorescence activated cell sorting (FACS), although magnetic bead separation is an alternative. DNA is then extracted, hydrolyzed enzymatically and analyzed by gas chromatography mass spectrometry (GC/MS). Appropriate mathematical modeling is critical to interpretation. Typical time requirements are as follows: labeling, 10-24 h; sampling, approximately 3 weeks; DNA extraction/derivatization, 2-3 d; and GC/MS analysis, approximately 2 d.

  15. Rapid and efficient conversion of integration-free human induced pluripotent stem cells to GMP-grade culture conditions.

    Directory of Open Access Journals (Sweden)

    Jens Durruthy-Durruthy

    Full Text Available Data suggest that clinical applications of human induced pluripotent stem cells (hiPSCs will be realized. Nonetheless, clinical applications will require hiPSCs that are free of exogenous DNA and that can be manufactured through Good Manufacturing Practice (GMP. Optimally, derivation of hiPSCs should be rapid and efficient in order to minimize manipulations, reduce potential for accumulation of mutations and minimize financial costs. Previous studies reported the use of modified synthetic mRNAs to reprogram fibroblasts to a pluripotent state. Here, we provide an optimized, fully chemically defined and feeder-free protocol for the derivation of hiPSCs using synthetic mRNAs. The protocol results in derivation of fully reprogrammed hiPSC lines from adult dermal fibroblasts in less than two weeks. The hiPSC lines were successfully tested for their identity, purity, stability and safety at a GMP facility and cryopreserved. To our knowledge, as a proof of principle, these are the first integration-free iPSCs lines that were reproducibly generated through synthetic mRNA reprogramming that could be putatively used for clinical purposes.

  16. Protecting drinking water: Rapid detection of human fecal contamination, injured and non-culturable pathogenic microbes in water systems

    Energy Technology Data Exchange (ETDEWEB)

    White, D.C.; Nivens, D.E.; Arrage, A.A.; Appelgate, B.M.; Reardon, S.R.; Sayler, G.S.

    1996-05-01

    The rapid, potentially-automatable extraction of filter retentates has allowed quantitative detection of the unique biomarker for human fecal contamination, coprostanol, and the signature lipid biomarkers for total cellular biomass, viable cellular biomass, lipopolysaccharide (endotoxin). This method may be integrated with DNA based gene probe analysis for specific strains and enzyme activities. Not only does the analysis provide for detection of injured and non-culturable microbes but it also provides biomarkers characteristic of microbes exposed to biocides and disinfectants that can be utilized to monitor effectiveness of water mitigation/treatment. The analysis schemes involve filtration of the water or direct extraction of biofilms in sidestream chambers, supercritical fluid and/or liquid extraction, derivatization, and analysis of ``signature`` patterns by gas chromatography/mass spectrometry. Signature lipid biomarkers of interest are diglycerides, steroids including coprostanol and its isomers, poly-{beta}- hydroxyalcanoates (PHA), phospholipid ester-linked fatty acids (PLFA), and the lipopolysaccharide lipid A hydroxy fatty acids. PLFA found in polar lipid fractions estimate total viable cellular biomass, whereas the total cellular biomass can be calculated from diglyceride/phospholipid ester-linked fatty acids ratios. Furthermore, direct evidence of mitigation/treatment effectiveness can be ascertained by detection of diglycerides, respiratory quinones, PHA, and PLFA markers indicative of metabolic stress and toxicity such as trans monoenoic PLFA as well as oxirane and dicarboxylic fatty acids derived from the PLFA.

  17. Rapid detection of herpes simplex virus in clinical specimens with human embryonic lung fibroblast and primary rabbit kidney cell cultures.

    Science.gov (United States)

    Callihan, D R; Menegus, M A

    1984-04-01

    The performance of a culture system for isolation of herpes simplex virus, consisting of one tube each of human embryonic lung fibroblasts and primary rabbit kidney cells, was evaluated. Cultures were incubated at 37 degrees C on a roller drum and observed daily for characteristic cytopathic effect for 5 days. During 1982, a positive isolation rate of 28.1% was seen among 3,154 specimens submitted. Cultures from genital sources were positive more frequently from males (43.8%) than from females (25.5%). Oral lesion cultures were positive as often from males (34.6%) as from females (38.4%). Although detection of herpes simplex virus occurred significantly earlier in rabbit kidney cells on days 1 and 2 of incubation, by day 3 the number of positive cultures was nearly the same in both cell types. By day 4 of incubation, 99.5% of the positive cultures were detected. These results demonstrate that cell culture can be a rapid and sensitive method for detecting herpes simplex virus.

  18. A rapid and simple determination of A77 1726 in human serum by high-performance liquid chromatography and its application for optimization of leflunomide therapy

    NARCIS (Netherlands)

    van Roon, EN; Yska, JP; Raemaekers, J; Jansen, TLTA; van Wanrooy, M; Brouwers, JRBJ

    2004-01-01

    Leflunomide is a disease-modifying antirheumatic drug, which is bioactivated by fort-nation of A77 1726. In this study a rapid and simple quantitative assay using a reversed phase HPLC-UV method is validated for detection of A77 1726 in human serum. The HPLC-UV method uses a mobile phase consisting

  19. Rapid yeast estrogen bioassays stably expressing human estrogen receptors alpha and beta, and green fluorescent protein: a comparison of different compounds on both receptor types

    NARCIS (Netherlands)

    Bovee, T.F.H.; Helsdingen, J.R.; Rietjens, I.M.C.M.; Keijer, J.; Hoogenboom, L.A.P.

    2004-01-01

    Previously, we described the construction of a rapid yeast bioassay stably expressing human estrogen receptor (hER) and yeast enhanced green fluorescent protein (yEGFP) in response to estrogens. In the present study, the properties of this assay were further studied by testing a series of estrogenic

  20. The role of rapid testing and clinical decision in the diagnosis of human influenza A H1N1 infection.

    Science.gov (United States)

    BinSaeed, Abdulaziz A; Siddiqui, Amna R; Mandil, Ahmed M; Torchyan, Armen A; Tayel, Salwa A; Shaikh, Shaffi A; Habib, Hanan A; Al-Khattaf, Abdulaziz S

    2014-03-01

    To evaluate the role of the rapid influenza diagnostic test (RIDT) and clinical decision in the diagnosis of H1N1. In November 2009, 290 suspected influenza patients were examined for H1N1 during an outbreak in Riyadh, Saudi Arabia. Nasopharyngeal swabs were analyzed using Directigen EZ Flu A+B kit. Monoclonal anti-human influenza A/B and reverse transcription- polymerase chain reaction (RT-PCR) were used. Positive and negative controls were used in each run of specimens. Validity indices were calculated for RIDT and clinical diagnostic criteria. The sensitivity and specificity of RIDT were 40.5% (95% confidence interval [CI]: 33.0-48.5), and 94.5% (95% CI: 88.6-97.6). The sensitivity of clinical decision was 66.3% (95% CI: 58.4-73.4), and the specificity was 65.4% (95% CI: 56.3-73.4). The sensitivity of clinical decision was higher in early presenters (79.2%; 95% CI: 57.3-92.1). The RIDT sensitivity was higher in younger patients (48.4%; 95% CI: 35.7-61.3). The positive predictive value (PPV) was 90.4% (95% CI: 80.7-95.7) for RIDT, and 71.1% (95% CI: 63.1-78.0) for clinical decision. The PPV for RIDT was greater for older (94.7%; 95% CI: 80.9-99.1) and late (90.7%; 95% CI: 76.9-97.0) presenters. The adjusted odds ratio for clinical decision was significant for cough, headache, and fatigue. The RIDT can be useful in epidemics and high prevalence areas, whereas clinical decision, and RT-PCR complement the diagnosis of H1N1 in any setting.

  1. Antioxidants cause rapid expansion of human adipose-derived mesenchymal stem cells via CDK and CDK inhibitor regulation

    Science.gov (United States)

    2013-01-01

    Background Antioxidants have been shown to enhance the proliferation of adipose-derived mesenchymal stem cells (ADMSCs) in vitro, although the detailed mechanism(s) and potential side effects are not fully understood. In this study, human ADMSCs cultured in ImF-A medium supplemented with antioxidants (N-acetyl-l-cysteine and ascorbic acid-2-phosphate) and fibroblast growth factor 2 (FGF-2) were compared with ADMSCs cultured with FGF-2 alone (ImF) or with FGF-2 under 5% pO2 conditions (ImF-H). Results During log-phase growth, exposure to ImF-A resulted in a higher percentage of ADMSCs in the S phase of the cell cycle and a smaller percentage in G0/G1 phase. This resulted in a significantly reduced cell-doubling time and increased number of cells in the antioxidant-supplemented cultures compared with those supplemented with FGF-2 alone, an approximately 225% higher cell density after 7 days. Western blotting showed that the levels of the CDK inhibitors p21 and p27 decreased after ImF-A treatment, whereas CDK2, CDK4, and CDC2 levels clearly increased. In addition, ImF-A resulted in significant reduction in the expression of CD29, CD90, and CD105, whereas relative telomere length, osteogenesis, adipogenesis, and chondrogenesis were enhanced. The results were similar for ADMSCs treated with antioxidants and those under hypoxic conditions. Conclusion Antioxidant treatment promotes entry of ADMSCs into the S phase by suppressing cyclin-dependent kinase inhibitors and results in rapid cell proliferation similar to that observed under hypoxic conditions. PMID:23915242

  2. NDER: A novel web application using annotated whole slide images for rapid improvements in human pattern recognition

    Directory of Open Access Journals (Sweden)

    Nicholas P Reder

    2016-01-01

    Full Text Available Context: Whole-slide images (WSIs present a rich source of information for education, training, and quality assurance. However, they are often used in a fashion similar to glass slides rather than in novel ways that leverage the advantages of WSI. We have created a pipeline to transform annotated WSI into pattern recognition training, and quality assurance web application called novel diagnostic electronic resource (NDER. Aims: Create an efficient workflow for extracting annotated WSI for use by NDER, an attractive web application that provides high-throughput training. Materials and Methods: WSI were annotated by a resident and classified into five categories. Two methods of extracting images and creating image databases were compared. Extraction Method 1: Manual extraction of still images and validation of each image by four breast pathologists. Extraction Method 2: Validation of annotated regions on the WSI by a single experienced breast pathologist and automated extraction of still images tagged by diagnosis. The extracted still images were used by NDER. NDER briefly displays an image, requires users to classify the image after time has expired, then gives users immediate feedback. Results: The NDER workflow is efficient: annotation of a WSI requires 5 min and validation by an expert pathologist requires An additional one to 2 min. The pipeline is highly automated, with only annotation and validation requiring human input. NDER effectively displays hundreds of high-quality, high-resolution images and provides immediate feedback to users during a 30 min session. Conclusions: NDER efficiently uses annotated WSI to rapidly increase pattern recognition and evaluate for diagnostic proficiency.

  3. [Studies on the alkaloids of Senecio scandens growing in Guangdong].

    Science.gov (United States)

    Guo, Xiao-Fang; Liu, Meng-Hua; Peng, Wei; Wang, Yong-Gang; Yang, Cui-Ping; Su, Wei-Wei

    2011-05-01

    To study alkaloids of Senecio scandens growing in Guangdong. The rapid resolution liquid chromatography-electrospray ionization mass spectrometry (RRLC-ESI-MS/MS) was used to analyse alkaloids of Senecio scandens growing in Guangdong, and senkirkine was isolated and purified by silica gel column chromatography. Four alkaloids were identified as senkirkine, dehydrosenkirkine, monocrotaline and adonifoline, and senkirkine was firstly isolated from Senecio scandens growing in Guangdong. Senkirkine is the main component of Senecio scandens growing in Guangdong.

  4. Rapid insulinotropic action of low doses of bisphenol-A on mouse and human islets of Langerhans: role of estrogen receptor β.

    Directory of Open Access Journals (Sweden)

    Sergi Soriano

    Full Text Available Bisphenol-A (BPA is a widespread endocrine-disrupting chemical (EDC used as the base compound in the manufacture of polycarbonate plastics. It alters pancreatic β-cell function and can be considered a risk factor for type 2 diabetes in rodents. Here we used ERβ-/- mice to study whether ERβ is involved in the rapid regulation of K(ATP channel activity, calcium signals and insulin release elicited by environmentally relevant doses of BPA (1 nM. We also investigated these effects of BPA in β-cells and whole islets of Langerhans from humans. 1 nM BPA rapidly decreased K(ATP channel activity, increased glucose-induced [Ca(2+](i signals and insulin release in β-cells from WT mice but not in cells from ERβ-/- mice. The rapid reduction in the K(ATP channel activity and the insulinotropic effect was seen in human cells and islets. BPA actions were stronger in human islets compared to mouse islets when the same BPA concentration was used. Our findings suggest that BPA behaves as a strong estrogen via nuclear ERβ and indicate that results obtained with BPA in mouse β-cells may be extrapolated to humans. This supports that BPA should be considered as a risk factor for metabolic disorders in humans.

  5. Growing skull fracture in a red-tailed hawk (Buteo jamaicensis).

    Science.gov (United States)

    Rush, E Marie; Shores, Andrew; Meintel, Sarah; Hathcock, John T

    2014-09-01

    Growing skull fractures have been reported in humans for many years, usually resulting from injury to the soft skull during the rapid growth period of an infant's life. Nestling raptors have thin, fragile skulls, a rapid growth rate, and compete aggressively for food items. Skull trauma may occur, which may lead to the development of a growing skull fracture. Growing skull fractures may be under-diagnosed in raptor rehabilitation facilities that do not have access to advanced technologic equipment. Three-dimensional (3-D) computed tomography was used to diagnose a growing skull fracture in a red-tailed hawk (Buteo jamaicensis). The lesion was surgically repaired and the animal was eventually returned to the wild. This is the first report of a growing skull fracture in an animal. In this case, 3-D computed topographic imaging was utilized to diagnose a growing skull fracture in a red-tailed hawk, surgical repair was performed, and the bird recovered completely and was ultimately released.

  6. Growing Plants and Minds

    Science.gov (United States)

    Presser, Ashley Lewis; Kamdar, Danae; Vidiksis, Regan; Goldstein, Marion; Dominguez, Ximena; Orr, Jillian

    2017-01-01

    Many preschool classrooms explore plant growth. However, because many plants take a long time to grow, it is often hard to facilitate engagement in some practices (i.e., since change is typically not observable from one day to another, children often forget their prior predictions or cannot recall what plants looked like days or weeks earlier).…

  7. Growing Up with "1984."

    Science.gov (United States)

    Franza, August

    1983-01-01

    Relates changing student reaction to George Orwell's "1984" over 20 years of teaching. Finds present high school students' acceptance of Orwell's bleak world vision both a sign of student honesty and a frightening indication of the growing reality of the book. (MM)

  8. Growing through Literature.

    Science.gov (United States)

    Thomas, Barbara J.

    "Growing through Literature" is a curriculum using Joan M. and Erik H. Erikson's theory of the Life Cycle as a structure for selecting and teaching literature to inner-city high school students at Brighton High School in Massachusetts. The program consists of four component parts: Journals, Selected Stories, Discussion, and…

  9. Growing Old in Exile

    DEFF Research Database (Denmark)

    Liversage, Anika; Mirdal, Gretty Mizrahi

    2017-01-01

    Some studies on immigrants and ageing focus on the question of return; others focus on how immigrants, who grow old in their countries of destination, ‘age in place’, including whether they turn to their children or to public host country provisions for care and support. However, the issues of re...

  10. Rapid expression of an anti-human C5 chimeric Fab utilizing a vector that replicates in COS and 293 cells.

    Science.gov (United States)

    Evans, M J; Hartman, S L; Wolff, D W; Rollins, S A; Squinto, S P

    1995-07-17

    Inhibition of complement system activation requires the development of soluble nonimmunogenic inhibitors with good tissue penetrating abilities that are themselves unable to activate complement. Chimeric mouse/human Fabs capable of blocking the activity of complement proteins are likely to fulfill these criteria. Several monoclonal antibodies that inhibit the activation of the human complement system have recently been developed. To examine the properties of chimeric Fab derived from these monoclonal antibodies, we have developed an expression system which allows the rapid production of milligram quantities of chimeric Fab. Both the chimeric light chain and the chimeric Fd were co-expressed from the same vector, pAPEX-3P. This vector contains the SV40 origin of replication, which allows the rapid production of chimeric Fab in COS cells for preliminary characterization. Additionally, pAPEX-3P contains the Epstein-Barr virus origin of replication and a puromycin selectable marker for maintenance as a stable episome in human cell lines. A production system consisting of transfected 293-EBNA cells cultured in serum free medium followed by protein G-Sepharose chromatography of the conditioned medium was found to be sufficient for the rapid production of purified chimeric Fab. Here we have utilized this expression system to demonstrate that an anti-human C5 chimeric Fab was a potent inhibitor of complement activation in both in vitro activation assays and an ex vivo model of complement-mediated tissue damage.

  11. Rapid engraftment by peripheral blood progenitor cells mobilized by recombinant human stem cell factor and recombinant human granulocyte colony-stimulating factor in nonhuman primates.

    Science.gov (United States)

    Andrews, R G; Briddell, R A; Knitter, G H; Rowley, S D; Appelbaum, F R; McNiece, I K

    1995-01-01

    We have previously shown that administration of low-dose recombinant human stem cell factor (rhSCF) plus recombinant human granulocyte colony-stimulating factor (rhG-CSF) to baboons mobilizes greater numbers of progenitor cells in the blood than does administration of rhG-CSF alone. The purpose of the present study was to determine whether marrow repopulating cells are present in the blood of nonhuman primates administered low-dose rhSCF plus rhG-CSF, and if present, whether these cells engraft lethally irradiated recipients as rapidly as blood cells mobilized by treatment with rhG-CSF alone. One group of baboons was administered low-dose rhSCF (25 micrograms/kg/d) plus rhG-CSF (100 micrograms/kg/d) while a second group received rhG-CSF alone (100 micrograms/kg/d). Each animal underwent a single 2-hour leukapheresis occurring the day when the number of progenitor cells per volume of blood was maximal. For baboons administered low-dose rhSCF plus rhG-CSF, the leukapheresis products contained 1.8-fold more mononuclear cells and 14.0-fold more progenitor cells compared to the leukapheresis products from animals treated with rhG-CSF alone. All animals successfully engrafted after transplantation of cryopreserved autologous blood cells. In animals transplanted with low-dose rhSCF plus rhG-CSF mobilized blood cells, we observed a time to a platelet count of > 20,000 was 8 days +/- 0, to a white blood cell count (WBC) of > 1,000 was 11 +/- 1 days, and to an absolute neutrophil count (ANC) of > 500 was 12 +/- 1 days. These results compared with 42 +/- 12, 16 +/- 1, and 24 +/- 4 days to achieve platelets > 20,000, WBC > 1,000, and ANC > 500, respectively, for baboons transplanted with rhG-CSF mobilized blood cells. Animals transplanted with low-dose rhSCF plus rhG-CSF mobilized blood cells had blood counts equivalent to pretransplant values within 3 weeks after transplant. The results suggest that the combination of low-dose rhSCF plus rhG-CSF mobilizes greater numbers of

  12. A rapid crosstalk of human gammadelta T cells and monocytes drives the acute inflammation in bacterial infections.

    Directory of Open Access Journals (Sweden)

    Matthias Eberl

    2009-02-01

    Full Text Available Vgamma9/Vdelta2 T cells are a minor subset of T cells in human blood and differ from other T cells by their immediate responsiveness to microbes. We previously demonstrated that the primary target for Vgamma9/Vdelta2 T cells is (E-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP, an essential metabolite produced by a large range of pathogens. Here we wished to study the consequence of this unique responsiveness in microbial infection. The majority of peripheral Vgamma9/Vdelta2 T cells shares migration properties with circulating monocytes, which explains the presence of these two distinct blood cell types in the inflammatory infiltrate at sites of infection and suggests that they synergize in anti-microbial immune responses. Our present findings demonstrate a rapid and HMB-PP-dependent crosstalk between Vgamma9/Vdelta2 T cells and autologous monocytes that results in the immediate production of inflammatory mediators including the cytokines interleukin (IL-6, interferon (IFN-gamma, tumor necrosis factor (TNF-alpha, and oncostatin M (OSM; the chemokines CCL2, CXCL8, and CXCL10; and TNF-related apoptosis-inducing ligand (TRAIL. Moreover, under these co-culture conditions monocytes differentiate within 18 hours into inflammatory dendritic cells (DCs with antigen-presenting functions. Addition of further microbial stimuli (lipopolysaccharide, peptidoglycan induces CCR7 and enables these inflammatory DCs to trigger the generation of CD4(+ effector alphabeta T cells expressing IFN-gamma and/or IL-17. Importantly, our in vitro model replicates the responsiveness to microbes of effluent cells from peritoneal dialysis (PD patients and translates directly to episodes of acute PD-associated bacterial peritonitis, where Vgamma9/Vdelta2 T cell numbers and soluble inflammatory mediators are elevated in patients infected with HMB-PP-producing pathogens. Collectively, these findings suggest a direct link between invading pathogens, microbe

  13. Geothermal Grows Up

    Science.gov (United States)

    Johnson, William C.; Kraemer, Steven; Ormond, Paul

    2011-01-01

    Self-declared energy and carbon reduction goals on the part of progressive colleges and universities have driven ground source geothermal space heating and cooling systems into rapid evolution, as part of long-term climate action planning efforts. The period of single-building or single-well solutions is quickly being eclipsed by highly engineered…

  14. Rapid assay for simultaneous detection and differentiation of immunoglobulin G antibodies to human immunodeficiency virus type 1 (HIV-1) group M, HIV-1 group O, and HIV-2.

    Science.gov (United States)

    Vallari, A S; Hickman, R K; Hackett, J R; Brennan, C A; Varitek, V A; Devare, S G

    1998-12-01

    A rapid immunodiagnostic test that detects and discriminates human immunodeficiency virus (HIV) infections on the basis of viral type, HIV type 1 (HIV-1) group M, HIV-1 group O, or HIV-2, was developed. The rapid assay for the detection of HIV (HIV rapid assay) was designed as an instrument-free chromatographic immunoassay that detects immunoglobulin G (IgG) antibodies to HIV. To assess the performance of the HIV rapid assay, 470 HIV-positive plasma samples were tested by PCR and/or Western blotting to confirm the genotype of the infecting virus. These samples were infected with strains that represented a wide variety of HIV strains including HIV-1 group M (subtypes A through G), HIV-1 group O, and HIV-2 (subtypes A and B). The results showed that the HIV genotype identity established by the rapid assay reliably (469 of 470 samples) correlates with the HIV genotype identity established by PCR or Western blotting. A total of 879 plasma samples were tested for IgG to HIV by a licensed enzyme immunoassay (EIA) (470 HIV-positive samples and 409 HIV-negative samples). When they were tested by the rapid assay, 469 samples were positive and 410 were negative (99.88% agreement). Twelve seroconversion panels were tested by both the rapid assay and a licensed EIA. For nine panels identical results were obtained by the two assays. For the remaining three panels, the rapid assay was positive one bleed later in comparison to the bleed at which the EIA was positive. One hundred three urine samples, including 93 urine samples from HIV-seropositive individuals and 10 urine samples from seronegative individuals, were tested by the rapid assay. Ninety-one of the ninety-three urine samples from HIV-seropositive individuals were found to be positive by the rapid assay. There were no false-positive results (98.05% agreement). Virus in all urine samples tested were typed as HIV-1 group M. These results suggest that a rapid assay based on the detection of IgG specific for selected

  15. The vulnerable, rapidly growing thoracic spine of the adolescent

    African Journals Online (AJOL)

    at fault, refusing to allow a 'trivial complaint' to stand in the way of the success of the team in which their sons play a key role. Department of Orthopaedics, University .... hence the diagnosis of 'postural, non-structural, thoracic kyphosis'. In a small percentage of these cases, structural changes have been seen later, but with ...

  16. The vulnerable, rapidly growing thoracic spine of the adolescent ...

    African Journals Online (AJOL)

    Prolapse of disc tissue occurs into the verfebral body, causing a disturbance of growth but little if any pain. The vertebrae in the mid-thoracic region become wedge-shaped, and a kyphotic deformity results, the so-called Scheuermann's disease, or adolescent kyphosis. A plea is made for the screening of children exposed to ...

  17. Effects of photodynamic therapy on rapidly growing nontuberculous mycobacteria keratitis.

    Science.gov (United States)

    Shih, Min-Hsiu; Huang, Fu-Chin

    2011-01-05

    The authors investigated the antimicrobial effect of methylene blue (MB)-mediated photodynamic therapy (PDT) on Mycobacterium fortuitum keratitis. In the in vitro study, the mycobacterial suspension and colonies were treated with the following: no MB, no light (normal control); MB and no light (dye control); light and no MB (light control); MB and light (PDT). Morphologic characteristics were examined by transmission electron microscopy. The bactericidal effects of combined PDT and antibiotic therapy (ciprofloxacin, moxifloxacin, and amikacin) were determined using the broth microdilution technique. Twenty-one rabbits with Mycobacterium keratitis were randomly divided into three groups (no treatment, topical amikacin treatment, and PDT combined with amikacin treatment). The clinical features of keratitis were scored and graded before treatment and before euthanatization. The diseased corneas were trephined for quantitative bacteriologic analysis to determine the antibacterial efficacy of the treatment. In the in vitro tests, the bacterial count had a 2-log reduction immediately after PDT treatment at 100 J/cm(2) with 10(-3)% MB. After PDT at 100 J/cm(2) with 10(-2)% MB, almost no viable bacteria were detected. PDT had a synergistic antimicrobial effect in combination with antibiotics. The phototoxicity occurred in the cytoplasm first and then disrupted the mycobacterial cell walls by lysis. In the rabbit keratitis model, combined PDT resulted in significantly less bacterial burden (P PDT against Mycobacterium fortuitum. PDT could be a potential alternative treatment for nontuberculous mycobacterial corneal infections.

  18. Targeted disruption in mice of a neural stem cell-maintaining, KRAB-Zn finger-encoding gene that has rapidly evolved in the human lineage.

    Directory of Open Access Journals (Sweden)

    Huan-Chieh Chien

    Full Text Available Understanding the genetic basis of the physical and behavioral traits that separate humans from other primates is a challenging but intriguing topic. The adaptive functions of the expansion and/or reduction in human brain size have long been explored. From a brain transcriptome project we have identified a KRAB-Zn finger protein-encoding gene (M003-A06 that has rapidly evolved since the human-chimpanzee separation. Quantitative RT-PCR analysis of different human tissues indicates that M003-A06 expression is enriched in the human fetal brain in addition to the fetal heart. Furthermore, analysis with use of immunofluorescence staining, neurosphere culturing and Western blotting indicates that the mouse ortholog of M003-A06, Zfp568, is expressed mainly in the embryonic stem (ES cells and fetal as well as adult neural stem cells (NSCs. Conditional gene knockout experiments in mice demonstrates that Zfp568 is both an NSC maintaining- and a brain size-regulating gene. Significantly, molecular genetic analyses show that human M003-A06 consists of 2 equilibrated allelic types, H and C, one of which (H is human-specific. Combined contemporary genotyping and database mining have revealed interesting genetic associations between the different genotypes of M003-A06 and the human head sizes. We propose that M003-A06 is likely one of the genes contributing to the uniqueness of the human brain in comparison to other higher primates.

  19. Growing unculturable bacteria.

    Science.gov (United States)

    Stewart, Eric J

    2012-08-01

    The bacteria that can be grown in the laboratory are only a small fraction of the total diversity that exists in nature. At all levels of bacterial phylogeny, uncultured clades that do not grow on standard media are playing critical roles in cycling carbon, nitrogen, and other elements, synthesizing novel natural products, and impacting the surrounding organisms and environment. While molecular techniques, such as metagenomic sequencing, can provide some information independent of our ability to culture these organisms, it is essentially impossible to learn new gene and pathway functions from pure sequence data. A true understanding of the physiology of these bacteria and their roles in ecology, host health, and natural product production requires their cultivation in the laboratory. Recent advances in growing these species include coculture with other bacteria, recreating the environment in the laboratory, and combining these approaches with microcultivation technology to increase throughput and access rare species. These studies are unraveling the molecular mechanisms of unculturability and are identifying growth factors that promote the growth of previously unculturable organisms. This minireview summarizes the recent discoveries in this area and discusses the potential future of the field.

  20. Aptamer based assay of plated-derived grow factor in unprocessed human plasma sample and MCF-7 breast cancer cell lysates using gold nanoparticle supported α-cyclodextrin.

    Science.gov (United States)

    Hasanzadeh, Mohammad; Razmi, Nasrin; Mokhtarzadeh, Ahad; Shadjou, Nasrin; Mahboob, Soltanali

    2017-11-24

    unprocessed human plasma samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Problems of rapid growth.

    Science.gov (United States)

    Kim, T D

    1980-01-01

    South Korea's export-oriented development strategy has achieved a remarkable growth record, but it has also brought 2 different problems: 1) since the country's exports accounted for about 1% of total world export volume, the 1st world has become fearful about Korea's aggressive export drive; and 2) the fact that exports account for over 30% of its total gross national product (GNP) exposes the vulnerability of South Korea's economy itself. South Korea continues to be a poor nation, although it is rated as 1 of the most rapidly growing middle income economies. A World Bank 1978 report shows Korea to be 28th of 58 middle income countries in terms of per capita GNP in 1976. Of 11 newly industrializing countries (NIC), 5 in the European continent are more advanced than the others. A recent emphasis on the basic human needs approach has tended to downgrade the concept of GNP. Korea has only an abundant labor force and is without any natural resources. Consequently, Korea utilized an export-oriented development strategy. Oil requirements are met with imports, and almost all raw materials to be processed into exportable products must be imported. To pay import bills Korea must export and earn foreign exchange. It must be emphasized that foreign trade must always be 2-way traffic. In order to export more to middle income countries like Korea, the countries of the 1st world need to ease their protectionist measures against imports from developing countries.

  2. The SGIA and the Common Growing Language

    Directory of Open Access Journals (Sweden)

    Laurentiu CIOVICA

    2011-01-01

    Full Text Available Human or virtual agents are presented in our lives daily. They serve our purposes and represent us in different many situations. Nowadays the number of virtual agents is increasing daily because they are cheaper, faster and more accurate than human agents. Our aim in this article is to define a new type of intelligent agent called SGIA – Self Growing Intelligent Agent and a new defining language for it. The SGIA agent is an intelligent agent with all the common agents’ characteristics and with other special one: that to learn and grow by itself in knowledge and size.

  3. Analysis of 62 hybrid assembled human Y chromosomes exposes rapid structural changes and high rates of gene conversion

    DEFF Research Database (Denmark)

    Gonzalez-Izarzugaza, Jose Maria; Skov, Laurits; Maretty, Lasse

    2017-01-01

    The human Y-chromosome does not recombine across its male-specific part and is therefore an excellent marker of human migrations. It also plays an important role in male fertility. However, its evolution is difficult to fully understand because of repetitive sequences, inverted repeats...

  4. Rapid milk group classification by 1H NMR analysis of Le and H epitopes in human milk oligosaccharide donor samples

    NARCIS (Netherlands)

    van Leeuwen, Sander S; Schoemaker, Ruud J W; Gerwig, Gerrit J; van Leusen-van Kan, Ellen J M; Dijkhuizen, Lubbert; Kamerling, Johannis P

    Human milk oligosaccharides (HMOs) are a major constituent of human breast milk and play an important role in reducing the risk of infections in infants. The structures of these HMOs show similarities with blood group antigens in protein glycosylation, in particular in relation to fucosylation in

  5. Design and implementation of a controlled clinical trial to evaluate the effectiveness and efficiency of routine opt-out rapid human immunodeficiency virus screening in the emergency department.

    Science.gov (United States)

    Haukoos, Jason S; Hopkins, Emily; Byyny, Richard L; Conroy, Amy A; Silverman, Morgan; Eisert, Sheri; Thrun, Mark; Wilson, Michael; Boyett, Brian; Heffelfinger, James D

    2009-08-01

    In 2006, the Centers for Disease Control and Prevention (CDC) released revised recommendations for performing human immunodeficiency virus (HIV) testing in health care settings, including implementing routine rapid HIV screening, the use of an integrated opt-out consent, and limited prevention counseling. Emergency departments (EDs) have been a primary focus of these efforts. These revised CDC recommendations were primarily based on feasibility studies and have not been evaluated through the application of rigorous research methods. This article describes the design and implementation of a large prospective controlled clinical trial to evaluate the CDC's recommendations in an ED setting. From April 15, 2007, through April 15, 2009, a prospective quasi-experimental equivalent time-samples clinical trial was performed to compare the clinical effectiveness and efficiency of routine (nontargeted) opt-out rapid HIV screening (intervention) to physician-directed diagnostic rapid HIV testing (control) in a high-volume urban ED. In addition, three nested observational studies were performed to evaluate the cost-effectiveness and patient and staff acceptance of the two rapid HIV testing methods. This article describes the rationale, methodologies, and study design features of this program evaluation clinical trial. It also provides details regarding the integration of the principal clinical trial and its nested observational studies. Such ED-based trials are rare, but serve to provide valid comparisons between testing approaches. Investigators should consider similar methodology when performing future ED-based health services research.

  6. Rapid localized spread and immunologic containment define Herpes simplex virus-2 reactivation in the human genital tract.

    Science.gov (United States)

    Schiffer, Joshua T; Swan, David; Al Sallaq, Ramzi; Magaret, Amalia; Johnston, Christine; Mark, Karen E; Selke, Stacy; Ocbamichael, Negusse; Kuntz, Steve; Zhu, Jia; Robinson, Barry; Huang, Meei-Li; Jerome, Keith R; Wald, Anna; Corey, Lawrence

    2013-04-16

    Herpes simplex virus-2 (HSV-2) is shed episodically, leading to occasional genital ulcers and efficient transmission. The biology explaining highly variable shedding patterns, in an infected person over time, is poorly understood. We sampled the genital tract for HSV DNA at several time intervals and concurrently at multiple sites, and derived a spatial mathematical model to characterize dynamics of HSV-2 reactivation. The model reproduced heterogeneity in shedding episode duration and viral production, and predicted rapid early viral expansion, rapid late decay, and wide spatial dispersion of HSV replication during episodes. In simulations, HSV-2 spread locally within single ulcers to thousands of epithelial cells in genital epithelium. DOI:http://dx.doi.org/10.7554/eLife.00288.001.

  7. Improved method for rapid and accurate isolation and identification of Streptococcus mutans and Streptococcus sobrinus from human plaque samples.

    Science.gov (United States)

    Villhauer, Alissa L; Lynch, David J; Drake, David R

    2017-08-01

    Mutans streptococci (MS), specifically Streptococcus mutans (SM) and Streptococcus sobrinus (SS), are bacterial species frequently targeted for investigation due to their role in the etiology of dental caries. Differentiation of S. mutans and S. sobrinus is an essential part of exploring the role of these organisms in disease progression and the impact of the presence of either/both on a subject's caries experience. Of vital importance to the study of these organisms is an identification protocol that allows us to distinguish between the two species in an easy, accurate, and timely manner. While conducting a 5-year birth cohort study in a Northern Plains American Indian tribe, the need for a more rapid procedure for isolating and identifying high volumes of MS was recognized. We report here on the development of an accurate and rapid method for MS identification. Accuracy, ease of use, and material and time requirements for morphological differentiation on selective agar, biochemical tests, and various combinations of PCR primers were compared. The final protocol included preliminary identification based on colony morphology followed by PCR confirmation of species identification using primers targeting regions of the glucosyltransferase (gtf) genes of SM and SS. This method of isolation and identification was found to be highly accurate, more rapid than the previous methodology used, and easily learned. It resulted in more efficient use of both time and material resources. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Meeting a Growing Need

    National Research Council Canada - National Science Library

    Geyer, Sally C

    2007-01-01

    ... to consider are: * Are they SAS70 certified? * Are they highly knowledgeable in the outsourced area and its regulations? * Do they have an infrastructure that will support growth? * Are they technologically advanced enough to automate as much of the process as possible, in order to avoid human error? * Can they offer the client visibility ...

  9. Growing for different ends.

    Science.gov (United States)

    Catts, Oron; Zurr, Ionat

    2014-11-01

    Tissue engineering and regenerative biology are usually discussed in relation to biomedical research and applications. However, hand in hand with developments of this field in the biomedical context, other approaches and uses for non-medical ends have been explored. There is a growing interest in exploring spin off tissue engineering and regenerative biology technologies in areas such as consumer products, art and design. This paper outlines developments regarding in vitro meat and leather, actuators and bio-mechanic interfaces, speculative design and contemporary artistic practices. The authors draw on their extensive experience of using tissue engineering for non-medical ends to speculate about what lead to these applications and their possible future development and uses. Avoiding utopian and dystopian postures and using the notion of the contestable, this paper also mentions some philosophical and ethical consideration stemming from the use of non-medical approaches to tissue constructs. This article is part of a directed issue entitled: Regenerative Medicine: the challenge of translation. Copyright © 2014. Published by Elsevier Ltd.

  10. Recent developments in the study of rapid human movements with the kinematic theory: Applications to handwriting and signature synthesis

    OpenAIRE

    Plamondon, Réjean; O’Reilly, Christian; Galbally, Javier; Almaksour, Abdullah; Anquetil, Éric

    2014-01-01

    International audience; Human movement modeling can be of great interest for the design of pattern recognition systems relying on the understanding of the fine motor control (such as on-line handwriting recognition or signature verification) as well as for the development of intelligent systems involving in a way or another the processing of human movements. In this paper, we briefly list the different models that have been proposed in order to characterize the handwriting process and focus o...

  11. The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage

    DEFF Research Database (Denmark)

    Christiansen, Stig Hill; Zhang, Xianwei; Juul-Madsen, Kristian

    2017-01-01

    in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell...... of certain oligomeric and chemical properties to support cytotoxic effects of cationic polymers targeting human leukocytes....

  12. Controlling human corneal stromal stem cell contraction to mediate rapid cell and matrix organization of real architecture for 3-dimensional tissue equivalents.

    Science.gov (United States)

    Mukhey, Dev; Phillips, James B; Daniels, Julie T; Kureshi, Alvena K

    2017-12-05

    The architecture of the human corneal stroma consists of a highly organized extracellular matrix (ECM) interspersed with keratocytes. Their progenitor cells; corneal stromal stem cells (CSSC) are located at the periphery, in the limbal stroma. A highly organized corneal ECM is critical for effective transmission of light but this structure may be compromised during injury or disease, resulting in loss of vision. Re-creating normal organization in engineered tissue equivalents for transplantation often involves lengthy culture times that are inappropriate for clinical use or utilisation of synthetic substrates that bring complications such as corneal melting. CSSC have great therapeutic potential owing to their ability to reorganize a disorganized matrix, restoring transparency in scarred corneas. We examined CSSC contractile behavior to assess whether this property could be exploited to rapidly generate cell and ECM organization in Real Architecture For 3D Tissues (RAFT) tissue equivalents (TE) for transplantation. Free-floating collagen gels were characterized to assess contractile behavior of CSSC and establish optimum cell density and culture times. To mediate cell and collagen organization, tethered collagen gels seeded with CSSC were cultured and subsequently stabilized with the RAFT process. We demonstrated rapid creation of biomimetic RAFT TE with tunable structural properties. These displayed three distinct regions of varying degrees of cellular and collagen organization. Interestingly, increased organization coincided with a dramatic loss of PAX6 expression in CSSC, indicating rapid differentiation into keratocytes. The organized RAFT TE system could be a useful bioengineering tool to rapidly create an organized ECM while simultaneously controlling cell phenotype. For the first time, we have demonstrated that human CSSC exhibit the phenomenon of cellular self-alignment in tethered collagen gels. We found this mediated rapid co-alignment of collagen fibrils

  13. Case grows for climate change

    Energy Technology Data Exchange (ETDEWEB)

    Hileman, B.

    1999-08-09

    In the four years since the IPCC stated that 'the balance of evidence suggests a discernible human influence on global climate', evidence for anomalous warming has become more compelling, and as a result scientists have become more concerned that human-induced climate change has already arrived. The article summarises recent extra evidence on global temperatures, carbon dioxide measurements, ice shelf breakup, coral bleaching, unstable climates and improved climate models. At the time of the Kyoto conference, the US became keen on the idea that enhancing forest and soil carbon sequestration was a good way to offset emissions reduction targets. Congress is however under the opinion on that the Kyoto protocol presents a threat to the US economy, and senate is very unlikely to ratify the protocol during the Clinton Administration. The debate as to whether the US government should mandate major emission reduction or wait for more scientific certainty may continue for a number of years, but, growing concern of scientists and the public for the harmful effects of climate change may cause a change. 4 figs., 8 photos.

  14. Cerebral O2 metabolism and cerebral blood flow in humans during deep and rapid-eye-movement sleep

    DEFF Research Database (Denmark)

    Madsen, P L; Schmidt, J F; Wildschiødtz, Gordon

    1991-01-01

    It could be expected that the various stages of sleep were reflected in variation of the overall level of cerebral activity and thereby in the magnitude of cerebral metabolic rate of oxygen (CMRO2) and cerebral blood flow (CBF). The elusive nature of sleep imposes major methodological restrictions...... on examination of this question. We have now measured CBF and CMRO2 in young healthy volunteers using the Kety-Schmidt technique with 133Xe as the inert gas. Measurements were performed during wakefulness, deep sleep (stage 3/4), and rapid-eye-movement (REM) sleep as verified by standard polysomnography...

  15. BIOTECHNOLOGY IN FRUIT GROWING

    Directory of Open Access Journals (Sweden)

    Z. Jurković

    2008-09-01

    Full Text Available Research studies in the area of biotechnologies in fruit growing started at the Agricultural Institute Osijek in 2006 with the establishment of the first experimental in vitro laboratory for micropropagation. The laboratory started an active research related to the Project "Biotechnological methods in fruit tree identification, selection and propagation" Project is part of program "Preservation and revitalization of grape and fruit autochthonous cultivars". The goal of this research is to determine genetic differences between autochthonous and introduced cultivars of cherry as well as cultivars and types of sour cherry, to find and optimize a method for fast recovery of clonal material. A great number of cherry cultivars and types within the population of cv. Oblacinska sour cherry exists in Croatia. A survey with the purpose of selecting autochthonous cultivars for further selection has been done in previous research. Differences have been found in a number of important agronomic traits within the populations of cv. Oblačinska sour cherry. Autochthonous cherry cultivars are suspected to be synonyms of known old cultivars which were introduced randomly and have been naturalized under a local name. Identification and description of cultivars and types of fruits is based on special visible properties which were measurable or notable. In this approach difficulties arise from the effect of non-genetic factors on expression of certain traits. Genetic-physiological problem of S allele autoincompatibility exists within cherry cultivars. Therefore it is necessary to put different cultivars in the plantation to pollinate each other. Apart form the fast and certain sort identification independent of environmental factors, biotechnological methods based on PCR enable faster virus detection compared with classical serologic methods and indexing and cover a wider range of plant pathogens including those undetectable by other methods. Thermotherapy and

  16. Impact assessment of human diet changes with rapid urbanization on regional nitrogen and phosphorus flows--a case study of the megacity Shanghai.

    Science.gov (United States)

    Liu, Chen; Zou, Chunjing; Wang, Qinxue; Hayashi, Yoshitsugu; Yasunari, Tetsuzo

    2014-02-01

    Regional material flows are strongly influenced by human diets. To diagnose and prevent environmental problems that threaten urban sustainability, the impact of human diet changes with rapid urbanization on the regional nitrogen (N) and phosphorus (P) flows were quantitatively evaluated. A survey of day-to-day activities was conducted of 450 individuals surveyed (adults over 18 years old) in three representative areas (the central district, the new district, and the suburban/rural areas) of Shanghai, a megacity which has attracted worldwide attention. The lifestyle (eating habits, domestic sanitation, drainage facilities, etc.) pattern was determined and the potential N and P loads from human diets on the environment were calculated. The daily potential nitrogen and phosphorus loads from human diets was 19.36 g-N, 1.80 g-P in the central district, 16.48 g-N, 1.52 g-P in the new district, and 13.04 g-N, 1.20 g-P in the suburban/rural areas of Shanghai. Respondents in all three areas, especially those in the suburban/rural areas reported a preference for increasing the intake of animal-derived as well as processed foods, which means that the potential N and P load from human diets to the environment will increase further. In addition, most respondents consider industrial wastewater discharge as the main cause of eutrophication of waterbodies, though in recent years water pollution caused by domestic wastewater has increased rapidly, but this has received much less attention. Environment-friendly eating habits and improvements in the environmental awareness will be required.

  17. Growing Galaxies Gently

    Science.gov (United States)

    2010-10-01

    New observations from ESO's Very Large Telescope have, for the first time, provided direct evidence that young galaxies can grow by sucking in the cool gas around them and using it as fuel for the formation of many new stars. In the first few billion years after the Big Bang the mass of a typical galaxy increased dramatically and understanding why this happened is one of the hottest problems in modern astrophysics. The results appear in the 14 October issue of the journal Nature. The first galaxies formed well before the Universe was one billion years old and were much smaller than the giant systems - including the Milky Way - that we see today. So somehow the average galaxy size has increased as the Universe has evolved. Galaxies often collide and then merge to form larger systems and this process is certainly an important growth mechanism. However, an additional, gentler way has been proposed. A European team of astronomers has used ESO's Very Large Telescope to test this very different idea - that young galaxies can also grow by sucking in cool streams of the hydrogen and helium gas that filled the early Universe and forming new stars from this primitive material. Just as a commercial company can expand either by merging with other companies, or by hiring more staff, young galaxies could perhaps also grow in two different ways - by merging with other galaxies or by accreting material. The team leader, Giovanni Cresci (Osservatorio Astrofisico di Arcetri) says: "The new results from the VLT are the first direct evidence that the accretion of pristine gas really happened and was enough to fuel vigorous star formation and the growth of massive galaxies in the young Universe." The discovery will have a major impact on our understanding of the evolution of the Universe from the Big Bang to the present day. Theories of galaxy formation and evolution may have to be re-written. The group began by selecting three very distant galaxies to see if they could find evidence

  18. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow

    Science.gov (United States)

    Colter, David C.; Class, Reiner; DiGirolamo, Carla M.; Prockop, Darwin J.

    2000-01-01

    Cultures of plastic-adherent cells from bone marrow have attracted interest because of their ability to support growth of hematopoietic stem cells, their multipotentiality for differentiation, and their possible use for cell and gene therapy. Here we found that the cells grew most rapidly when they were initially plated at low densities (1.5 or 3.0 cells/cm2) to generate single-cell derived colonies. The cultures displayed a lag phase of about 5 days, a log phase of rapid growth of about 5 days, and then a stationary phase. FACS analysis demonstrated that stationary cultures contained a major population of large and moderately granular cells and a minor population of small and agranular cells here referred to as recycling stem cells or RS-1 cells. During the lag phase, the RS-1 cells gave rise to a new population of small and densely granular cells (RS-2 cells). During the late log phase, the RS-2 cells decreased in number and regenerated the pool of RS-1 cells found in stationary cultures. In repeated passages in which the cells were plated at low density, they were amplified about 109-fold in 6 wk. The cells retained their ability to generate single-cell derived colonies and therefore apparently retained their multipotentiality for differentiation. PMID:10725391

  19. Rapid high-resolution three-dimensional mapping of T1 and age-dependent variations in the non-human primate brain using magnetization-prepared rapid gradient-echo (MPRAGE) sequence

    Science.gov (United States)

    Liu, Junjie V.; Bock, Nicholas A.; Silva, Afonso C.

    2011-01-01

    The use of quantitative T1 mapping in neuroscience and neurology has raised strong interest in the development of T1-mapping techniques that can measure T1 in the whole brain, with high accuracy and precision and within short imaging and computation times. Here, we present a new inversion-recovery (IR) based T1-mapping method using a standard 3D magnetization-prepared rapid gradient-echo (MPRAGE) sequence. By varying only the inversion time (TI), but keeping other parameters constant, MPRAGE image signals become linear to exp(−TI/T1), allowing for accurate T1 estimation without flip angle correction. We also show that acquiring data at just 3 TIs, with the three different TI values optimized, gives maximum T1 precision per unit time, allowing for new efficient approaches to measure and compute T1. We demonstrate the use of our method at 7 Tesla to obtain 3D T1 maps of the whole brain in common marmosets at 0.60 mm resolution and within 11 minutes. T1 maps from the same individuals were highly reproducible across different days. Across subjects, the peak of cerebral gray matter T1 distribution was 1735±52 ms, and the lower edge of cerebral white matter T1 distribution was 1270±43 ms. We found a significant decrease of T1 in both gray and white matter of the marmoset brain with age over a span of 14 years, in agreement with previous human studies. This application illustrates that MPRAGE-based 3D T1 mapping is rapid, accurate and precise, and can facilitate high-resolution anatomical studies in neuroscience and neurological diseases. PMID:21376814

  20. Rapid Industrial Development, Competition, and Relative Economic Status: A Study in Human Ecology. Working Paper RID 73.10.

    Science.gov (United States)

    Summers, Gene F.; Clemente, Frank

    The document has 3 goals (1) to examine the distribution of economic benefits of industrial development as reflected by the concept of competition in human ecology; (2) to provide an empirical test of the ecological model, and (3) to relate the findings to public policy. Two Illinois study areas were identified. First, as an experimental region,…

  1. The cost effectiveness of rapid-acting insulin aspart compared with human insulin in type 2 diabetes patients: an analysis from the Japanese third-party payer perspective.

    Science.gov (United States)

    Pollock, R F; Valentine, W J; Pilgaard, T; Nishimura, H

    2011-01-01

    The Nippon Ultra-Rapid Insulin and Diabetic Complication Evaluation Study (NICE Study) (NCT00575172) was a 5-year, open-label, randomised controlled trial which compared cardiovascular outcomes in Japanese type 2 diabetes patients intensively treated with regular human insulin or insulin aspart (NovoRapid; Novo Nordisk A/S, Bagsvaerd, Denmark), a rapid-acting insulin analogue. The aim of the present analysis was to evaluate the cost effectiveness of insulin aspart versus regular human insulin from the perspective of a Japanese third-party healthcare payer. A discrete event-simulation model was developed in Microsoft Excel to assess the within-trial cost effectiveness and make longer-term clinical projections in patients treated with regular human insulin or insulin aspart. In addition to severe hypoglycaemia, the model captured myocardial and cerebral infarction events and percutaneous coronary intervention and coronary artery bypass graft procedures. Within-trial mortality, incidence of severe hypoglycaemia and cardiovascular event probabilities were derived from the annual rates observed during the trial period, while post-trial outcomes were calculated using the event rates from the trial, adjusted for increasing patient age. Event costs were accounted from the healthcare payer perspective and expressed in 2008 Japanese yen (JPY), while health-related quality of life (HRQoL) was captured using event and state utilities. Future costs and clinical benefits were discounted at 3% annually. Life expectancy, quality-adjusted life expectancy, cardiovascular event rates and costs were evaluated over 5- and 10-year time horizons and sensitivity analyses were performed to assess variability in model outcomes. Over 5 years of treatment, insulin aspart dominated human insulin both in incremental life expectancy and in incremental quality-adjusted life-years (QALYS). Insulin aspart was associated with a small improvement in discounted life expectancy of 0.005 years (4.688 vs

  2. Critical role of perforin-dependent CD8+ T cell immunity for rapid protective vaccination in a murine model for human smallpox.

    Directory of Open Access Journals (Sweden)

    Melanie Kremer

    Full Text Available Vaccination is highly effective in preventing various infectious diseases, whereas the constant threat of new emerging pathogens necessitates the development of innovative vaccination principles that also confer rapid protection in a case of emergency. Although increasing evidence points to T cell immunity playing a critical role in vaccination against viral diseases, vaccine efficacy is mostly associated with the induction of antibody responses. Here we analyze the immunological mechanism(s of rapidly protective vaccinia virus immunization using mousepox as surrogate model for human smallpox. We found that fast protection against lethal systemic poxvirus disease solely depended on CD4 and CD8 T cell responses induced by vaccination with highly attenuated modified vaccinia virus Ankara (MVA or conventional vaccinia virus. Of note, CD4 T cells were critically required to allow for MVA induced CD8 T cell expansion and perforin-mediated cytotoxicity was a key mechanism of MVA induced protection. In contrast, selected components of the innate immune system and B cell-mediated responses were fully dispensable for prevention of fatal disease by immunization given two days before challenge. In conclusion, our data clearly demonstrate that perforin-dependent CD8 T cell immunity plays a key role in MVA conferred short term protection against lethal mousepox. Rapid induction of T cell immunity might serve as a new paradigm for treatments that need to fit into a scenario of protective emergency vaccination.

  3. Critical role of perforin-dependent CD8+ T cell immunity for rapid protective vaccination in a murine model for human smallpox.

    Science.gov (United States)

    Kremer, Melanie; Suezer, Yasemin; Volz, Asisa; Frenz, Theresa; Majzoub, Monir; Hanschmann, Kay-Martin; Lehmann, Michael H; Kalinke, Ulrich; Sutter, Gerd

    2012-01-01

    Vaccination is highly effective in preventing various infectious diseases, whereas the constant threat of new emerging pathogens necessitates the development of innovative vaccination principles that also confer rapid protection in a case of emergency. Although increasing evidence points to T cell immunity playing a critical role in vaccination against viral diseases, vaccine efficacy is mostly associated with the induction of antibody responses. Here we analyze the immunological mechanism(s) of rapidly protective vaccinia virus immunization using mousepox as surrogate model for human smallpox. We found that fast protection against lethal systemic poxvirus disease solely depended on CD4 and CD8 T cell responses induced by vaccination with highly attenuated modified vaccinia virus Ankara (MVA) or conventional vaccinia virus. Of note, CD4 T cells were critically required to allow for MVA induced CD8 T cell expansion and perforin-mediated cytotoxicity was a key mechanism of MVA induced protection. In contrast, selected components of the innate immune system and B cell-mediated responses were fully dispensable for prevention of fatal disease by immunization given two days before challenge. In conclusion, our data clearly demonstrate that perforin-dependent CD8 T cell immunity plays a key role in MVA conferred short term protection against lethal mousepox. Rapid induction of T cell immunity might serve as a new paradigm for treatments that need to fit into a scenario of protective emergency vaccination.

  4. Rapid quantitative PCR assays for the simultaneous detection of herpes simplex virus, varicella zoster virus, cytomegalovirus, Epstein-Barr virus, and human herpesvirus 6 DNA in blood and other clinical specimens

    NARCIS (Netherlands)

    Engelmann, I.; Petzold, D. R.; Kosinska, A.; Hepkema, B. G.; Schulz, T. F.; Heim, A.

    Rapid diagnosis of human herpesvirus primary infections or reactivations is facilitated by quantitative PCRs. Quantitative PCR assays with a standard thermal cycling profile permitting simultaneous detection of herpes simplex virus (HSV), varicella zoster virus (VZV), cytomegalovirus (CMV),

  5. Rapid and sensitive determination of levofloxacin in microsamples of human plasma by high-performance liquid chromatography and its application in a pharmacokinetic study.

    Science.gov (United States)

    Aguilar-Carrasco, José Carlos; Hernández-Pineda, Jessica; Jiménez-Andrade, Juan Miguel; Flores-Murrieta, Francisco Javier; Carrasco-Portugal, Miriam Del Carmen; López-Canales, Jorge Skiold

    2015-03-01

    A rapid, sensitive and simple high-performance liquid chromatographic assay with ultraviolet detection was developed for the quantification of levofloxacin in microsamples (100 μL) of human plasma. The extraction procedure included a protein precipitation technique and a short chromatographic running time (4.5 min). Analyses were carried out on a Symmetry C18 column using a mixture of acetonitrile and 0.01 m potassium dihydrogen aqueous solution (pH 3.4; 14:86 v/v) as mobile phase. The method provided specificity and was linear (r ≥ 0.9992) over the concentration range 0.1-12 µg/mL. The average absolute recovery was 93.59%. The intra- and inter-day coefficients of variation were levofloxacin was stable in all evaluations. The usefulness of this method was demonstrated in a pharmacokinetic study of levofloxacin in healthy adult volunteers. The present method offers two main advantages: (a) the use of microsamples reduces the total volume of blood to be collected from patients; and (b) it provides a good cost-effectiveness ratio. It is concluded that the method is rapid, simple, sensitive, economical and suitable for the determination of levofloxacin in human plasma using a small volume of sample. Copyright © 2014 John Wiley & Sons, Ltd.

  6. Rapid Cellular Phenotyping of Human Pluripotent Stem Cell-Derived Cardiomyocytes using a Genetically Encoded Fluorescent Voltage Sensor

    Directory of Open Access Journals (Sweden)

    Jordan S. Leyton-Mange

    2014-02-01

    Full Text Available In addition to their promise in regenerative medicine, pluripotent stem cells have proved to be faithful models of many human diseases. In particular, patient-specific stem cell-derived cardiomyocytes recapitulate key features of several life-threatening cardiac arrhythmia syndromes. For both modeling and regenerative approaches, phenotyping of stem cell-derived tissues is critical. Cellular phenotyping has largely relied upon expression of lineage markers rather than physiologic attributes. This is especially true for cardiomyocytes, in part because electrophysiological recordings are labor intensive. Likewise, most optical voltage indicators suffer from phototoxicity, which damages cells and degrades signal quality. Here we present the use of a genetically encoded fluorescent voltage indicator, ArcLight, which we demonstrate can faithfully report transmembrane potentials in human stem cell-derived cardiomyocytes. We demonstrate the application of this fluorescent sensor in high-throughput, serial phenotyping of differentiating cardiomyocyte populations and in screening for drug-induced cardiotoxicity.

  7. Cost-effectiveness analysis of cervical cancer prevention based on a rapid human papillomavirus screening test in a high-risk region of China.

    Science.gov (United States)

    Levin, Carol E; Sellors, John; Shi, Ju-Fang; Ma, Li; Qiao, You-lin; Ortendahl, Jesse; O'Shea, Meredith K H; Goldie, Sue J

    2010-09-01

    This study assessed the cost-effectiveness of a new, rapid human papillomavirus (HPV)-DNA screening test for cervical cancer prevention in the high-risk region of Shanxi, China. Using micro-costing methods, we estimated the resources needed to implement preventive strategies using cervical cytology or HPV-DNA testing, including the Hybrid Capture 2 (hc2) test (QIAGEN Corp., Gaithersburg, MD) and the rapid HPV-DNA careHPV test (QIAGEN). Data were used in a previously published model and empirically calibrated to country-specific epidemiological data. Strategies differed by initial test, targeted age, frequency of screening, number of clinic visits required (1, 2 or 3) and service delivery setting (national, county and township levels). Outcomes included lifetime risk of cancer, years of life saved (YLS), lifetime costs and incremental cost-effectiveness ratios (cost per YLS). For all screening frequencies, the most efficient strategy used 2-visit rapid HPV-DNA testing at the county level, including screening and diagnostics in the first visit, and treatment in the second visit. Screening at ages 35, 40 and 45 reduced cancer risk by 50% among women compliant with all 3 screening rounds, and was US$ 150 per YLS, compared with this same strategy applied twice per lifetime. This would be considered very cost-effective evaluated against China's per-capita gross domestic product (US$ 1,702). By enhancing the linkage between screening and treatment through a reduced number of visits, rapid HPV-DNA testing 3 times per lifetime is more effective than traditional cytology, and is likely to be cost-effective in high-risk regions of China.

  8. Magnetic solid phase extraction coupled with desorption corona beam ionization-mass spectrometry for rapid analysis of antidepressants in human body fluids.

    Science.gov (United States)

    Chen, Di; Zheng, Hao-Bo; Huang, Yun-Qing; Hu, Yu-Ning; Yu, Qiong-Wei; Yuan, Bi-Feng; Feng, Yu-Qi

    2015-08-21

    Ambient ionization techniques show good potential in rapid analysis of target compounds. However, a direct application of these ambient ionization techniques for the determination of analytes in a complex matrix is difficult due to the matrix interference and ion suppression. To resolve this problem, here we developed a strategy by coupling magnetic solid phase extraction (MSPE) with desorption corona beam ionization (DCBI)-mass spectrometry (MS). As a proof of concept, the pyrrole-coated Fe3O4 magnetic nanoparticles (Fe3O4@Ppy) were prepared and used for the extraction of antidepressants. After extraction, the Fe3O4@Ppy with trapped antidepressants was then directly subjected to DCBI-MS analysis with the aid of a homemade magnetic glass capillary. As the MSPE process is rapid and the direct DCBI-MS analysis does not need solvent desorption or chromatographic separation processes, the overall analysis can be completed within 3 min. The proposed MSPE-DCBI-MS method was then successfully used to determine antidepressants in human urine and plasma. The calibration curves were obtained in the range of 0.005-0.5 μg mL(-1) for urine and 0.02-1 μg mL(-1) for plasma with reasonable linearity (R(2) > 0.951). The limits of detection of three antidepressants were in the range of 0.2-1 ng mL(-1) for urine and 2-5 ng mL(-1) for plasma. Acceptable reproducibility for rapid analysis was achieved with relative standard deviations less than 19.1% and the relative recoveries were 85.2-118.7%. Taken together, the developed MSPE-DCBI-MS strategy offers a powerful capacity for rapid analysis of target compounds in a complex matrix, which would greatly expand the applications of ambient ionization techniques with plentiful magnetic sorbents.

  9. Rapid generation of mature hepatocyte-like cells from human induced pluripotent stem cells by an efficient three-step protocol.

    Science.gov (United States)

    Chen, Yu-Fan; Tseng, Chien-Yu; Wang, Hsei-Wei; Kuo, Hung-Chih; Yang, Vincent W; Lee, Oscar K

    2012-04-01

    Liver transplantation is the only definitive treatment for end-stage cirrhosis and fulminant liver failure, but the lack of available donor livers is a major obstacle to liver transplantation. Recently, induced pluripotent stem cells (iPSCs) derived from the reprogramming of somatic fibroblasts, have been shown to resemble embryonic stem (ES) cells in that they have pluripotent properties and the potential to differentiate into all cell lineages in vitro, including hepatocytes. Thus, iPSCs could serve as a favorable cell source for a wide range of applications, including drug toxicity testing, cell transplantation, and patient-specific disease modeling. Here, we describe an efficient and rapid three-step protocol that is able to rapidly generate hepatocyte-like cells from human iPSCs. This occurs because the endodermal induction step allows for more efficient and definitive endoderm cell formation. We show that hepatocyte growth factor (HGF), which synergizes with activin A and Wnt3a, elevates the expression of the endodermal marker Foxa2 (forkhead box a2) by 39.3% compared to when HGF is absent (14.2%) during the endodermal induction step. In addition, iPSC-derived hepatocytes had a similar gene expression profile to mature hepatocytes. Importantly, the hepatocyte-like cells exhibited cytochrome P450 3A4 (CYP3A4) enzyme activity, secreted urea, uptake of low-density lipoprotein (LDL), and possessed the ability to store glycogen. Moreover, the hepatocyte-like cells rescued lethal fulminant hepatic failure in a nonobese diabetic severe combined immunodeficient mouse model. We have established a rapid and efficient differentiation protocol that is able to generate functional hepatocyte-like cells from human iPSCs. This may offer an alternative option for treatment of liver diseases. Copyright © 2011 American Association for the Study of Liver Diseases.

  10. Rapid analysis of the interactions between drugs and human serum albumin (HSA) using high-performance affinity chromatography (HPAC)

    OpenAIRE

    Kim, Hee Seung; Wainer, Irving W.

    2008-01-01

    This study used a combination of zonal elution and frontal affinity chromatography on immobilized human serum albumin (HSA) high-performance affinity chromatography (HPAC) column to examine the association constants of various compounds that have been studied by equilibrium dialysis or ultra filtration. A standard plot was generated from retention factors of reference compounds using zonal elution chromatography against association constants of reference compounds using frontal affinity chrom...

  11. Rapid measurement of human milk macronutrients in the neonatal intensive care unit: accuracy and precision of fourier transform mid-infrared spectroscopy.

    Science.gov (United States)

    Smilowitz, Jennifer T; Gho, Deborah S; Mirmiran, Majid; German, J Bruce; Underwood, Mark A

    2014-05-01

    Although it is well established that human milk varies widely in macronutrient content, it remains common for human milk fortification for premature infants to be based on historic mean values. As a result, those caring for premature infants often underestimate protein intake. Rapid precise measurement of human milk protein, fat, and lactose to allow individualized fortification has been proposed for decades but remains elusive due to technical challenges. This study aimed to evaluate the accuracy and precision of a Fourier transform (FT) mid-infrared (IR) spectroscope in the neonatal intensive care unit to measure human milk fat, total protein, lactose, and calculated energy compared with standard chemical analyses. One hundred sixteen breast milk samples across lactation stages from women who delivered at term (n = 69) and preterm (n = 5) were analyzed with the FT mid-IR spectroscope and with standard chemical methods. Ten of the samples were tested in replicate using the FT mid-IR spectroscope to determine repeatability. The agreement between the FT mid-IR spectroscope analysis and reference methods was high for protein and fat and moderate for lactose and energy. The intra-assay coefficients of variation for all outcomes were less than 3%. The FT mid-IR spectroscope demonstrated high accuracy in measurement of total protein and fat of preterm and term milk with high precision.

  12. Rapid and Sensitive Detection of an Intracellular Pathogen in Human Peripheral Leukocytes with Hybridizing Magnetic Relaxation Nanosensors

    Science.gov (United States)

    Kaittanis, Charalambos; Boukhriss, Hamza; Santra, Santimukul; Naser, Saleh A.; Perez, J. Manuel

    2012-01-01

    Bacterial infections are still a major global healthcare problem. The quick and sensitive detection of pathogens responsible for these infections would facilitate correct diagnosis of the disease and expedite treatment. Of major importance are intracellular slow-growing pathogens that reside within peripheral leukocytes, evading recognition by the immune system and detection by traditional culture methods. Herein, we report the use of hybridizing magnetic nanosensors (hMRS) for the detection of an intracellular pathogen, Mycobacterium avium spp. paratuberculosis (MAP). The hMRS are designed to bind to a unique genomic sequence found in the MAP genome, causing significant changes in the sample’s magnetic resonance signal. Clinically relevant samples, including tissue and blood, were screened with hMRS and results were compared with traditional PCR analysis. Within less than an hour, the hMRS identified MAP-positive samples in a library of laboratory cultures, clinical isolates, blood and homogenized tissues. Comparison of the hMRS with culture methods in terms of prediction of disease state revealed that the hMRS outperformed established culture methods, while being significantly faster (1 hour vs 12 weeks). Additionally, using a single instrument and one nanoparticle preparation we were able to detect the intracellular bacterial target in clinical samples at the genomic and epitope levels. Overall, since the nanoparticles are robust in diverse environmental settings and substantially more affordable than PCR enzymes, the potential clinical and field-based use of hMRS in the multiplexed identification of microbial pathogens and other disease-related biomarkers via a single, deployable instrument in clinical and complex environmental samples is foreseen. PMID:22496916

  13. Rapid and sensitive detection of an intracellular pathogen in human peripheral leukocytes with hybridizing magnetic relaxation nanosensors.

    Directory of Open Access Journals (Sweden)

    Charalambos Kaittanis

    Full Text Available Bacterial infections are still a major global healthcare problem. The quick and sensitive detection of pathogens responsible for these infections would facilitate correct diagnosis of the disease and expedite treatment. Of major importance are intracellular slow-growing pathogens that reside within peripheral leukocytes, evading recognition by the immune system and detection by traditional culture methods. Herein, we report the use of hybridizing magnetic nanosensors (hMRS for the detection of an intracellular pathogen, Mycobacterium avium spp. paratuberculosis (MAP. The hMRS are designed to bind to a unique genomic sequence found in the MAP genome, causing significant changes in the sample's magnetic resonance signal. Clinically relevant samples, including tissue and blood, were screened with hMRS and results were compared with traditional PCR analysis. Within less than an hour, the hMRS identified MAP-positive samples in a library of laboratory cultures, clinical isolates, blood and homogenized tissues. Comparison of the hMRS with culture methods in terms of prediction of disease state revealed that the hMRS outperformed established culture methods, while being significantly faster (1 hour vs 12 weeks. Additionally, using a single instrument and one nanoparticle preparation we were able to detect the intracellular bacterial target in clinical samples at the genomic and epitope levels. Overall, since the nanoparticles are robust in diverse environmental settings and substantially more affordable than PCR enzymes, the potential clinical and field-based use of hMRS in the multiplexed identification of microbial pathogens and other disease-related biomarkers via a single, deployable instrument in clinical and complex environmental samples is foreseen.

  14. Rapidly Escalating Hepcidin and Associated Serum Iron Starvation Are Features of the Acute Response to Typhoid Infection in Humans.

    Directory of Open Access Journals (Sweden)

    Thomas C Darton

    2015-09-01

    Full Text Available Iron is a key pathogenic determinant of many infectious diseases. Hepcidin, the hormone responsible for governing systemic iron homeostasis, is widely hypothesized to represent a key component of nutritional immunity through regulating the accessibility of iron to invading microorganisms during infection. However, the deployment of hepcidin in human bacterial infections remains poorly characterized. Typhoid fever is a globally significant, human-restricted bacterial infection, but understanding of its pathogenesis, especially during the critical early phases, likewise is poorly understood. Here, we investigate alterations in hepcidin and iron/inflammatory indices following experimental human typhoid challenge.Fifty study participants were challenged with Salmonella enterica serovar Typhi and monitored for evidence of typhoid fever. Serum hepcidin, ferritin, serum iron parameters, C-reactive protein (CRP, and plasma IL-6 and TNF-alpha concentrations were measured during the 14 days following challenge. We found that hepcidin concentrations were markedly higher during acute typhoid infection than at baseline. Hepcidin elevations mirrored the kinetics of fever, and were accompanied by profound hypoferremia, increased CRP and ferritin, despite only modest elevations in IL-6 and TNF-alpha in some individuals. During inflammation, the extent of hepcidin upregulation associated with the degree of hypoferremia.We demonstrate that strong hepcidin upregulation and hypoferremia, coincident with fever and systemic inflammation, are hallmarks of the early innate response to acute typhoid infection. We hypothesize that hepcidin-mediated iron redistribution into macrophages may contribute to S. Typhi pathogenesis by increasing iron availability for macrophage-tropic bacteria, and that targeting macrophage iron retention may represent a strategy for limiting infections with macrophage-tropic pathogens such as S. Typhi.

  15. Rapidly Escalating Hepcidin and Associated Serum Iron Starvation Are Features of the Acute Response to Typhoid Infection in Humans.

    Science.gov (United States)

    Darton, Thomas C; Blohmke, Christoph J; Giannoulatou, Eleni; Waddington, Claire S; Jones, Claire; Sturges, Pamela; Webster, Craig; Drakesmith, Hal; Pollard, Andrew J; Armitage, Andrew E

    2015-09-01

    Iron is a key pathogenic determinant of many infectious diseases. Hepcidin, the hormone responsible for governing systemic iron homeostasis, is widely hypothesized to represent a key component of nutritional immunity through regulating the accessibility of iron to invading microorganisms during infection. However, the deployment of hepcidin in human bacterial infections remains poorly characterized. Typhoid fever is a globally significant, human-restricted bacterial infection, but understanding of its pathogenesis, especially during the critical early phases, likewise is poorly understood. Here, we investigate alterations in hepcidin and iron/inflammatory indices following experimental human typhoid challenge. Fifty study participants were challenged with Salmonella enterica serovar Typhi and monitored for evidence of typhoid fever. Serum hepcidin, ferritin, serum iron parameters, C-reactive protein (CRP), and plasma IL-6 and TNF-alpha concentrations were measured during the 14 days following challenge. We found that hepcidin concentrations were markedly higher during acute typhoid infection than at baseline. Hepcidin elevations mirrored the kinetics of fever, and were accompanied by profound hypoferremia, increased CRP and ferritin, despite only modest elevations in IL-6 and TNF-alpha in some individuals. During inflammation, the extent of hepcidin upregulation associated with the degree of hypoferremia. We demonstrate that strong hepcidin upregulation and hypoferremia, coincident with fever and systemic inflammation, are hallmarks of the early innate response to acute typhoid infection. We hypothesize that hepcidin-mediated iron redistribution into macrophages may contribute to S. Typhi pathogenesis by increasing iron availability for macrophage-tropic bacteria, and that targeting macrophage iron retention may represent a strategy for limiting infections with macrophage-tropic pathogens such as S. Typhi.

  16. The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage

    DEFF Research Database (Denmark)

    Christiansen, Stig Hill; Zhang, Xianwei; Juul-Madsen, Kristian

    2017-01-01

    in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell...... membrane deformation. Nanoparticle tracking analysis confirmed the formation of such debris, even at low GA concentrations. Electric cell-substrate impedance sensing measurements also recorded stable alterations in T lymphocytes following such treatment. LL-37 forms oligomers through weak hydrophobic...

  17. Rapid and High-Throughput Detection and Quantitation of Radiation Biomarkers in Human and Nonhuman Primates by Differential Mobility Spectrometry-Mass Spectrometry.

    Science.gov (United States)

    Chen, Zhidan; Coy, Stephen L; Pannkuk, Evan L; Laiakis, Evagelia C; Hall, Adam B; Fornace, Albert J; Vouros, Paul

    2016-10-01

    Radiation exposure is an important public health issue due to a range of accidental and intentional threats. Prompt and effective large-scale screening and appropriate use of medical countermeasures (MCM) to mitigate radiation injury requires rapid methods for determining the radiation dose. In a number of studies, metabolomics has identified small-molecule biomarkers responding to the radiation dose. Differential mobility spectrometry-mass spectrometry (DMS-MS) has been used for similar compounds for high-throughput small-molecule detection and quantitation. In this study, we show that DMS-MS can detect and quantify two radiation biomarkers, trimethyl-L-lysine (TML) and hypoxanthine. Hypoxanthine is a human and nonhuman primate (NHP) radiation biomarker and metabolic intermediate, whereas TML is a radiation biomarker in humans but not in NHP, which is involved in carnitine synthesis. They have been analyzed by DMS-MS from urine samples after a simple strong cation exchange-solid phase extraction (SCX-SPE). The dramatic suppression of background and chemical noise provided by DMS-MS results in an approximately 10-fold reduction in time, including sample pretreatment time, compared with liquid chromatography-mass spectrometry (LC-MS). DMS-MS quantitation accuracy has been verified by validation testing for each biomarker. Human samples are not yet available, but for hypoxanthine, selected NHP urine samples (pre- and 7-d-post 10 Gy exposure) were analyzed, resulting in a mean change in concentration essentially identical to that obtained by LC-MS (fold-change 2.76 versus 2.59). These results confirm the potential of DMS-MS for field or clinical first-level rapid screening for radiation exposure. Graphical Abstract ᅟ.

  18. Rapid multicomponent T2 analysis of the articular cartilage of the human knee joint at 3.0T.

    Science.gov (United States)

    Liu, Fang; Chaudhary, Rajeev; Hurley, Samuel A; Munoz Del Rio, Alejandro; Alexander, Andrew L; Samsonov, Alexey; Block, Walter F; Kijowski, Richard

    2014-05-01

    To determine the feasibility of using multicomponent-driven equilibrium single-shot observation of T1 and T2 (mcDESPOT) for evaluating the human knee joint at 3.0T and to investigate depth-dependent and regional-dependent variations in multicomponent T2 parameters within articular cartilage. mcDESPOT was performed on the knee joint of 10 asymptomatic volunteers at 3.0T. Single-component T2 relaxation time (T2single ), multicomponent T2 relaxation time for water tightly bound to proteoglycan (T2PG ) and bulk water loosely bound to the macromolecular matrix (T2BW ), and fraction of water tightly bound to proteoglycan (FPG ) were measured in eight cartilage subsections and within the superficial and deep layers of patellar cartilage. Statistical analysis was used to investigate depth-dependent and regional-dependent variations in parameters. There was lower (P = 0.001) T2single and T2PG and higher (P Multicomponent T2 parameters of the articular cartilage of the human knee joint can be measured at 3.0T using mcDESPOT and show depth-dependent and regional-dependent variations. Copyright © 2013 Wiley Periodicals, Inc.

  19. Cryopreservation of human oocytes, zygotes, embryos and blastocysts: A comparison study between slow freezing and ultra rapid (vitrification methods

    Directory of Open Access Journals (Sweden)

    Tahani Al-Azawi

    2013-12-01

    Full Text Available Preservation of female genetics is currently done primarily by means of oocyte and embryo cryopreservation. The field has seen much progress during its four-decade history, progress driven predominantly by research in humans. It can also be done by preservation of ovarian tissue or entire ovary for transplantation, followed by oocyte harvesting or natural fertilization. Two basic cryopreservation techniques rule the field, slow-rate freezing, the first to be developed and vitrification which in recent years, has gained a foothold. The slow-rate freezing method previously reported had low survival and pregnancy rates, along with the high cost of cryopreservation. Although there are some recent data indicating better survival rates, cryopreservation by the slow freezing method has started to discontinue. Vitrification of human embryos, especially at early stages, became a more popular alternative to the slow rate freezing method due to reported comparable clinical and laboratory outcomes. In addition, vitrification is relatively simple, requires no expensive programmable freezing equipment, and uses a small amount of liquid nitrogen for freezing. Moreover, oocyte cryopreservation using vitrification has been proposed as a solution to maintain women’s fertility by serving and freezing their oocytes at the optimal time. The aim of this research is to compare slow freezing and vitrification in cryopreservation of oocytes, zygotes, embryos and blastocysts during the last twelve years. Therefore, due to a lot of controversies in this regard, we tried to achieve an exact idea about the subject and the best technique used.

  20. Rapid detection of methylation change at H19 in human imprinting disorders using methylation-sensitive high-resolution melting.

    Science.gov (United States)

    Wojdacz, Tomasz K; Dobrovic, Alexander; Algar, Elizabeth M

    2008-10-01

    Beckwith Wiedemann syndrome (BWS) and Russell Silver syndrome (RS) are growth disorders with opposing epimutations affecting the H19/IGF2 imprinting center at 11p15.5. Overgrowth and tumor risk in BWS is caused by aberrant expression of the paternally expressed, imprinted IGF2 gene, occurring as a consequence of mosaic hypermethylation within the imprinting center, or to mosaic paternal uniparental disomy (UPD). RS is characterized by severe intrauterine growth retardation (IUGR). A subset of RS cases were recently shown to have mosaic hypomethylation within the H19/IGF2 imprinting center, predicted to silence paternally expressed IGF2 in early development. Molecular diagnosis for BWS and RS involves methylation analysis of the H19 locus, enabling discrimination of allelic methylation patterns. In this study, methylation-sensitive high-resolution melting analysis (MS-HRM) was used to analyze methylation within the intergenic region of the H19 locus. A total of 36 samples comprising normal control (11), BWS (19), and RS (six) DNA were analyzed in a blinded study and scored as hypermethylated, normal, or hypomethylated. Results were compared with those derived by methylation-sensitive Southern blotting using the restriction enzymes Rsa I and Hpa II. A total of 100% concordance was obtained for the Southern blotting and MS-HRM scores. A total of three samples with paternal duplication affecting the H19/IGF2 region were scored as equivocal by both methods; however, 33 out of 36 (92%) the samples were unambiguously scored as being hypermethylated, hypomethylated, or normally methylated using MS-HRM. We conclude that MS-HRM is a rapid, cost-effective, and sensitive method for screening mosaic methylation changes at the H19 locus in BWS and RS.

  1. Novel photosensitizers trigger rapid death of malignant human cells and rodent tumor transplants via lipid photodamage and membrane permeabilization.

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    Mikhail M Moisenovich

    Full Text Available BACKGROUND: Apoptotic cascades may frequently be impaired in tumor cells; therefore, the approaches to circumvent these obstacles emerge as important therapeutic modalities. METHODOLOGY/PRINCIPAL FINDINGS: Our novel derivatives of chlorin e(6, that is, its amide (compound 2 and boronated amide (compound 5 evoked no dark toxicity and demonstrated a significantly higher photosensitizing efficacy than chlorin e(6 against transplanted aggressive tumors such as B16 melanoma and M-1 sarcoma. Compound 5 showed superior therapeutic potency. Illumination with red light of mammalian tumor cells loaded with 0.1 µM of 5 caused rapid (within the initial minutes necrosis as determined by propidium iodide staining. The laser confocal microscopy-assisted analysis of cell death revealed the following order of events: prior to illumination, 5 accumulated in Golgi cysternae, endoplasmic reticulum and in some (but not all lysosomes. In response to light, the reactive oxygen species burst was concomitant with the drop of mitochondrial transmembrane electric potential, the dramatic changes of mitochondrial shape and the loss of integrity of mitochondria and lysosomes. Within 3-4 min post illumination, the plasma membrane became permeable for propidium iodide. Compounds 2 and 5 were one order of magnitude more potent than chlorin e(6 in photodamage of artificial liposomes monitored in a dye release assay. The latter effect depended on the content of non-saturated lipids; in liposomes consisting of saturated lipids no photodamage was detectable. The increased therapeutic efficacy of 5 compared with 2 was attributed to a striking difference in the ability of these photosensitizers to permeate through hydrophobic membrane interior as evidenced by measurements of voltage jump-induced relaxation of transmembrane current on planar lipid bilayers. CONCLUSIONS/SIGNIFICANCE: The multimembrane photodestruction and cell necrosis induced by photoactivation of 2 and 5 are

  2. Rapid isolation of gluten-digesting bacteria from human stool and saliva by using gliadin-containing plates

    Science.gov (United States)

    Sarantopoulos, Christos; Ongchangco, Deryn; Sry, Jeremy; Cesario, Thomas

    2014-01-01

    The number of individuals with gluten intolerance has increased dramatically over the last years. To date, the only therapy for gluten intolerance is the complete avoidance of dietary gluten. To sustain a strictly gluten-free diet, however, is very challenging. Therefore, there is need for a non-dietary therapy. Any such treatment must appreciate that the immunogenic part of gluten are gliadin peptides which are poorly degraded by the enzymes of the gastrointestinal tract. Probiotic therapy and oral enzyme therapy containing gluten-degrading bacteria (GDB) and their gliadin-digesting enzymes are possible new approaches for the treatment of gluten intolerance, however effectively isolating GDB for these treatments is problematic. The goal of this study was to develop an easy technique to isolate GDB rapidly and efficiently with the hope it might lead to newer ways of developing either probiotics or traditional medicines to treat gluten intolerance. Several researchers have already isolated successfully GDB by using gluten minimal or limited agar plates. Although these plates can be used to isolate bacteria which can tolerate gluten, further assays are needed to investigate if the same bacteria can also digest gluten. The agar plates we developed can detect bacteria which cannot only tolerate gluten but are able to digest it as well. Therefore, we were able to combine two steps into one step. Using such technologies, we were able to isolate five GDB from saliva and stool, and identified three bacterial reference strains with gluten-degrading activity. The technique we developed to isolate bacteria with gluten-degrading activity is fast, effective, and easy to use. The GDB isolated by our technology could have potential as part of a probiotic or enzymatic therapy for people with gluten intolerance. PMID:25519429

  3. Rapid isolation of gluten-digesting bacteria from human stool and saliva by using gliadin-containing plates.

    Science.gov (United States)

    Berger, Martina; Sarantopoulos, Christos; Ongchangco, Deryn; Sry, Jeremy; Cesario, Thomas

    2015-07-01

    The number of individuals with gluten intolerance has increased dramatically over the last years. To date, the only therapy for gluten intolerance is the complete avoidance of dietary gluten. To sustain a strictly gluten-free diet, however, is very challenging. Therefore, there is need for a non-dietary therapy. Any such treatment must appreciate that the immunogenic part of gluten are gliadin peptides which are poorly degraded by the enzymes of the gastrointestinal tract. Probiotic therapy and oral enzyme therapy containing gluten-degrading bacteria (GDB) and their gliadin-digesting enzymes are possible new approaches for the treatment of gluten intolerance, however effectively isolating GDB for these treatments is problematic. The goal of this study was to develop an easy technique to isolate GDB rapidly and efficiently with the hope it might lead to newer ways of developing either probiotics or traditional medicines to treat gluten intolerance. Several researchers have already isolated successfully GDB by using gluten minimal or limited agar plates. Although these plates can be used to isolate bacteria which can tolerate gluten, further assays are needed to investigate if the same bacteria can also digest gluten. The agar plates we developed can detect bacteria which cannot only tolerate gluten but are able to digest it as well. Therefore, we were able to combine two steps into one step. Using such technologies, we were able to isolate five GDB from saliva and stool, and identified three bacterial reference strains with gluten-degrading activity. The technique we developed to isolate bacteria with gluten-degrading activity is fast, effective, and easy to use. The GDB isolated by our technology could have potential as part of a probiotic or enzymatic therapy for people with gluten intolerance. © 2014 by the Society for Experimental Biology and Medicine.

  4. Comparison of the Diagnostic Accuracy of Three Rapid Tests for the Serodiagnosis of Hepatic Cystic Echinococcosis in Humans.

    Science.gov (United States)

    Tamarozzi, Francesca; Covini, Ilaria; Mariconti, Mara; Narra, Roberta; Tinelli, Carmine; De Silvestri, Annalisa; Manzoni, Federica; Casulli, Adriano; Ito, Akira; Neumayr, Andreas; Brunetti, Enrico

    2016-02-01

    The diagnosis of cystic echinococcosis (CE) is based primarily on imaging, in particular with ultrasound for abdominal CE, complemented by serology when imaging results are unclear. In rural endemic areas, where expertise in ultrasound may be scant and conventional serology techniques are unavailable due to lack of laboratory equipment, Rapid Diagnostic Tests (RDTs) are appealing. We evaluated the diagnostic accuracy of 3 commercial RDTs for the diagnosis of hepatic CE. Sera from 59 patients with single hepatic CE cysts in well-defined ultrasound stages (gold standard) and 25 patients with non-parasitic cysts were analyzed by RDTs VIRapid HYDATIDOSIS (Vircell, Spain), Echinococcus DIGFA (Unibiotest, China), ADAMU-CE (ICST, Japan), and by RIDASCREEN Echinococcus IgG ELISA (R-Biopharm, Germany). Sensitivity, specificity and ROC curves were compared with McNemar and t-test. For VIRapid and DIGFA, correlation between semiquantitative results and ELISA OD values were evaluated by Spearman's coefficient. Reproducibility was assessed on 16 randomly selected sera with Cohen's Kappa coefficient. Sensitivity and Specificity of VIRapid (74%, 96%) and ADAMU-CE (57%, 100%) did not differ from ELISA (69%, 96%) while DIGFA (72%, 72%) did (p = 0.045). ADAMU-CE was significantly less sensitive in the diagnosis of active cysts (p = 0.019) while DIGFA was significantly less specific (p = 0.014) compared to ELISA. All tests were poorly sensitive in diagnosing inactive cysts (33.3% ELISA and ADAMU-CE, 42.8% DIGFA, 47.6% VIRapid). The reproducibility of all RDTs was good-very good. Band intensity of VIRapid and DIGFA correlated with ELISA OD values (r = 0.76 and r = 0.79 respectively, p<0.001). RDTs may be useful in resource-poor settings to complement ultrasound diagnosis of CE in uncertain cases. VIRapid test appears to perform best among the examined kits, but all tests are poorly sensitive in the presence of inactive cysts, which may pose problems with accurate diagnosis.

  5. A computer vision system for rapid search inspired by surface-based attention mechanisms from human perception.

    Science.gov (United States)

    Mohr, Johannes; Park, Jong-Han; Obermayer, Klaus

    2014-12-01

    Humans are highly efficient at visual search tasks by focusing selective attention on a small but relevant region of a visual scene. Recent results from biological vision suggest that surfaces of distinct physical objects form the basic units of this attentional process. The aim of this paper is to demonstrate how such surface-based attention mechanisms can speed up a computer vision system for visual search. The system uses fast perceptual grouping of depth cues to represent the visual world at the level of surfaces. This representation is stored in short-term memory and updated over time. A top-down guided attention mechanism sequentially selects one of the surfaces for detailed inspection by a recognition module. We show that the proposed attention framework requires little computational overhead (about 11 ms), but enables the system to operate in real-time and leads to a substantial increase in search efficiency. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Human resource development and capacity-building during China’s rapid scale-up of methadone maintenance treatment services

    Science.gov (United States)

    Li, Jianhua; Wang, Changhe; McGoogan, Jennifer M; Rou, Keming; Bulterys, Marc

    2013-01-01

    Abstract Problem China’s National Methadone Maintenance Treatment Programme (MMT) has expanded from eight clinics serving approximately 1000 clients to 738 clinics that have served more than 340 000 clients cumulatively in only 8 years. This has created an enormous demand for trained providers. Approach Human resource development and capacity building efforts have been conducted in China’s National MMT Programme to create a supply of providers trained in administering MMT for opioid dependence. Local setting From 2004 to 2007, China’s National MMT Programme faced several problems: inappropriately low methadone doses, poor compliance, high concurrent drug use and high drop-out rates among clients, and little experience, little training and high turnover rates among providers. Relevant changes Training programmes for individual providers and their trainers were redeveloped and expanded in 2008. Although programme performance metrics show an increase in patients’ annual mean duration in treatment (93 days in 2004 versus 238 days in 2011), the increase in their mean daily methadone dose (from 47.2 mg in 2004 to 58.6 mg in 2011) is modest. Lessons learnt Some of the problems that can arise during the development, launch and scale-up of a major national public health effort, such as China’s National MMT Programme, cannot be foreseen. Key to the programme’s success so far have been the strong commitment on the part of China’s government and the optimism and pragmatism of programme managers. Human resources development and capacity-building during scale-up have contributed to improved service quality in MMT treatment clinics and are critical to long-term success. PMID:23554525

  7. Human resource development and capacity-building during China's rapid scale-up of methadone maintenance treatment services.

    Science.gov (United States)

    Li, Jianhua; Wang, Changhe; McGoogan, Jennifer M; Rou, Keming; Bulterys, Marc; Wu, Zunyou

    2013-02-01

    China's National Methadone Maintenance Treatment Programme (MMT) has expanded from eight clinics serving approximately 1000 clients to 738 clinics that have served more than 340 000 clients cumulatively in only 8 years. This has created an enormous demand for trained providers. Human resource development and capacity building efforts have been conducted in China's National MMT Programme to create a supply of providers trained in administering MMT for opioid dependence. From 2004 to 2007, China's National MMT Programme faced several problems: inappropriately low methadone doses, poor compliance, high concurrent drug use and high drop-out rates among clients, and little experience, little training and high turnover rates among providers. Training programmes for individual providers and their trainers were redeveloped and expanded in 2008. Although programme performance metrics show an increase in patients' annual mean duration in treatment (93 days in 2004 versus 238 days in 2011), the increase in their mean daily methadone dose (from 47.2 mg in 2004 to 58.6 mg in 2011) is modest. Some of the problems that can arise during the development, launch and scale-up of a major national public health effort, such as China's National MMT Programme, cannot be foreseen. Key to the programme's success so far have been the strong commitment on the part of China's government and the optimism and pragmatism of programme managers. Human resources development and capacity-building during scale-up have contributed to improved service quality in MMT treatment clinics and are critical to long-term success.

  8. Rapid determination of myosin heavy chain expression in rat, mouse, and human skeletal muscle using multicolor immunofluorescence analysis.

    Directory of Open Access Journals (Sweden)

    Darin Bloemberg

    Full Text Available Skeletal muscle is a heterogeneous tissue comprised of fibers with different morphological, functional, and metabolic properties. Different muscles contain varying proportions of fiber types; therefore, accurate identification is important. A number of histochemical methods are used to determine muscle fiber type; however, these techniques have several disadvantages. Immunofluorescence analysis is a sensitive method that allows for simultaneous evaluation of multiple MHC isoforms on a large number of fibers on a single cross-section, and offers a more precise means of identifying fiber types. In this investigation we characterized pure and hybrid fiber type distribution in 10 rat and 10 mouse skeletal muscles, as well as human vastus lateralis (VL using multicolor immunofluorescence analysis. In addition, we determined fiber type-specific cross-sectional area (CSA, succinate dehydrogenase (SDH activity, and α-glycerophosphate dehydrogenase (GPD activity. Using this procedure we were able to easily identify pure and hybrid fiber populations in rat, mouse, and human muscle. Hybrid fibers were identified in all species and made up a significant portion of the total population in some rat and mouse muscles. For example, rat mixed gastrocnemius (MG contained 12.2% hybrid fibers whereas mouse white tibialis anterior (WTA contained 12.1% hybrid fibers. Collectively, we outline a simple and time-efficient method for determining MHC expression in skeletal muscle of multiple species. In addition, we provide a useful resource of the pure and hybrid fiber type distribution, fiber CSA, and relative fiber type-specific SDH and GPD activity in a number of rat and mouse muscles.

  9. Rapid Profiling of Bovine and Human Milk Gangliosides by Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.

    Science.gov (United States)

    Lee, Hyeyoung; An, Hyun Joo; Lerno, Larry A; German, J Bruce; Lebrilla, Carlito B

    2011-08-15

    Gangliosides are anionic glycosphingolipids widely distributed in vertebrate tissues and fluids. Their structural and quantitative expression patterns depend on phylogeny and are distinct down to the species level. In milk, gangliosides are exclusively associated with the milk fat globule membrane. They may participate in diverse biological processes but more specifically to host-pathogen interactions. However, due to the molecular complexities, the analysis needs extensive sample preparation, chromatographic separation, and even chemical reaction, which makes the process very complex and time-consuming. Here, we describe a rapid profiling method for bovine and human milk gangliosides employing matrix-assisted desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Prior to the analyses of biological samples, milk ganglioside standards GM3 and GD3 fractions were first analyzed in order to validate this method. High mass accuracy and high resolution obtained from MALDI FTICR MS allow for the confident assignment of chain length and degree of unsaturation of the ceramide. For the structural elucidation, tandem mass spectrometry (MS/MS), specifically as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) were employed. Complex ganglioside mixtures from bovine and human milk were further analyzed with this method. The samples were prepared by two consecutive chloroform/methanol extraction and solid phase extraction. We observed a number of differences between bovine milk and human milk. The common gangliosides in bovine and human milk are NeuAc-NeuAc-Hex-Hex-Cer (GD3) and NeuAc-Hex-Hex-Cer (GM3); whereas, the ion intensities of ganglioside species are different between two milk samples. Kendrick mass defect plot yields grouping of ganglioside peaks according to their structural similarities. Gangliosides were further probed by tandem MS to confirm the compositional and structural assignments

  10. Comparison of the Diagnostic Accuracy of Three Rapid Tests for the Serodiagnosis of Hepatic Cystic Echinococcosis in Humans.

    Directory of Open Access Journals (Sweden)

    Francesca Tamarozzi

    2016-02-01

    Full Text Available The diagnosis of cystic echinococcosis (CE is based primarily on imaging, in particular with ultrasound for abdominal CE, complemented by serology when imaging results are unclear. In rural endemic areas, where expertise in ultrasound may be scant and conventional serology techniques are unavailable due to lack of laboratory equipment, Rapid Diagnostic Tests (RDTs are appealing.We evaluated the diagnostic accuracy of 3 commercial RDTs for the diagnosis of hepatic CE. Sera from 59 patients with single hepatic CE cysts in well-defined ultrasound stages (gold standard and 25 patients with non-parasitic cysts were analyzed by RDTs VIRapid HYDATIDOSIS (Vircell, Spain, Echinococcus DIGFA (Unibiotest, China, ADAMU-CE (ICST, Japan, and by RIDASCREEN Echinococcus IgG ELISA (R-Biopharm, Germany. Sensitivity, specificity and ROC curves were compared with McNemar and t-test. For VIRapid and DIGFA, correlation between semiquantitative results and ELISA OD values were evaluated by Spearman's coefficient. Reproducibility was assessed on 16 randomly selected sera with Cohen's Kappa coefficient. Sensitivity and Specificity of VIRapid (74%, 96% and ADAMU-CE (57%, 100% did not differ from ELISA (69%, 96% while DIGFA (72%, 72% did (p = 0.045. ADAMU-CE was significantly less sensitive in the diagnosis of active cysts (p = 0.019 while DIGFA was significantly less specific (p = 0.014 compared to ELISA. All tests were poorly sensitive in diagnosing inactive cysts (33.3% ELISA and ADAMU-CE, 42.8% DIGFA, 47.6% VIRapid. The reproducibility of all RDTs was good-very good. Band intensity of VIRapid and DIGFA correlated with ELISA OD values (r = 0.76 and r = 0.79 respectively, p<0.001.RDTs may be useful in resource-poor settings to complement ultrasound diagnosis of CE in uncertain cases. VIRapid test appears to perform best among the examined kits, but all tests are poorly sensitive in the presence of inactive cysts, which may pose problems with accurate diagnosis.

  11. Development and validation of sensitive and rapid UPLC-MS/MS method for quantitative determination of daclatasvir in human plasma: Application to a bioequivalence study.

    Science.gov (United States)

    Rezk, Mamdouh R; Bendas, Ehab R; Basalious, Emad B; Karim, Iman A

    2016-09-05

    A rapid and sensitive UPLC-MS/MS method was developed and validated for determination of daclatasvir (DAC) in human plasma using sofosbuvir (SOF) as an internal standard (IS). The Xevo TQD LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Precipitation with acetonitrile was used in sample preparation. The prepared samples were chromatographed on Acquity UPLC HSS C18 (50×2.1mm, 1.8μm) column by pumping 10mM ammonium formate (pH 3.5) and acetonitrile in an isocratic mode at a flow rate of 0.30ml/min. Method validation was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 5-4000ng/ml for DAC. The intra-day and inter-day precision and accuracy results were within the acceptable limits. A very short run time of 1.2min made it possible to analyze more than 500 human plasma samples per day. The wider range of quantification of DAC allowed the applicability of the developed method for its determination in a bioequivalence study in human volunteers. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Rapid MALDI-TOF-MS analysis in the study of interaction between whole bacterial cells and human target molecules: binding of Bifidobacterium to human plasminogen.

    Science.gov (United States)

    Candela, Marco; Fiori, Jessica; Dipalo, Samuele; Naldi, Marina; Gotti, Roberto; Brigidi, Patrizia

    2008-06-01

    MALDI-TOF (Matrix Assisted Laser Desorption Ionization-Time of Flight)-mass spectrometry has been applied, for the first time, in the investigation of whole Bifidobacterium cells-host target proteins interaction. In particular, by means of this technique, a dose dependent human plasminogen-binding activity has been shown for Bifidobacterium. The involvement of lysine binding sites on the bacterial cell surface has been proved. The obtained result was found to be consistent with that from well-established standard methodologies, thus the proposed MALDI-TOF approach has the potential to enter as a fast alternative method in the field of biorecognition studies involving in bacterial cells and proteins of human origin.

  13. Transport of H2S and HS(-) across the human red blood cell membrane: rapid H2S diffusion and AE1-mediated Cl(-)/HS(-) exchange.

    Science.gov (United States)

    Jennings, Michael L

    2013-11-01

    The rates of H2S and HS(-) transport across the human erythrocyte membrane were estimated by measuring rates of dissipation of pH gradients in media containing 250 μM H2S/HS(-). Net acid efflux is caused by H2S/HS(-) acting analogously to CO2/HCO3(-) in the Jacobs-Stewart cycle. The steps are as follows: 1) H2S efflux through the lipid bilayer and/or a gas channel, 2) extracellular H2S deprotonation, 3) HS(-) influx in exchange for Cl(-), catalyzed by the anion exchange protein AE1, and 4) intracellular HS(-) protonation. Net acid transport by the Cl(-)/HS(-)/H2S cycle is more efficient than by the Cl(-)/HCO3(-)/CO2 cycle because of the rapid H2S-HS(-) interconversion in cells and medium. The rates of acid transport were analyzed by solving the mass flow equations for the cycle to produce estimates of the HS(-) and H2S transport rates. The data indicate that HS(-) is a very good substrate for AE1; the Cl(-)/HS(-) exchange rate is about one-third as rapid as Cl(-)/HCO3(-) exchange. The H2S permeability coefficient must also be high (>10(-2) cm/s, half time <0.003 s) to account for the pH equilibration data. The results imply that H2S and HS(-) enter erythrocytes very rapidly in the microcirculation of H2S-producing tissues, thereby acting as a sink for H2S and lowering the local extracellular concentration, and the fact that HS(-) is a substrate for a Cl(-)/HCO3(-) exchanger indicates that some effects of exogenous H2S/HS(-) may not result from a regulatory role of H2S but, rather, from net acid flux by H2S and HS(-) transport in a Jacobs-Stewart cycle.

  14. Simple and rapid human papillomavirus genotyping method by restriction fragment length polymorphism analysis with two restriction enzymes.

    Science.gov (United States)

    Chen, Linghan; Watanabe, Ken; Haruyama, Takahiro; Kobayashi, Nobuyuki

    2013-07-01

    Cervical cancer, the third most common cancer that affects women worldwide, is caused by the human papillomavirus (HPV) and is treatable when detected at an early stage. To date, more than 100 different HPV types have been described, and the development of simple, low-cost, and accurate methods to distinguish HPV genotypes is highly warranted. In this study, an HPV genotyping assay based on polymerase chain reaction (PCR) was evaluated. This method involved the use of MY09/11 primers followed by restriction fragment length polymorphism (RFLP) analysis with the restriction enzymes HpyCH4V and NlaIII. Cervical specimens preserved using CytoRich Blue fluid were collected from 1,134 female volunteers for HPV detection, and 1,111 valid samples were amplified using PCR. The PCR method was sensitive enough to detect 25 copies of HPV18, and three copies of HPV16. Out of 202 PCR-positive samples, HPV genotypes were determined in 189 samples (93.6%) by this RFLP method. Results were then evaluated further by capillary sequencing method. Concordant results between the two tests were as high as 96.0%. Thirteen samples, which tested negative with RFLP, were verified as non-specific amplifications with PCR. In conclusion, this PCR-RFLP method using restriction enzymes HpyCH4V and NlaIII is simple, non-labor intensive, and is applicable for the inexpensive determination of HPV genotypes in clinical samples. Copyright © 2013 Wiley Periodicals, Inc.

  15. The Calcium-Sensing Receptor Is Necessary for the Rapid Development of Hypercalcemia in Human Lung Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Gwendolen Lorch

    2011-05-01

    Full Text Available The calcium-sensing receptor (CaR is responsible for the regulation of extracellular calcium (Ca2+o homeostasis. CaR activation has been shown to increase proliferation in several cancer cell lines; however, its presence or function has never been documented in lung cancer. We report that Ca2+o-activated CaR results in MAPK-mediated stimulation of parathyroid hormone-related protein (PTHrP production in human lung squamous cell carcinoma (SCC lines and humoral hypercalcemia of malignancy (HHM in vivo. Furthermore, a single nucleotide polymorphism in CaR identified from a hypercalcemia-inducing lung SCC reduced the receptor's activation threshold leading to increased PTHrP expression and secretion. Increasing the expression of either wild-type CaR or a CaR variant with a single nucleotide polymorphism in the cytoplasmic domain was both necessary and sufficient for lung SCC to induce HHM. Because lung cancer patients who frequently develop HHM and PTHrP expression in lung cancer has been only partially explained, the significance of our findings indicates that CaR variants may provide a positive feedback between PTHrP and calcium and result in the syndrome of HHM.

  16. Rapid oriented fibril formation of fish scale collagen facilitates early osteoblastic differentiation of human mesenchymal stem cells.

    Science.gov (United States)

    Matsumoto, Rena; Uemura, Toshimasa; Xu, Zhefeng; Yamaguchi, Isamu; Ikoma, Toshiyuki; Tanaka, Junzo

    2015-08-01

    We studied the effect of fibril formation of fish scale collagen on the osteoblastic differentiation of human mesenchymal stem cells (hMSCs). We found that hMSCs adhered easily to tilapia scale collagen, which remarkably accelerated the early stage of osteoblastic differentiation in hMSCs during in vitro cell culture. Osteoblastic markers such as ALP activity, osteopontin, and bone morphogenetic protein 2 were markedly upregulated when the hMSCs were cultured on a tilapia collagen surface, especially in the early osteoblastic differentiation stage. We hypothesized that this phenomenon occurs due to specific fibril formation of tilapia collagen. Thus, we examined the time course of collagen fibril formation using high-speed atomic force microscopy. Moreover, to elucidate the effect of the orientation of fibril formation on the differentiation of hMSCs, we measured ALP activity of hMSCs cultured on two types of tilapia scale collagen membranes with different degrees of fibril formation. The ALP activity in hMSCs cultured on a fibrous collagen membrane was significantly higher than on a non-fibrous collagen membrane even before adding osteoblastic differentiation medium. These results showed that the degree of the fibril formation of tilapia collagen was essential for the osteoblastic differentiation of hMSCs. © 2014 Wiley Periodicals, Inc.

  17. Rapid disaster victim identification in mass fatality incidents: decision-support tool to facilitate human remains identification.

    Science.gov (United States)

    de Cosmo, Sergio; Barbera, Joseph A

    2012-10-01

    A quantitative decision-support tool (DST), using a combination of selected human physical attributes as identification elements, was developed to facilitate body identification in mass fatality incidents, particularly in settings with limited availability of technological resources and forensic expertise. To construct the DST, the external biological attributes of interest were first selected. A process was then developed to guide collection of the selected categories of attributes and record them into objective antemortem (AM) and postmortem (PM) records. Finally, a framework for assessing the similarity between confronting PM-AM attribute records was established. The DST evaluates the similarities between each set of like attributes in the AM and PM records being compared. It then computes an overall similarity score for each evaluated AM record that was compared to a selected PM record. The AM record with the highest score represents the highest probable match, with the PM file selected for the comparison. Multiple simulations across a range of mass fatality situations demonstrated the effectiveness of the DST in the experimental setting. The developed DST may provide authorities with a method for expediting body identification without completely eliminating any missing person file from consideration. Under specific circumstances, this method may reduce the need for technologically sophisticated forensic identification techniques (eg, dental records, fingerprints, and DNA). At a minimum, it should facilitate the efficiency of the current technological matching process.

  18. Comparative evaluation of TRI-DOT Rapid HIV test with fourth-generation ELISA for the detection of human immunodeficiency virus.

    Science.gov (United States)

    Sudha, T; Teja, V D; Gopal, M; Rajesh, M; Lakshmi, V

    2005-10-01

    This study evaluated the TRI-DOT Rapid HIV test for the early detection of human immunodeficiency virus (HIV) infection in comparison with a fourth-generation ELISA (Vironostika HIV Uniform II). Of 23,609 sera, seven (0.03%) gave discordant results. Six of these were reactive only by the fourth-generation assay and were p24 antigen-positive by VIDAS DUO, Western blot and qualitative RT-PCR tests. The remaining discordant serum was considered to be false-positive by the TRI-DOT assay, as it was negative by repeat ELISA and Western blot tests. The sensitivity and specificity of the TRI-DOT test were 99.48% and 99.99%, respectively, compared with the fourth-generation ELISA.

  19. Rapid measurement of 8-oxo-7,8-dihydro-2'-deoxyguanosine in human biological matrices using ultra-high-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Lam, Patricia M W; Mistry, Vilas; Marczylo, Timothy H; Konje, Justin C; Evans, Mark D; Cooke, Marcus S

    2012-05-15

    Interaction of reactive oxygen species with DNA results in a variety of modifications, including 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), which has been extensively studied as a biomarker of oxidative stress. Oxidative stress is implicated in a number of pathophysiological processes relevant to obstetrics and gynecology; however, there is a lack of understanding as to the precise role of oxidative stress in these processes. We aimed to develop a rapid, validated assay for the accurate quantification of 8-oxodG in human urine using solid-phase extraction and ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and then investigate the levels of 8-oxodG in several fluids of interest to obstetrics and gynecology. Using UHPLC-MS/MS, 8-oxodG eluted after 3.94 min with an RSD for 15 injections of 0.07%. The method was linear between 0.95 and 95 nmol/L with LOD and LOQ of 5 and 25 fmol on-column, respectively. Accuracy and precision were 98.7-101.0 and seminal plasma>amniotic fluid>plasma>serum>peritoneal fluid, and it was not detected in saliva. Urine concentrations normalized to creatinine (n=15) ranged between 0.55 and 1.95 pmol/μmol creatinine. We describe, for the first time, 8-oxodG concentrations in human seminal plasma, peritoneal fluid, amniotic fluid, and breast milk, as well as in urine, plasma, and serum, using a rapid UHPLC-MS/MS method that will further facilitate biomonitoring of oxidative stress. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Simple and rapid LC-MS/MS method for the absolute determination of cetuximab in human serum using an immobilized trypsin.

    Science.gov (United States)

    Shibata, Kaito; Naito, Takafumi; Okamura, Jun; Hosokawa, Seiji; Mineta, Hiroyuki; Kawakami, Junichi

    2017-11-30

    Proteomic approaches using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) without an immunopurification technique have not been applied to the determination of serum cetuximab. This study developed a simple and rapid LC-MS/MS method for the absolute determination of cetuximab in human serum and applied it to clinical settings. Surrogate peptides derived from cetuximab digests were selected using a Fourier transform mass spectrometer. Reduced-alkylated serum cetuximab without immunopurification was digested for 20minutes using immobilized trypsin, and the digestion products were purified by solid-phase extraction. The LC-MS/MS was run in positive ion multiple reaction monitoring mode. This method was applied to the determination of serum samples in head and neck cancer patients treated with cetuximab. The chromatographic run time was 10minutes and no peaks interfering with surrogate peptides in serum digestion products were observed. The calibration curve of absolute cetuximab in serum was linear over the concentration range of 4-200μg/mL. The lower limit of quantification of cetuximab in human serum was 4μg/mL. The intra-assay and inter-assay precision and accuracy were less than 13.2% and 88.0-100.7%, respectively. The serum concentration range of cetuximab was 19-140μg/mL in patients. The serum cetuximab concentrations in LC-MS/MS were correlated with those in ELISA (r=0.899, P <0.01) and the mean bias was 1.5% in cancer patients. In conclusion, the present simple and rapid method with acceptable analytical performance can be helpful for evaluating the absolute concentration of serum cetuximab in clinical settings. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Mitochondrial adaptations to high-volume exercise training are rapidly reversed after a reduction in training volume in human skeletal muscle.

    Science.gov (United States)

    Granata, Cesare; Oliveira, Rodrigo S F; Little, Jonathan P; Renner, Kathrin; Bishop, David J

    2016-10-01

    Increased mitochondrial content and respiration have both been reported after exercise training. However, no study has directly compared how different training volumes influence mitochondrial respiration and markers of mitochondrial biogenesis. Ten healthy men performed high-intensity interval cycling during 3 consecutive training phases; 4 wk of normal-volume training (NVT; 3/wk), followed by 20 d of high-volume training (HVT; 2/d) and 2 wk of reduced-volume training (RVT; 5 sessions). Resting biopsy samples (vastus lateralis) were obtained at baseline and after each phase. No mitochondrial parameter changed after NVT. After HVT, mitochondrial respiration and citrate synthase activity (∼40-50%), as well as the protein content of electron transport system (ETS) subunits (∼10-40%), and that of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), NRF1, mitochondrial transcription factor A (TFAM), PHF20, and p53 (∼65-170%) all increased compared to baseline; mitochondrial specific respiration remained unchanged. After RVT, all the mitochondrial parameters measured except citrate synthase activity (∼36% above initial) were not significantly different compared to baseline (all P > 0.05). Our findings demonstrate that training volume is an important determinant of training-induced mitochondrial adaptations and highlight the rapid reversibility of human skeletal muscle to a reduction in training volume.-Granata, C., Oliveira, R. S. F., Little, J. P., Renner, K., Bishop, D. J. Mitochondrial adaptations to high-volume exercise training are rapidly reversed after a reduction in training volume in human skeletal muscle. © FASEB.

  2. Profuse color-evolution-based fluorescent test paper sensor for rapid and visual monitoring of endogenous Cu(2+) in human urine.

    Science.gov (United States)

    Cai, Yueqing; You, Junhui; You, Zhengyi; Dong, Fang; Du, Shuhu; Zhang, Liying

    2018-01-15

    The fluorescent paper for colorimetric detection of metal ions has been widely fabricated using various sensing probes, but it still remains an elusive task to design a test paper with multicolor variation with target dosages for accurate determination. Herein, we report a profuse color-evolution-based fluorescent test paper sensor for rapid and visual monitoring of Cu(2+) in human urine by printing tricolor probe onto filter paper. The tricolor probe consists of blue-emission carbon dots (bCDs), green-emission quantum dots (gQDs) and red-emission quantum dots (rQDs), which is based on the principle that the fluorescence of gQDs and rQDs are simultaneously quenched by Cu(2+), whereas the bCDs as the photostable internal standard is insensitive to Cu(2+). Upon the addition of different amounts of Cu(2+), the ratiometric fluorescence intensity of the tricolor probe continuously varied, leading to color changes from shallow pink to blue with a detection limit of 1.3nM. When the tricolor probe solution was printed onto a sheet of filter paper, as-obtained test paper displayed a more profuse color evolution from shallow pink to light salmon to dark orange to olive drab to dark olive green to slate blue to royal blue and to final dark blue with the increase of Cu(2+) concentration compared with dual-color probe-based test paper, and dosage scale as low as 6.0nM was clearly discriminated. The sensing test paper is simple, rapid and inexpensive, and serves as a visual platform for ultrasensitive monitoring of endogenous Cu(2+) in human urine. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. A rapid and validated HPLC method to quantify racecadotril metabolite, thiorphan, in human plasma using solid-phase extraction.

    Science.gov (United States)

    Xu, Fan; Yang, Lingli; Xu, Guili

    2008-01-01

    A HPLC method with UV detection was developed and validated for the determination of thiorphan in human plasma. Nevirapine was used as the internal standard. Separation was performed by a Waters sunfire C18 reversed-phase column maintained at 35 degrees C. The mobile phase was a mixture of 0.05 M phosphate buffer with the pH adjusted to 2.6 and acetonitrile (74:26, v/v) at a flow rate of 1.0 mL/min. The UV detector was set at 210 nm. An original pre-treatment of plasma samples was developed, based on solid-phase extraction (SPE) with solid-phase extraction cartridges (Oasis HLB 3 mL, 60 mg). The extraction recovery for plasma samples of thiorphan at 0.1, 0.4 and 2.0 microg/mL was 93.5%, 98.2% and 97.8%, respectively. The calibration curve was linear with the correlation coefficient (r) above 0.9998. Linearity was verified over the range of 0.05-4 microg/mL thiorphan in plasma. The limit of quantification (LOQ) is 0.05 microg/mL. The mean accuracy was 92.7-99.6%. The coefficient of variation (precision) in the within- and between-batch was 2.2-8.4% and 4.1-8.1%, respectively. This method is simple, economical and specific, and has been used successfully in a pharmacokinetic study of thiorphan.

  4. Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP in blood samples

    Directory of Open Access Journals (Sweden)

    Anthony Claudia N

    2011-07-01

    Full Text Available Abstract Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plasmodium ssrRNA genes has been described as the most sensitive and specific method for Plasmodium detection. However, this method is costly and requires trained personnel for its implementation. Loop-mediated isothermal amplification (LAMP, a novel nucleic acid amplification method was developed for the clinical detection of P. knowlesi. The sensitivity and specificity of LAMP was evaluated in comparison to the results obtained via microscopic examination and nested PCR. Methods LAMP assay was developed based on P. knowlesi genetic material targeting the apical membrane antigen-1 (AMA-1 gene. The method uses six primers that recognize eight regions of the target DNA and it amplifies DNA within an hour under isothermal conditions (65°C in a water-bath. Results LAMP is highly sensitive with the detection limit as low as ten copies for AMA-1. LAMP detected malaria parasites in all confirm cases (n = 13 of P. knowlesi infection (sensitivity, 100% and none of the negative samples (specificity, 100% within an hour. LAMP demonstrated higher sensitivity compared to nested PCR by successfully detecting a sample with very low parasitaemia ( Conclusion With continuous efforts in the optimization of this assay, LAMP may provide a simple and reliable test for detecting P. knowlesi malaria parasites in areas where malaria is prevalent.

  5. Gene expression in blood changes rapidly in neutrophils and monocytes after ischemic stroke in humans: a microarray study.

    Science.gov (United States)

    Tang, Yang; Xu, Huichun; Du, XinLi; Lit, Lisa; Walker, Wynn; Lu, Aigang; Ran, Ruiqiong; Gregg, Jeffrey P; Reilly, Melinda; Pancioli, Art; Khoury, Jane C; Sauerbeck, Laura R; Carrozzella, Janice A; Spilker, Judith; Clark, Joseph; Wagner, Kenneth R; Jauch, Edward C; Chang, Dongwoo J; Verro, Piero; Broderick, Joseph P; Sharp, Frank R

    2006-08-01

    Ischemic brain and peripheral white blood cells release cytokines, chemokines and other molecules that activate the peripheral white blood cells after stroke. To assess gene expression in these peripheral white blood cells, whole blood was examined using oligonucleotide microarrays in 15 patients at 2.4+/-0.5, 5 and 24 h after onset of ischemic stroke and compared with control blood samples. The 2.4-h blood samples were drawn before patients were treated either with tissue-type plasminogen activator (tPA) alone or with tPA plus Eptifibatide (the Combination approach to Lysis utilizing Eptifibatide And Recombinant tPA trial). Most genes induced in whole blood at 2 to 3 h were also induced at 5 and 24 h. Separate studies showed that the genes induced at 2 to 24 h after stroke were expressed mainly by polymorphonuclear leukocytes and to a lesser degree by monocytes. These genes included: matrix metalloproteinase 9; S100 calcium-binding proteins P, A12 and A9; coagulation factor V; arginase I; carbonic anhydrase IV; lymphocyte antigen 96 (cluster of differentiation (CD)96); monocarboxylic acid transporter (6); ets-2 (erythroblastosis virus E26 oncogene homolog 2); homeobox gene Hox 1.11; cytoskeleton-associated protein 4; N-formylpeptide receptor; ribonuclease-2; N-acetylneuraminate pyruvate lyase; BCL6; glycogen phosphorylase. The fold change of these genes varied from 1.6 to 6.8 and these 18 genes correctly classified 10/15 patients at 2.4 h, 13/15 patients at 5 h and 15/15 patients at 24 h after stroke. These data provide insights into the inflammatory responses after stroke in humans, and should be helpful in diagnosis, understanding etiology and pathogenesis, and guiding acute treatment and development of new treatments for stroke.

  6. Study of Cholesterol Repletion Effect on Nanomechanical Properties of Human Umbilical Vein Endothelial Cell Via Rapid Broadband Atomic Force Microscopy.

    Science.gov (United States)

    Yan, Bo; Ren, Juan; Liu, Yue; Huang, Huarong; Zheng, Xi; Zou, Qingze

    2017-03-01

    Abnormalities of blood cholesterol concentration are associated with increased risks for vascular disease, especially heart attacks and strokes. As one of the main lipid components of plasma membrane in all mammalian cells, cholesterol has a major impact on the mechanical properties of the membrane of endothelial cells. Although the effect of cholesterol depletion on cell mechanical properties has been studied, no results yet have been reported on quantitative investigation of cholesterol repletion effect. In this study, the cholesterol repletion effect on the nanomechanical properties of human umbilical vein endothelial cell (EA.hy926) was studied using a control-based atomic force microscope (AFM) nanomechanical measurement protocol. The viscoelasticity of EA.hy926 cells were measured over a large frequency range (0.1-100 Hz) using both constant-rate excitation force with different loading rates and a broadband excitation force. The viscoelasticity oscillation of the cell membranes under the cholesterol effect was also monitored in real-time. The experiment results showed that under the effect of cholesterol repletion, both the Young's modulus and the complex modulus of EA.hy926 cell were increased over 30%, respectively, and moreover, the amplitudes of both the elasticity oscillation and the viscosity oscillation at a period of around 200 s were increased over 70%, respectively. Therefore, this work is among the first to investigate the mechanical properties, particularly, the broadband viscoelasticity variations of EA.hy926 cells under cholesterol repletion treatment. The results revealed that cholesterol repletion may reinforce the coupling of F-actin to plasma membrane by increasing actin stability, and the cholesterol might have modified the submembrane cytoskeletal organization of EA.hy926 cell by causing the involvement of the motor protein nonmuscle myosin II.

  7. Rapid simultaneous determination of organophosphorus pesticides in human serum and urine by liquid chromatography-mass spectrometry

    Directory of Open Access Journals (Sweden)

    Zlatković Milica

    2010-01-01

    Full Text Available Background/Aim. Analysis of organophosphosphorus compounds and their metabolites in a biological material includes the use of numerous methods, covering both preparation of samples for analysis and their identification that is considered to be very complex. Low concentrations monitoring requires implementation of highly sensitive analytical techniques. The aim of this study was to develop and validate an original and sensitive method for the detection and quantitation of organophosphorus pesticides (dimethoate, diazinon, malathion and malaoxon in human biological matrices (serum, urine. Methods. This method was based on a solid-phase extraction procedure, a chromatographic separation using an ACQUITY UPLC ® HSST3 column and mass spectrometric detection in the positve ion mode. Mobile phase: was consited of Solvent A (5 mM ammonium formate pH 3.0 and Solvent B (0.1% acetic formate in methanol, in a linear gradient (constant flow-rate 0.3 mL/min. Results. The standard curve was linear in the range of 0.05-5.00 mg/L for malathion and malaoxon, 0.10-5.00 mg/L for dimethoate and 0.05-2.50 mg/L for diazinon. The correlation coefficient was r ≥ 0.99. Extraction recoveries were satisfactory and ranged between 90-99%. The limits of detection (LOD was between 0.007- 0.07 mg/L and the limits of quantitation (LOQ ranged between 0.022-0.085 mg/L. Intra- and interassay precision and accuracy were satisfactory for all of the pesticides analyzed. Conclusion. The method of liquid chromatography - mass spectrometry is simple, accurate, and useful for the determination of organophosphorus pesticides in both clinical and forensic toxicology.

  8. Rapid determination of lipophilic vitamins in human serum by ultra-high performance liquid chromatography using a fluorinated column and high-throughput miniaturized liquid-liquid extraction.

    Science.gov (United States)

    Cervinkova, Barbora; Krcmova, Lenka Kujovska; Klabackova, Sava; Solichova, Dagmar; Solich, Petr

    2017-09-01

    A high-throughput miniaturized liquid-liquid extraction procedure followed by a simple ultra-high performance liquid chromatography method coupled with fluorescence detection for bioanalytical analysis of all tocopherol isomers and retinol in human serum has been developed and validated. In the extraction procedure, a synthetic internal standard tocol was used, which does not occur in the human body. The separation of structurally related vitamins was achieved using a new generation of pentafluorophenyl propyl core-shell stationary phase with elution using methanol and an aqueous solution of ammonium acetate. The fluorescence of retinol and tocopherol isomers was detected at λex  = 325, 295 nm and λem  = 480, 325 nm, respectively. The rapid baseline separation of all analytes was accomplished within 4.0 min. The sensitivity of method was demonstrated with lower limits of quantification: retinol 0.01 μM, α-tocopherol 0.38 μM, β-tocopherol 0.18 μM, γ-tocopherol 0.14 μM, and δ-tocopherol 0.01 μM. Possible application of this method in clinical practice was confirmed by the analysis of human serum samples from healthy volunteers. Finally, the simultaneous determination of retinol and all tocopherol isomers in human serum can enable the clarification of their role in metabolism and in diseases such as cancer. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Biostability, durability and calcification of cryopreserved human pericardium after rapid glutaraldehyde-stabilization versus multistep ADAPT(R) treatment in a subcutaneous rat model.

    Science.gov (United States)

    Neethling, William; Brizard, Christian; Firth, Laura; Glancy, Ross

    2014-04-01

    Autologous pericardium rapidly fixed with glutaraldehyde (GA) in theatre is considered in many cardiac surgery centres the best material currently available for intracardiac, valvular or vascular repair. Implanted non-fixed autologous tissues suffer rapid degeneration, shrinkage and absorption whereas standard xenotypic fixed tissues cause local cytotoxicity and calcification. In the present study, using a subcutaneous rat model, we tested the biostability, durability and calcification potential of four different pericardium patches treated with GA and relevant to current clinical practice. Pericardium samples were divided into four groups according to the method of treatment. Group I consisted of bovine pericardium (BP) fixed with 0.6% GA (control), Group II cryopreserved human pericardium (CHP) rapidly fixed with 0.6% GA for 4 min and detoxified with MgCl2, Group III CHP treated with the multistep ADAPT(®) process (delipidized, decellularized with Tx-100, deoxycholate, IgePal CA-630 and denucleased, fixed in 0.05% monomeric GA and detoxified) and Group IV BP treated with the multistep ADAPT(®) process (CardioCel(®)). Biostability was determined by shrinkage temperature which measures the degree of cross-linking, and durability assessed by resistance to a mixture of proteinases (pronase digestion). Treated pericardium samples (n = 10 in each of Groups I-IV) were implanted in the subcutaneous rat model for 8 and 16 weeks, followed by histology and calcium analysis (atomic absorption spectrophotometry). The biostability and the durability of both CHP and BP after the multistep ADAPT(®) treatment remained stable without any microscopic calcification. Extractable calcium levels of CHP were significantly (P < 0.01) reduced in Group II (1.89 ± 0.77 μg Ca/mg tissue) compared with Group I (64.37 ± 6.25 μg/mg) after 8 weeks. Calcification of CHP (Group III) and BP (Group IV) after the multistep ADAPT(®) treatment was significantly reduced (1.43 ± 0.48 µg/mg and

  10. Rapid detection of human and canine visceral leishmaniasis: assessment of a latex agglutination test based on the A2 antigen from amastigote forms of Leishmania infantum.

    Science.gov (United States)

    Akhoundi, Behnaz; Mohebali, Mehdi; Shojaee, Saeedeh; Jalali, Mahmoud; Kazemi, Bahram; Bandehpour, Mojgan; Keshavarz, Hossein; Edrissian, Gholam Hossein; Eslami, Mohammad Bagher; Malekafzali, Hossein; Kouchaki, Ameneh

    2013-03-01

    The diagnosis of visceral leishmaniasis (VL) in humans and animal reservoir hosts is difficult, particularly in rural areas where the disease is endemic and laboratory facilities are limited. This study aimed to develop a latex agglutination test (LAT) for the rapid detection of anti-Leishmania antibodies against the A2 antigen derived from the amastigote form as well as those against crude antigens derived from the promastigote form of an Iranian strain of Leishmania (Leishmania) infantum. The A2 antigen (42-100 kDa) was prepared from the amastigote form of L. infantum, purified with electroelution and compared with the crude antigen from the promastigote form of L. infantum. Both antigens showed appropriate intensity reactions, were selected using dot blotting of positive and negative pooled sera and used to sensitize 0.9-μm latex beads. The tests were carried out on sera from 43 symptomatic, human patients with VL confirmed by parasitological examination and direct agglutination test (DAT), 30 healthy controls and 32 patients with other infections but without VL. Canine sera were collected from 63 domestic dogs with VL confirmed using parasitological examinations and DAT and 31 healthy dogs from areas non-endemic for VL. Compared with the controls, human sera from DAT-confirmed patients yielded a sensitivity of 88.4% (95% CI, 82.1-94.5%) and specificity of 93.5% (95% CI, 87.0-99.7%) on A2-LAT (amastigote) when 1:3200 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.914). LAT required 3-5 min to complete, versus the 12-18 h needed for DAT. Compared with the controls, A2-LAT of canine sera from DAT-confirmed cases yielded a sensitivity of 95.2% (95% CI, 95.0-95.4%) and specificity of 100% (95% CI 100%) when 1:320 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.968). Similarly, the sensitivity and specificity of Pro.-LAT (promastigote) was calculated to be 88.4% and 91

  11. Shape memory actuated sliding mechanism for rapid switching between button-patterns for adapted human-machine interaction at different automatization levels in automobiles

    Directory of Open Access Journals (Sweden)

    Sivaperuman Kalairaj Manivannan

    2017-01-01

    Full Text Available Almost all electronics and mechanical equipment have controllers to control the entire or certain tasks. These controllers respond to various types of inputs and performs the function accordingly. One type of input given to the controller is the touch and these controllers which use human touch has touch screen to act as the medium. The touch screen does not give haptic feedback to the users whereas the buttons normally take up more space and cause disturbance and confusion in recognizing the state and the function of the buttons. In this paper, we propose a simple sliding mechanism for rapid switching between different button patterns, in which the buttons remain flat in the surface when inactive and protrude out of the surface when active. The design and simulation are carried out on SolidWorks and the prototypes are fabricated and tested. This mechanism incorporated in the buttons and installed in an automobile, increases the level of automation and enhances the human-machine interaction.

  12. Human young children as well as adults demonstrate 'superior' rapid snake detection when typical striking posture is displayed by the snake.

    Directory of Open Access Journals (Sweden)

    Nobuo Masataka

    2010-11-01

    Full Text Available Humans as well as some nonhuman primates have an evolved predisposition to associate snakes with fear by detecting their presence as fear-relevant stimuli more rapidly than fear-irrelevant ones. In the present experiment, a total of 74 of 3- to 4-year-old children and adults were asked to find a single target black-and-white photo of a snake among an array of eight black-and-white photos of flowers as distracters. As target stimuli, we prepared two groups of snake photos, one in which a typical striking posture was displayed by a snake and the other in which a resting snake was shown. When reaction time to find the snake photo was compared between these two types of the stimuli, its mean value was found to be significantly smaller for the photos of snakes displaying striking posture than for the photos of resting snakes in both the adults and children. These findings suggest the possibility that the human perceptual bias for snakes per se could be differentiated according to the difference of the degree to which their presence acts as a fear-relevant stimulus.

  13. Mach-Zehnder interferometer (MZI) point-of-care system for rapid multiplexed detection of microRNAs in human urine specimens.

    Science.gov (United States)

    Liu, Qing; Shin, Yong; Kee, Jack Sheng; Kim, Kyoung Woo; Mohamed Rafei, Siti Rafeah; Perera, Agampodi Promoda; Tu, Xiaoguang; Lo, Guo-Qiang; Ricci, Estelle; Colombel, Marc; Chiong, Edmund; Thiery, Jean Paul; Park, Mi Kyoung

    2015-09-15

    MicroRNAs have been identified as promising biomarkers for human diseases. The development of a point-of-care (POC) test for the disease-associated miRNAs would be especially beneficial, since miRNAs are unexpectedly well preserved in various human specimens, including urine. Here, we present the Mach-Zehnder interferometer-miRNA detection system capable of detecting multiple miRNAs in clinical urine samples rapidly and simultaneously in a label-free and real-time manner. Through measurement of the light phase change, the MZI sensor provides an optical platform for fast profiling of small molecules with improved accuracy. We demonstrate that this system could specifically detect target miRNAs (miR-21, and let-7a), and even identify the single nucleotide polymorphism of the let-7 family of miRNAs from synthetic and cell line samples. The clinical applicability of this system is confirmed by simultaneously detecting two types of miRNAs in urine samples of bladder cancer patients in a single reaction, with a detection time of 15 min. The POC system can be expanded to detect a number of miRNAs of different species and should be useful for a variety of clinical applications requiring at or near the site of patient care. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Rapid and simultaneous determination of essential minerals and trace elements in human milk by improved flame atomic absorption spectroscopy (FAAS) with microwave digestion.

    Science.gov (United States)

    Luo, Yang; Zhang, Bo; Chen, Ming; Wang, Jue; Zhang, Xue; Gao, Wei-Yin; Huang, Jun-Fu; Fu, Wei-Ling

    2010-09-08

    A method for the simultaneous and economical determination of many trace elements in human milk is developed. Two multi-element hollow cathode lamps (HCLs) were used instead of single-element HCLs to improve the sample throughput of flame atomic absorption spectroscopy (FAAS). The microwave digestion of milk is optimized prior to detection, and the performance characteristics of the improved analysis method are identified. Clinical samples are detected by both FAAS and inductively coupled plasma-optical emission spectroscopy (ICP-OES) for methodology evaluation. Results reveal that the proposed FAAS with multi-element HCLs could determine six essential minerals and trace elements within 15 min. This method provides a linear analytical range of 0.01-10 mg L(-1). For Ca, Cu, Fe, Mg, Mn, and Zn, the limits of determination are 1.5, 3, 1.8, 2.2, 2.1, and 1.3 microg L(-1), respectively. The mean relative standard deviations (RSDs) of intra- and interassays are lower than 7%. Excellent operational characteristics of rapidity, simplicity, and economy make the proposed method a promising one for the quantification of trace elements in human milk in clinics of underdeveloped areas.

  15. Growing Concerns With Workplace Incivility.

    Science.gov (United States)

    Collins, Natasha Renee; Rogers, Bonnie

    2017-11-01

    Workplace incivility (WPI) is a growing issue across all public and private sectors. Occupational and environmental health nurses can educate employees and management about WPI, its risk factors and characteristics, and ways to reduce incidents of WPI.

  16. Validation of a novel and rapid method for the simultaneous determination of some phenolic organohalogens in human serum by GC-MS.

    Science.gov (United States)

    Dufour, Patrice; Pirard, Catherine; Charlier, Corinne

    2016-11-15

    Over the last decades, more and more studies focused on the impact of endocrine disruptors on the environment and human health. Among them, phenolic organohalogens (POHs) are a particular concern because of their structural resemblance with natural hormones. There are different methods that are known to quantify these compounds in human serum, however, the current extraction techniques are long, fastidious and using harmfull chemicals such as diazomethane and sulfuric acid. Consequently, we developed an alternative, sensitive and faster method to simultaneously quantify pentachlorophenol (PCP), tetrabromobisphenol A (TBBPA), 4 bromophenols, 7 hydroxypolychlorinated biphenyls (OH-PCBs) and 3 hydroxy-polybrominated diphenyl ether (OH-PBDEs) in human serum sample. The clean-up and the enrichment of the sample were performed in a single extraction step using strong anion-exchange solid phase cartridge. After a rapid liquid-liquid extraction step to remove acidic traces, the extract was derivatized using trimethylsilyldiazomethane (TMSD) and finally analyzed by a gas-chromatograph coupled with an electron negative capture chemical ionization source combined with a triple quadrupole mass spectrometer (GC-ENCI-MS) operating in single ion monitoring. The whole procedure was validated according to the total error approach. The inter and intra assay precision were demonstrated to be lower than 20% and the relative bias to be lower than 15% in the dosing range of concentrations. The limit of quantification (LOQ) ranged from 2pgmL-1 and 5pgmL-1, except for the PCP (44.6pgmL-1) and for the 2,4,6-tribromophenol (49.6pgmL-1). Finally, the method was successfully applied to measure the POH background contamination in serum samples collected from 20 Belgian blood donors recruited in CHU Mont-Godinne (Namur, Belgium) aged between 21 and 69 years old. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Recombination dynamics of a human Y-chromosomal palindrome: rapid GC-biased gene conversion, multi-kilobase conversion tracts, and rare inversions.

    Science.gov (United States)

    Hallast, Pille; Balaresque, Patricia; Bowden, Georgina R; Ballereau, Stéphane; Jobling, Mark A

    2013-01-01

    The male-specific region of the human Y chromosome (MSY) includes eight large inverted repeats (palindromes) in which arm-to-arm similarity exceeds 99.9%, due to gene conversion activity. Here, we studied one of these palindromes, P6, in order to illuminate the dynamics of the gene conversion process. We genotyped ten paralogous sequence variants (PSVs) within the arms of P6 in 378 Y chromosomes whose evolutionary relationships within the SNP-defined Y phylogeny are known. This allowed the identification of 146 historical gene conversion events involving individual PSVs, occurring at a rate of 2.9-8.4×10(-4) events per generation. A consideration of the nature of nucleotide change and the ancestral state of each PSV showed that the conversion process was significantly biased towards the fixation of G or C nucleotides (GC-biased), and also towards the ancestral state. Determination of haplotypes by long-PCR allowed likely co-conversion of PSVs to be identified, and suggested that conversion tract lengths are large, with a mean of 2068 bp, and a maximum in excess of 9 kb. Despite the frequent formation of recombination intermediates implied by the rapid observed gene conversion activity, resolution via crossover is rare: only three inversions within P6 were detected in the sample. An analysis of chimpanzee and gorilla P6 orthologs showed that the ancestral state bias has existed in all three species, and comparison of human and chimpanzee sequences with the gorilla outgroup confirmed that GC bias of the conversion process has apparently been active in both the human and chimpanzee lineages.

  18. Application of host-specific source-tracking tools for rapid identification of fecal contamination in fresh produce by humans and livestock.

    Science.gov (United States)

    Lee, Chang Soo; Lee, Jiyoung

    2013-03-30

    Fecal contamination in fresh produce is a public health concern because it may contain human pathogens. We introduced host-specific quantitative real-time polymerase chain reaction (qPCR) assays for the rapid detection and identification of fecal contamination sources from humans and farm animals (cow, pig, chicken) in fresh produce. Each composite fecal sample was spiked on lettuce at two contamination levels (0.2 mg or 2 mg feces g⁻¹), followed by qPCR assays for detecting each host-specific genetic marker: BoBac (cow); PF163 (pig); CP3-49 (chicken); and HF183 and gyrB (human). Two commercial DNA extraction kits were compared to evaluate DNA recovery yields and removal of PCR inhibition. Sketa2 assay was conducted to assess the presence of PCR inhibition in the contaminated lettuce. All the qPCR assays yielded reliable detection from contaminated lettuce (2 mg feces g⁻¹), where their target gene numbers were 1.5-5.0 × 10³ (HF183), 0.8-2.2 × 10³ (gyrB), 0.6-1.6 × 10³ (BoBac), 1.6-3.0 × 10³ (CP3-49) and 1.1-2.2 × 10³ (PF163) copies g⁻¹ of lettuce. Among the two extraction kits, QIAamp DNA Stool Kit resulted in 2-3 times higher sensitivity and 20% less PCR inhibition than the PowerFood™ kit. This study provides information on the optimized host-specific qPCR assay in identifying sources of fecal contamination in fresh produce and is useful for tracking the contamination source and improving agricultural practice. © 2012 Society of Chemical Industry.

  19. Recombination dynamics of a human Y-chromosomal palindrome: rapid GC-biased gene conversion, multi-kilobase conversion tracts, and rare inversions.

    Directory of Open Access Journals (Sweden)

    Pille Hallast

    Full Text Available The male-specific region of the human Y chromosome (MSY includes eight large inverted repeats (palindromes in which arm-to-arm similarity exceeds 99.9%, due to gene conversion activity. Here, we studied one of these palindromes, P6, in order to illuminate the dynamics of the gene conversion process. We genotyped ten paralogous sequence variants (PSVs within the arms of P6 in 378 Y chromosomes whose evolutionary relationships within the SNP-defined Y phylogeny are known. This allowed the identification of 146 historical gene conversion events involving individual PSVs, occurring at a rate of 2.9-8.4×10(-4 events per generation. A consideration of the nature of nucleotide change and the ancestral state of each PSV showed that the conversion process was significantly biased towards the fixation of G or C nucleotides (GC-biased, and also towards the ancestral state. Determination of haplotypes by long-PCR allowed likely co-conversion of PSVs to be identified, and suggested that conversion tract lengths are large, with a mean of 2068 bp, and a maximum in excess of 9 kb. Despite the frequent formation of recombination intermediates implied by the rapid observed gene conversion activity, resolution via crossover is rare: only three inversions within P6 were detected in the sample. An analysis of chimpanzee and gorilla P6 orthologs showed that the ancestral state bias has existed in all three species, and comparison of human and chimpanzee sequences with the gorilla outgroup confirmed that GC bias of the conversion process has apparently been active in both the human and chimpanzee lineages.

  20. Rapid and simultaneous determination of five vinca alkaloids in Catharanthus roseus and human serum using trilinear component modeling of liquid chromatography-diode array detection data.

    Science.gov (United States)

    Liu, Zhi; Wu, Hai-Long; Li, Yong; Gu, Hui-Wen; Yin, Xiao-Li; Xie, Li-Xia; Yu, Ru-Qin

    2016-07-15

    A novel chemometrics-assisted high performance liquid chromatography method coupled with diode array detector (HPLC-DAD) was proposed for the simultaneous determination of vincristine (VCR), vinblastine (VLB), vindoline (VDL), catharanthine (CAT) and yohimbine (YHB) in Catharanthus roseus (C. roseus) and human serum samples. With the second-order advantage of the alternating trilinear decomposition (ATLD) method, the resolution and rapid determination of five components of interest in complex matrices were performed, even in the present of heavy overlaps and unknown interferences. Therefore, multi-step purification was omitted and five components could be fast eluted out within 7.5min under simple isocratic elution condition (acetonitrile/0.2% formic acid water, 37:63, v/v). Statistical parameters, such as the linear correlation coefficient (R(2)), root-mean-square error of prediction (RMSEP), limit of detection (LOD) and limit of quantitation (LOQ) had been calculated to investigate the accuracy and reliability of the method. The average recoveries of five vinca alkaloids ranged from 97.1% to 101.9% and 98.8% to 103.0% in C. roseus and human serum samples, respectively. The five vinca alkaloids were adequately determined with limits of detection (LODs) of 29.5-49.3ngmL(-1) in C. roseus and 12.4-27.2ngmL(-1) in human serum samples, respectively. The obtained results demonstrated that the analytical strategy provided a feasible alternative for synchronously monitoring the quality of raw herb and the concentration of blood drugs. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Nanomaterial-enabled Rapid Detection of Water Contaminants.

    Science.gov (United States)

    Mao, Shun; Chang, Jingbo; Zhou, Guihua; Chen, Junhong

    2015-10-28

    Water contaminants, e.g., inorganic chemicals and microorganisms, are critical metrics for water quality monitoring and have significant impacts on human health and plants/organisms living in water. The scope and focus of this review is nanomaterial-based optical, electronic, and electrochemical sensors for rapid detection of water contaminants, e.g., heavy metals, anions, and bacteria. These contaminants are commonly found in different water systems. The importance of water quality monitoring and control demands significant advancement in the detection of contaminants in water because current sensing technologies for water contaminants have limitations. The advantages of nanomaterial-based sensing technologies are highlighted and recent progress on nanomaterial-based sensors for rapid water contaminant detection is discussed. An outlook for future research into this rapidly growing field is also provided. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum

    Science.gov (United States)

    Krastins, Bryan; Prakash, Amol; Sarracino, David A.; Nedelkov, Dobrin; Niederkofler, Eric E.; Kiernan, Urban A.; Nelson, Randall; Vogelsang, Maryann S.; Vadali, Gouri; Garces, Alejandra; Sutton, Jennifer N.; Peterman, Scott; Byram, Gregory; Darbouret, Bruno; Pérusse, Joëlle R.; Seidah, Nabil G.; Coulombe, Benoit; Gobom, Johan; Portelius, Erik; Pannee, Josef; Blennow, Kaj; Kulasingam, Vathany; Couchman, Lewis; Moniz, Caje; Lopez, Mary F.

    2013-01-01

    Objectives The aim of this study was to develop high-throughput, quantitative and highly selective mass spectrometric, targeted immunoassays for clinically important proteins in human plasma or serum. Design and methods The described method coupled mass spectrometric immunoassay (MSIA), a previously developed technique for immunoenrichment on a monolithic microcolumn activated with an anti-protein antibody and fixed in a pipette tip, to selected reaction monitoring (SRM) detection and accurate quantification of targeted peptides, including clinically relevant sequence or truncated variants. Results In this report, we demonstrate the rapid development of MSIA-SRM assays for sixteen different target proteins spanning seven different clinically important areas (including neurological, Alzheimer's, cardiovascular, endocrine function, cancer and other diseases) and ranging in concentration from pg/mL to mg/mL. The reported MSIA-SRM assays demonstrated high sensitivity (within published clinical ranges), precision, robustness and high-throughput as well as specific detection of clinically relevant isoforms for many of the target proteins. Most of the assays were tested with bona-fide clinical samples. In addition, positive correlations, (R2 0.67–0.87, depending on the target peptide), were demonstrated for MSIA-SRM assay data with clinical analyzer measurements of parathyroid hormone (PTH) and insulin growth factor 1 (IGF1) in clinical sample cohorts. Conclusions We have presented a practical and scalable method for rapid development and deployment of MS-based SRM assays for clinically relevant proteins and measured levels of the target analytes in bona fide clinical samples. The method permits the specific quantification of individual protein isoforms and addresses the difficult problem of protein heterogeneity in clinical proteomics applications. PMID:23313081

  3. Importance of a Rapid and Accurate Diagnosis inStrongyloides Stercoralisand Human T-Lymphotropic Virus 1 Co-infection: A Case Report and Review of the Literature.

    Science.gov (United States)

    Quintero, Olga; Berini, Carolina A; Waldbaum, Carlos; Avagnina, Alejandra; Juarez, María; Repetto, Silvia; Sorda, Juan; Biglione, Mirna

    2017-01-01

    Strongyloides (S.) stercoralis and Human T-Lymphotropic Virus 1 (HTLV-1) share some endemic regions such as Japan, Jamaica, and South America and are mostly diagnosed elsewhere in immigrants from endemic areas. This co-infection has not been documented in Argentina although both pathogens are endemic in the Northwest. We present a case of S. stercoralis and HTLV-1 co-infection with an initial presentation due to gastrointestinal symptoms which presented neither eosinophilia nor the presence of larvae in stool samples in a non-endemic area for these infections. A young Peruvian woman living in Buenos Aires attended several emergency rooms and finally ended up admitted in a gastroenterology ward due to incoercible vomiting, diarrhea, abdominal pain, fever, and weight loss. Gastrointestinal symptoms started 3 months before she returned to Argentina from a trip to Peru. She presented malnutrition and abdominal distension parameters. HIV-1 and other immunodeficiencies were discarded. The serial coproparasitological test was negative. Computed tomography showed diffuse thickening of duodenal and jejunal walls. At the beginning, vasculitis was suspected and corticosteroid therapy was initiated. The patient worsened rapidly. Skin, new enteral biopsies, and a new set of coproparasitological samples revealed S. stercoralis . Then, HTLV-1 was suspected and infection was confirmed. Ivermectin and albendazole were administrated, until the stool sample remained negative for 2 weeks. Larvae were not observed in fresh stool, Ritchie method, and agar culture 1 week post-treatment. Although she required initial support with parenteral nutrition due to oral intolerance she slowly progressed favorably. It has been highly recommended to include a rapid and sensitive PCR strategy in the algorithm to confirm Strongyloides infection, which has demonstrated to improve early diagnosis in patients at-risk of disseminated strongyloidiasis.

  4. Growing Oppression, Growing Resistance : LGBT Activism and Europeanisation in Macedonia

    NARCIS (Netherlands)

    Miškovska Kajevska, A.; Bilić, B.

    2016-01-01

    This chapter provides one of the first socio-historical overviews of the LGBT groups in Macedonia and argues that an important impetus for the proliferation of LGBT activities has been the growing state-endorsed homophobia starting from 2008. The homophobic rhetoric of the ruling parties was clearly

  5. Human CD4+ CD25hi Foxp3+ regulatory T cells are derived by rapid turnover of memory populations in vivo

    Science.gov (United States)

    Vukmanovic-Stejic, Milica; Zhang, Yan; Cook, Joanne E.; Fletcher, Jean M.; McQuaid, Arthur; Masters, Joanne E.; Rustin, Malcolm H.A.; Taams, Leonie S.; Beverley, Peter C.L.; Macallan, Derek C.; Akbar, Arne N.

    2006-01-01

    While memory T cells are maintained by continuous turnover, it is not clear how human regulatory CD4+CD45RO+CD25hi Foxp3+ T lymphocyte populations persist throughout life. We therefore used deuterium labeling of cycling cells in vivo to determine whether these cells could be replenished by proliferation. We found that CD4+CD45RO+Foxp3+CD25hi T lymphocytes were highly proliferative, with a doubling time of 8 days, compared with memory CD4+CD45RO+Foxp3–CD25– (24 days) or naive CD4+CD45RA+Foxp3–CD25– populations (199 days). However, the regulatory population was susceptible to apoptosis and had critically short telomeres and low telomerase activity. It was therefore unlikely to be self regenerating. These data are consistent with continuous production from another population source. We found extremely close TCR clonal homology between regulatory and memory CD4+ T cells. Furthermore, antigen-related expansions within certain TCR Vβ families were associated with parallel numerical increases of CD4+CD45RO+CD25hiFoxp3+ Tregs with the same Vβ usage. It is therefore unlikely that all human CD4+CD25+Foxp3+ Tregs are generated as a separate functional lineage in the thymus. Instead, our data suggest that a proportion of this regulatory population is generated from rapidly dividing, highly differentiated memory CD4+ T cells; this has considerable implications for the therapeutic manipulation of these cells in vivo. PMID:16955142

  6. Rapid screening and multi-toxin profile confirmation of tetrodotoxins and analogues in human body fluids derived from a puffer fish poisoning incident in New Caledonia.

    Science.gov (United States)

    Rambla-Alegre, Maria; Leonardo, Sandra; Barguil, Yann; Flores, Cintia; Caixach, Josep; Campbell, Katrina; Elliott, Christopher T; Maillaud, Claude; Boundy, Michael J; Harwood, D Tim; Campàs, Mònica; Diogène, Jorge

    2018-02-01

    In August 2014, a puffer fish poisoning incidence resulting in one fatality was reported in New Caledonia. Although tetrodotoxin (TTX) intoxication was established from the patients' signs and symptoms, the determination of TTX in the patient's urine, serum or plasma is essential to confirm the clinical diagnosis. To provide a simple cost-effective rapid screening tool for clinical analysis, a maleimide-based enzyme-linked immunosorbent assay (mELISA) adapted for the determination of TTX contents in human body fluids was assessed. The mELISA was applied to the analysis of urine samples from two patients and a response for the presence of TTX and/or structurally similar analogues was detected in all samples. The analysis by LC-MS/MS confirmed the presence of TTX but also TTX analogues (4-epiTTX, 4,9-anhydroTTX and 5,6,11-trideoxyTTX) in the urine. A change in the multi-toxin profile in the urine based on time following consumption was observed. LC-MS/MS analysis of serum and plasma samples also revealed the presence of TTX (32.9 ng/mL) and 5,6,11-trideoxyTTX (374.6 ng/mL) in the post-mortem plasma. The results provide for the first time the TTX multi-toxin profile of human samples from a puffer fish intoxication and clearly demonstrate the implication of TTX as the causative agent of the reported intoxication case. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Human kallikrein 2 (hK2), but not prostate-specific antigen (PSA), rapidly complexes with protease inhibitor 6 (PI-6) released from prostate carcinoma cells.

    Science.gov (United States)

    Saedi, M S; Zhu, Z; Marker, K; Liu, R S; Carpenter, P M; Rittenhouse, H; Mikolajczyk, S D

    2001-11-01

    Human kallikrein 2 (hK2) is a secreted, trypsin-like protease that shares 80% amino acid sequence identity with prostate-specific antigen (PSA). hK2 has been shown to be a serum marker for prostate cancer and may also play a role in cancer progression and metastasis. We have previously identified a novel complex between human kallikrein 2 (hK2) and protease inhibitor 6 (PI-6) in prostate cancer tissue. PI-6 is an intracellular serine protease inhibitor with both antitrypsin and antichymotrypsin activity. In the current study we have shown that PI-6 forms a rapid in vitro complex with hK2 but does not complex with PSA. Recombinant mammalian cells expressing both hK2 and PI-6 showed hK2-PI-6 complex in the spent media only after cell death and lysis. Similarly, LNCaP cells expressing endogenous hK2 and PI-6 showed extracellular hK2-PI-6 complex formation concurrently with cell death. Immunostaining of prostate cancer tissues with PI-6 monoclonal antibodies showed a marked preferential staining pattern in cancerous epithelial cells compared with noncancerous tissue. These results indicate that the hK2-PI-6 complex may be a naturally occurring marker of tissue damage and necrosis associated with neoplasia. Both hK2 and PI-6 were shed into the lumen of prostate cancer glands as granular material that appeared to be cellular necrotic debris. The differential staining pattern of PI6 in tissues suggests a complex regulation of PI-6 expression that may play a role in other aspects of neoplastic progression. Copyright 2001 Wiley-Liss, Inc.

  8. Threats to global health and survival: the growing crises of tropical infectious diseases--our "unfinished agenda".

    Science.gov (United States)

    Guerrant, R L; Blackwood, B L

    1999-05-01

    Health, one of our most unassailable human values, transcends all geographic, political, and cultural boundaries. The health problems of the rapidly growing 80% of the world's population that live in the tropical developing countries of Asia, Africa, and Latin America pose major threats to industrialized as well as developing regions. These threats can be divided into three areas, or three "E"s: (1) emerging, reemerging, and antimicrobial-resistant infections; (2) exploding populations without improved health; and (3) erosion of our humanity or leadership if we ignore the growing health problems of the poor. Our assessment of current trends in global population distribution and resource consumption; DALY calculations, causes, and distribution of global mortality and morbidity; and the misperceptions about and maldistribution of resources for health point to the critical importance of addressing tropical infectious diseases and global health for preservation of democracy and civilization as we know it.

  9. Large volume sample stacking for rapid and sensitive determination of antidiabetic drug metformin in human urine and serum by capillary electrophoresis with contactless conductivity detection.

    Science.gov (United States)

    Tůma, Petr

    2014-06-06

    Two CE methods with contactless conductivity detection have been developed for determining the oral antidiabetic drug metformin in human urine and blood. The determination of metformin is performed on a separation capillary with an effective length of 14 cm, using a maximum voltage of 30 kV and with a small injection of 50-fold diluted urine into the capillary. Under these conditions, the migration time of metformin is 35s and the LOD is 0.3 μM. Large-volume sample stacking was used to determine low metformin levels in serum. The injection of a sample of serum deproteinized with acetonitrile was 10 times greater compared to the injected amount of urine. This enabled reduction of the LOD to 0.03 μM and the metformin migration time equalled 86 s. The undesirable solvent from sample zone was forced out of the capillary to ensure rapidity and good repeatability of the determination. The RSD values for the migration time are 0.1% for urine and 0.7% for serum; RSD for the peak areas equalled 1.4% for urine and 2.6% for serum. The developed CE technique was tested on performance of routine analyses of metformin in the urine and serum of patients suffering from type II diabetes mellitus. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Stunning and Right Ventricular Dysfunction Is Induced by Coronary Balloon Occlusion and Rapid Pacing in Humans: Insights From Right Ventricular Conductance Catheter Studies.

    Science.gov (United States)

    Axell, Richard G; Giblett, Joel P; White, Paul A; Klein, Andrew; Hampton-Til, James; O'Sullivan, Michael; Braganza, Denise; Davies, William R; West, Nick E J; Densem, Cameron G; Hoole, Stephen P

    2017-06-06

    We sought to determine whether right ventricular stunning could be detected after supply (during coronary balloon occlusion [BO]) and supply/demand ischemia (induced by rapid pacing [RP] during transcatheter aortic valve replacement) in humans. Ten subjects with single-vessel right coronary artery disease undergoing percutaneous coronary intervention with normal ventricular function were studied in the BO group. Ten subjects undergoing transfemoral transcatheter aortic valve replacement were studied in the RP group. In both, a conductance catheter was placed into the right ventricle, and pressure volume loops were recorded at baseline and for intervals over 15 minutes after a low-pressure BO for 1 minute or a cumulative duration of RP for up to 1 minute. Ischemia-induced diastolic dysfunction was seen 1 minute after RP (end-diastolic pressure [mm Hg]: 8.1±4.2 versus 12.1±4.1, P right coronary artery balloon occlusion both cause ischemic right ventricular dysfunction with stunning observed later during the procedure. This may have intraoperative implications in patients without right ventricular functional reserve. © 2017 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

  11. Coffee consumption rapidly reduces background DNA strand breaks in healthy humans: Results of a short-term repeated uptake intervention study.

    Science.gov (United States)

    Bakuradze, Tamara; Lang, Roman; Hofmann, Thomas; Schipp, Dorothea; Galan, Jens; Eisenbrand, Gerhard; Richling, Elke

    2016-03-01

    Intervention studies provide evidence that long-term coffee consumption correlates with reduced DNA background damage in healthy volunteers. Here, we report on short-term kinetics of this effect, showing a rapid onset after normal coffee intake. In a short-term human intervention study, we determined the effects of coffee intake on DNA integrity during 8 h. Healthy male subjects ingested coffee in 200 mL aliquots every second hour up to a total volume of 800 mL. Blood samples were taken at baseline, immediately before the first coffee intake and subsequently every 2 h, prior to the respective coffee intake. DNA integrity was assayed by the comet assay. The results show a significant (p coffee intake. Continued coffee intake was associated with further decrements in background DNA damage within the 8 h intervention (p coffee consumption). Repeated coffee consumption was associated with reduced background DNA strand breakage, clearly measurable as early as 2 h after first intake resulting in a cumulative overall reduction by about one-third of the baseline value. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Rapid determination of three alkaloids from Lotus Plumule in human serum using an HPLC-DAD method with a short monolithic column.

    Science.gov (United States)

    Du, Hui; Ren, Jing; Wang, Sicen

    2011-12-01

    An HPLC-DAD with a short monolithic column method was used for rapid, sensitive and simultaneous determination of liensinine, isoliensinine and neferine in human serum. Chromatographic separation was achieved on an RP-HPLC chromolithic column (50mm×4.6mm i.d.), with a mobile phase of methanol-water-triethylamine-acetic acid (70:30:0.2:0.05), at a flow rate of 1mlmin(-1). The calibration curves of three alkaloids exhibit good linear relationship in the concentration range of 0.5-80μgml(-1). The limits of detection (LODs) of three alkaloids were 20, 20 and 25ngml(-1), respectively, and the separation time was 4min. A comparative study has been performed with a normal C18 packed column (150mm×4.6mm i.d., 5μm). The separation time was 16min and the LODs of three alkaloids were 56, 56 and 65ngml(-1), respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Rapid acquisition of helium-3 and proton three-dimensional image sets of the human lung in a single breath-hold using compressed sensing.

    Science.gov (United States)

    Qing, Kun; Altes, Talissa A; Tustison, Nicholas J; Feng, Xue; Chen, Xiao; Mata, Jaime F; Miller, G Wilson; de Lange, Eduard E; Tobias, William A; Cates, Gordon D; Brookeman, James R; Mugler, John P

    2015-10-01

    To develop and validate a method for acquiring helium-3 ((3) He) and proton ((1) H) three-dimensional (3D) image sets of the human lung with isotropic spatial resolution within a 10-s breath-hold by using compressed sensing (CS) acceleration, and to assess the fidelity of undersampled images compared with fully sampled images. The undersampling scheme for CS acceleration was optimized and tested using (3) He ventilation data. Rapid 3D acquisition of both (3) He and (1) H data during one breath-hold was then implemented, based on a balanced steady-state free-precession pulse sequence, by random undersampling of k-space with reconstruction by means of minimizing the L1 norm and total variance. CS-reconstruction fidelity was evaluated quantitatively by comparing fully sampled and retrospectively undersampled image sets. Helium-3 and (1) H 3D image sets of the lung with isotropic 3.9-mm resolution were acquired during a single breath-hold in 12 s and 8 s using acceleration factors of 2 and 3, respectively. Comparison of fully sampled and retrospectively undersampled (3) He and (1) H images yielded mean absolute errors 0.9. By randomly undersampling k-space and using CS reconstruction, high-quality (3) He and (1) H 3D image sets with isotropic 3.9-mm resolution can be acquired within an 8-s breath-hold. © 2014 Wiley Periodicals, Inc.

  14. The Silk-protein Sericin Induces Rapid Melanization of Cultured Primary Human Retinal Pigment Epithelial Cells by Activating the NF-κB Pathway.

    Science.gov (United States)

    Eidet, J R; Reppe, S; Pasovic, L; Olstad, O K; Lyberg, T; Khan, A Z; Fostad, I G; Chen, D F; Utheim, T P

    2016-03-04

    Restoration of the retinal pigment epithelial (RPE) cells to prevent further loss of vision in patients with age-related macular degeneration represents a promising novel treatment modality. Development of RPE transplants, however, requires up to 3 months of cell differentiation. We explored whether the silk protein sericin can induce maturation of primary human retinal pigment epithelial (hRPE) cells. Microarray analysis demonstrated that sericin up-regulated RPE-associated transcripts (RPE65 and CRALBP). Upstream analysis identified the NF-κB pathway as one of the top sericin-induced regulators. ELISA confirmed that sericin stimulates the main NF-κB pathway. Increased levels of RPE-associated proteins (RPE65 and the pigment melanin) in the sericin-supplemented cultures were confirmed by western blot, spectrophotometry and transmission electron microscopy. Sericin also increased cell density and reduced cell death following serum starvation in culture. Inclusion of NF-κB agonists and antagonists in the culture medium showed that activation of the NF-κB pathway appears to be necessary, but not sufficient, for sericin-induced RPE pigmentation. We conclude that sericin promotes pigmentation of cultured primary hRPE cells by activating the main NF-κB pathway. Sericin's potential role in culture protocols for rapid differentiation of hRPE cells derived from embryonic or induced pluripotent stem cells should be investigated.

  15. Rapid determination of acetone in human blood by derivatization with pentafluorobenzyl hydroxylamine followed by headspace liquid-phase microextraction and gas chromatography/mass spectrometry.

    Science.gov (United States)

    Deng, Chunhui; Li, Ning; Wang, Xiaochuan; Zhang, Xiangmin; Zeng, Jia

    2005-01-01

    In the current work, a simple, rapid, accurate and inexpensive method was developed for the determination of acetone in human blood. The proposed method is based on derivatization with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA), followed by headspace liquid-phase microextraction (HS-LPME) and gas chromatography/mass spectrometry (GC/MS). In the present method, acetone in blood samples was derivatized with PFBHA and acetone oxime formed in several seconds. The formed oxime was enriched by HS-LPME using the organic solvent film (OSF) formed in a microsyringe barrel as extraction interface. Finally, the enriched oxime was analyzed by GC/MS in electron ionization (EI) mode. HS-LPME parameters including solvent, syringe plunger withdrawal rate, sampling volume, and extraction cycle were optimized and the method reproducibility, linearity, recovery and detection limit were studied. The proposed method was applied to determination of acetone in diabetes blood and normal blood. It has been shown that derivatization with HS-LPME and GC/MS is an alternative method for determination of the diabetes biomarker, acetone, in blood samples.

  16. Rapid small lot manufacturing

    Energy Technology Data Exchange (ETDEWEB)

    Harrigan, R.W.

    1998-05-09

    The direct connection of information, captured in forms such as CAD databases, to the factory floor is enabling a revolution in manufacturing. Rapid response to very dynamic market conditions is becoming the norm rather than the exception. In order to provide economical rapid fabrication of small numbers of variable products, one must design with manufacturing constraints in mind. In addition, flexible manufacturing systems must be programmed automatically to reduce the time for product change over in the factory and eliminate human errors. Sensor based machine control is needed to adapt idealized, model based machine programs to uncontrolled variables such as the condition of raw materials and fabrication tolerances.

  17. Exploring Classroom Hydroponics. Growing Ideas.

    Science.gov (United States)

    National Gardening Association, Burlington, VT.

    Growing Ideas, the National Gardening Association's series for elementary, middle, and junior high school educators, helps teachers engage students in using plants and gardens as contexts for developing a deeper, richer understanding of the world around them. This volume's focus is on hydroponics. It presents basic hydroponics information along…

  18. Growing Patterns: Seeing beyond Counting

    Science.gov (United States)

    Markworth, Kimberly A.

    2012-01-01

    Over the past two decades, mathematical patterns have been acknowledged as important early components of children's development of algebraic reasoning (NCTM 2000). In particular, growing patterns have attracted significant attention as a context that helps students develop an understanding of functional relationships (Lee and Freiman 2006; Moss et…

  19. Organization of growing random networks

    Energy Technology Data Exchange (ETDEWEB)

    Krapivsky, P. L.; Redner, S.

    2001-06-01

    The organizational development of growing random networks is investigated. These growing networks are built by adding nodes successively, and linking each to an earlier node of degree k with an attachment probability A{sub k}. When A{sub k} grows more slowly than linearly with k, the number of nodes with k links, N{sub k}(t), decays faster than a power law in k, while for A{sub k} growing faster than linearly in k, a single node emerges which connects to nearly all other nodes. When A{sub k} is asymptotically linear, N{sub k}(t){similar_to}tk{sup {minus}{nu}}, with {nu} dependent on details of the attachment probability, but in the range 2{lt}{nu}{lt}{infinity}. The combined age and degree distribution of nodes shows that old nodes typically have a large degree. There is also a significant correlation in the degrees of neighboring nodes, so that nodes of similar degree are more likely to be connected. The size distributions of the in and out components of the network with respect to a given node{emdash}namely, its {open_quotes}descendants{close_quotes} and {open_quotes}ancestors{close_quotes}{emdash}are also determined. The in component exhibits a robust s{sup {minus}2} power-law tail, where s is the component size. The out component has a typical size of order lnt, and it provides basic insights into the genealogy of the network.

  20. Evaluation of the suitability of a plant virus, pepper mild mottle virus, as a surrogate of human enteric viruses for assessment of the efficacy of coagulation-rapid sand filtration to remove those viruses.

    Science.gov (United States)

    Shirasaki, N; Matsushita, T; Matsui, Y; Yamashita, R

    2018-02-01

    Here, we evaluated the removal of three representative human enteric viruses - adenovirus (AdV) type 40, coxsackievirus (CV) B5, and hepatitis A virus (HAV) IB - and one surrogate of human caliciviruses - murine norovirus (MNV) type 1 - by coagulation-rapid sand filtration, using water samples from eight water sources for drinking water treatment plants in Japan. The removal ratios of a plant virus (pepper mild mottle virus; PMMoV) and two bacteriophages (MS2 and φX174) were compared with the removal ratios of human enteric viruses to assess the suitability of PMMoV, MS2, and φX174 as surrogates for human enteric viruses. The removal ratios of AdV, CV, HAV, and MNV, evaluated via the real-time polymerase chain reaction (PCR) method, were 0.8-2.5-log10 when commercially available polyaluminum chloride (PACl, basicity 1.5) and virgin silica sand were used as the coagulant and filter medium, respectively. The type of coagulant affected the virus removal efficiency, but the age of silica sand used in the rapid sand filtration did not. Coagulation-rapid sand filtration with non-sulfated, high-basicity PACls (basicity 2.1 or 2.5) removed viruses more efficiently than the other aluminum-based coagulants. The removal ratios of MS2 were sometimes higher than those of the three human enteric viruses and MNV, whereas the removal ratios of φX174 tended to be smaller than those of the three human enteric viruses and MNV. In contrast, the removal ratios of PMMoV were similar to and strongly correlated with those of the three human enteric viruses and MNV. Thus, PMMoV appears to be a suitable surrogate for human enteric viruses for the assessment of the efficacy of coagulation-rapid sand filtration to remove viruses. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Rapid Prototyping

    Science.gov (United States)

    1999-01-01

    Javelin, a Lone Peak Engineering Inc. Company has introduced the SteamRoller(TM) System as a commercial product. The system was designed by Javelin during a Phase II NASA funded small commercial product. The purpose of the invention was to allow automated-feed of flexible ceramic tapes to the Laminated Object Manufacturing rapid prototyping equipment. The ceramic material that Javelin was working with during the Phase II project is silicon nitride. This engineered ceramic material is of interest for space-based component.

  2. Development of a lateral flow immunochromatographic assay for rapid detection of Mycoplasma pneumoniae-specific IgM in human serum specimens.

    Science.gov (United States)

    Ou, Liming; Lv, Qingyu; Wu, Canjun; Hao, Huaijie; Zheng, Yuling; Jiang, Yongqiang

    2016-05-01

    Early diagnosis of Mycoplasma pneumoniae (MP) infection is crucial for prompt treatment and good patient outcome. However, serological tests to detect MP rapidly and conveniently are still lacking. This study aimed to use the fluorescent dye Alexa Fluor® 647 as the detection marker to develop a lateral flow immunochromatographic assay (LFIA) for detection of MP-specific IgM in serum specimen. Monoclonal mouse antibody against human IgM (μ-chain specific) and goat anti-rabbit IgG were labeled with Alexa Fluor® 647 (anti-IgM-AF647 and anti-IgG-AF647). A mixture of natural MP antigen and recombinant P1 antigen was coated as the test line (T line) and rabbit IgG was coated as the control line (C line) on a nitrocellulose (NC) membrane. The MP antigens captured IgM-anti-IgM-AF647 complex on the T line. Rabbit IgG captured anti-IgG-AF647 on the C line. The fluorescence intensity on the T line and C line was measured. Sartorius CN140 NC membrane showed higher sensitivity than CN95. The optimal reaction time for the LFIA was 10min. The area under the receiver operating characteristic curve based on 34 MP positive and 166 MP negative serum samples was 0.986 (pLFIA strips did not react with serum from patients infected with non-MP pathogens including influenza viruses and bacteria causing respiratory tract infection. The intra-assay and inter-assay coefficients of variation were between 3.28% and 10.14%. The shelf life was calculated to be 2years at room temperature. The LFIA strips and the commercial EUROIMMUN kit showed consistent results on 372 serum specimens. The overall consistency rate was 96.37% with a Kappa value of 0.842 (pLFIA in the current study may be a sensitive and specific approach to detect early MP infection rapidly and conveniently. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Rapid detection of anti-Vaccinia virus neutralizing antibodies

    Directory of Open Access Journals (Sweden)

    Lichtfuss Gregor F

    2011-03-01

    Full Text Available Abstract Increasing infections with Monkeypox and Cowpox viruses pose a continuous and growing threat to human health. The standard method for detecting poxvirus neutralizing antibodies is the plaque-reduction neutralization test that is specific but also time-consuming and laborious. Therefore, a rapid and reliable method was developed to determine neutralizing antibody titers within twelve hours. The new assay measures viral mRNA transcription as a marker for actively replicating virus after incomplete neutralization using real-time PCR.

  4. Growing random networks with fitness

    OpenAIRE

    Ergun, G.; Rodgers, GJ

    2001-01-01

    Three models of growing random networks with fitness dependent growth rates are analysed using the rate equations for the distribution of their connectivities. In the first model (A), a network is built by connecting incoming nodes to nodes of connectivity $k$ and random additive fitness $\\eta$, with rate $(k-1)+ \\eta $. For $\\eta >0$ we find the connectivity distribution is power law with exponent $\\gamma=+2$. In the second model (B), the network is built by connecting nodes to nodes of conn...

  5. Neural-like growing networks

    Science.gov (United States)

    Yashchenko, Vitaliy A.

    2000-03-01

    On the basis of the analysis of scientific ideas reflecting the law in the structure and functioning the biological structures of a brain, and analysis and synthesis of knowledge, developed by various directions in Computer Science, also there were developed the bases of the theory of a new class neural-like growing networks, not having the analogue in world practice. In a base of neural-like growing networks the synthesis of knowledge developed by classical theories - semantic and neural of networks is. The first of them enable to form sense, as objects and connections between them in accordance with construction of the network. With thus each sense gets a separate a component of a network as top, connected to other tops. In common it quite corresponds to structure reflected in a brain, where each obvious concept is presented by certain structure and has designating symbol. Secondly, this network gets increased semantic clearness at the expense owing to formation not only connections between neural by elements, but also themselves of elements as such, i.e. here has a place not simply construction of a network by accommodation sense structures in environment neural of elements, and purely creation of most this environment, as of an equivalent of environment of memory. Thus neural-like growing networks are represented by the convenient apparatus for modeling of mechanisms of teleological thinking, as a fulfillment of certain psychophysiological of functions.

  6. Performance of the SD BIOLINE® HAT rapid test in various diagnostic algorithms for gambiense human African trypanosomiasis in the Democratic Republic of the Congo.

    Directory of Open Access Journals (Sweden)

    Crispin Lumbala

    Full Text Available We carried out a study to compare the performance, in terms of sensitivity and specificity, of the new SD BIOLINE® HAT rapid diagnostic test (RDT with the card agglutination test for trypanosomiasis (CATT for diagnosis of human African trypanosomiasis (HAT in the Democratic Republic of the Congo (DRC. Participants were enrolled actively by four mobile teams, and passively at four health facilities in three provinces. Consenting participants were tested concurrently with the RDT and CATT on whole blood. Those found positive by either test were tested with CATT on serial dilutions of plasma, and with a parasitological composite reference standard (CRS. Cases were only the individuals found positive by the CRS, while controls were negative by both CATT and RDT, as well as those that were positive by CATT or RDT, but were negative by the CRS, and had no history of HAT. Over five months, 131 cases and 13,527 controls were enrolled. The sensitivity of the RDT was 92.0% (95% confidence interval (CI = 86.1-95.5, which was significantly higher than CATT (sensitivity 69.1%; 95% CI = 60.7-76.4. The sensitivity of CATT on plasma at a dilution of 1:8 was 59.0% (95% CI = 50.2-67.2. The specificity of the RDT was 97.1% (95% CIs = 96.8-97.4 while that of CATT was 98.0% (95% CIs = 97.8, 98.2 and specificities of algorithms involving CATT at 1:8 dilution were 99.6% (95% CI = 99.5-99.7. Reproducibility of results was excellent. We concluded that an algorithm in which the SD BIOLINE® HAT RDT is used for screening is optimal for case detection in both passive and active screening settings. However, the lower specificity of the RDT compared to that of CATT would result in a larger number of false positive individuals undergoing confirmatory testing.

  7. Development and validation of a rapid and sensitive UPLC-MS/MS method for determination of uracil and dihydrouracil in human plasma.

    Science.gov (United States)

    Jacobs, Bart A W; Rosing, Hilde; de Vries, Niels; Meulendijks, Didier; Henricks, Linda M; Schellens, Jan H M; Beijnen, Jos H

    2016-07-15

    Quantification of the endogenous dihydropyrimidine dehydrogenase (DPD) substrate uracil (U) and the reaction product dihydrouracil (UH2) in plasma might be suitable for identification of patients at risk of fluoropyrimidine-induced toxicity as a result of DPD deficiency. In this paper, we describe the development and validation of a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay for quantification of U and UH2 in human plasma. Analytes were extracted by protein precipitation, chromatographically separated on an Acquity UPLC(®) HSS T3 column with gradient elution and analyzed with a tandem mass spectrometer equipped with an electrospray ionization source. U was quantified in the negative ion mode and UH2 in the positive ion mode. Stable isotopes for U and UH2 were used as internal standards. Total chromatographic run time was 5min. Validated concentration ranges for U and UH2 were from 1 to 100ng/mL and 10 to 1000ng/mL, respectively. Inter-assay bias and inter-assay precision for U were within ±2.8% and ≤12.4%. For UH2, inter-assay bias and inter-assay precision were within ±2.9% and ≤7.2%. Adequate stability of U and UH2 in dry extract, final extract, stock solution and plasma was demonstrated. Stability of U and UH2 in whole blood was only satisfactory when stored up to 4hours at 2-8°C, but not at ambient temperatures. An accurate, precise and sensitive UPLC-MS/MS assay for quantification of U and UH2 in plasma was developed. This assay is now applied to support clinical studies with fluoropyrimidine drugs. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. A rapid and sensitive HPLC method for the analysis of metronidazole in human plasma: application to single dose pharmacokinetic and bioequivalence studies

    Directory of Open Access Journals (Sweden)

    Jaber Emami

    2006-03-01

    Full Text Available A sensitive, accurate and rapid reverse phase HPLC method was developed to quantitate plasma levels of metronidazole in order to conduct a comparative bioavailability studies. The drug and internal standard were added to plasma samples, vortexed and then zinc sulfate solution was added in order to precipitate the plasma proteins. Samples were centrifuged at 3000 rpm for 10 min. The supernatant layer was separated and analyzed on a phenyl (300 × 4.6mm column, with 5% acetonitrile in 0.1 M KH2PO4 buffer (pH = 4.5 at 324 nm. The standard curve covering 0.15 – 30 μg/ml concentration range, was linear (r2 = 0.9999, relative errors were within 2.48 to 9.15 % and the CV% ranged from 2.999 to 10.796. The method is suitable for bioavailability, pharmacokinetic, and bioequivalent studies in human. The in-vivo study was carried out in 12 healthy volunteers according to a single dose, two-sequence, cross over randomized design. The bioavailability was compared using the total area under the plasma level versus time curve (AUC0-48, AUC0-, peak plasma concentration (Cmax and time to Cmax (Tmax. No statistically significant difference was found between the AUC0- , Cmax and Tmax values of the test and reference, Flagyl® (p > 0.05. The 90% CI for the ratio of the AUC0- (0.94-1.07 and Cmax (0.88-1.03 and the logarithmically transformed AUC0- (0.99-1.01 and Cmax (0.94-1.01 values of the generic product over those of Flagyl® was calculated to be within the acceptable limit of 0.80-1.20 and 0.80-1.25, respectively. It was, therefore, concluded that the generic metronidazole was bioequivalent with the innovator formulation.

  9. A rapid immunization strategy with a live attenuated tetravalent dengue vaccine elicits protective neutralizing antibody responses in non-human primates

    Directory of Open Access Journals (Sweden)

    Yuping eAmbuel

    2014-06-01

    Full Text Available Dengue viruses (DENVs cause approximately 390 million cases of DENV infections annually and over 3 billion people worldwide are at risk of infection. No dengue vaccine is currently available nor is there an antiviral therapy for DENV infections. We have developed a tetravalent live-attenuated DENV vaccine (TDV that consists of a molecularly characterized attenuated DENV-2 strain (TDV-2 and three chimeric viruses containing the pre-membrane and envelope genes of DENV-1, -3 and -4 expressed in the context of the TDV-2 genome. To impact dengue vaccine delivery in endemic areas and immunize travelers, a simple and rapid immunization strategy (RIS is preferred. We investigated RIS consisting of two full vaccine doses being administered subcutaneously or intradermally on the initial vaccination visit (day 0 at two different anatomical locations with a needle-free disposable syringe jet injection (DSJI delivery devices (PharmaJet in non-human primates (NHP. This vaccination strategy resulted in efficient priming and induction of neutralizing antibody responses to all four DENV serotypes comparable to those elicited by the traditional prime and boost (two months later vaccination schedule. In addition, the vaccine induced CD4+ and CD8+ T cells producing IFN-γ, IL-2, and TNF-α, and targeting the DENV-2 NS1, NS3 and NS5 proteins. Moreover, vaccine-specific T cells were cross-reactive with the non-structural NS3 and NS5 proteins of DENV-4. When animals were challenged with DENV-2 they were protected with no detectable viremia, and exhibited sterilizing immunity (no increase of neutralizing titers post- challenge. RIS could decrease vaccination visits and provide quick immune response to all four DENV serotypes. This strategy could increase vaccination compliance and would be especially advantageous for travelers into endemic areas.

  10. Rapid simultaneous determination of paracetamol, amantadine hydrochloride, caffeine and chlorpheniramine maleate in human plasma by liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Feng, Shudan; Tian, Yuan; Zhang, Zunjian; Zhang, Jingjing; Huang, Meihua; Chen, Yun

    2009-01-01

    A rapid, simple and sensitive high performance liquid chromatography with positive ion electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was first developed and validated to simultaneously determine paracetamol (PAR, CAS 103-90-2), amantadine hydrochloride (ATH, CAS 665-66-7), caffeine (CAF, CAS 58-08-2) and chlorpheniramine maleate (CPM, CAS 113-92-8) in human plasma using tramadol hydrochloride (TMH, CAS 22204-88-2) as internal standard (IS). Following methanol-induced protein precipitation, the analytes were separated using a mobile phase comprised of methanol:water (0.5% formic acid) = 20:80 (v/v) on a commercially available column (150 mm x 2.1 mm I.D., 5 microm) and analyzed by electrospray ionization tandem mass spectrometry in the selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 152.3-->110.2 for PAR, 152.3-->135.3 for ATH, 195.1-->138.3 for CAF, 275.2-->230.3 for CPM and 264.2-->58.2 for TMH. The standard curves were linear (r2 > 0.99) over the concentration range of 0.2-20 microg/mL for PAR, 20-2000 ng/mL for ATH and CAF, 0.1-10 ng/mL for CPM and had good accuracy and precision, respectively. The within- and between-batch precisions were less than 15% in terms of relative standard deviation (RSD). The lower limit of quantitation (LLOQ) were 0.2 microg/mL, 20 ng/mL, 20 ng/mL and 0.1 ng/mL for PAR, ATH, CAF and CPM, respectively. The described method has been successfully applied to study the pharmacokinetics of paracetamol-amantadine hydrochloride tablets in Chinese healthy male volunteers with great precision and sensitivity.

  11. Optimization and validation of a fast amplification protocol for AmpFlSTR® Profiler Plus® for rapid forensic human identification.

    Science.gov (United States)

    Laurin, Nancy; Frégeau, Chantal

    2012-01-01

    The goal of this work was to optimize and validate a fast amplification protocol for the multiplex amplification of the STR loci included in AmpFlSTR(®) Profiler Plus(®) to expedite human DNA identification. By modifying the cycling conditions and by combining the use of a DNA polymerase optimized for high speed PCR (SpeedSTAR™ HS) and a more efficient thermal cycler instrument (Bio-RAD C1000™), we were able to reduce the amplification process from 4h to 26 min. No modification to the commercial AmpFlSTR(®) Profiler Plus(®) primer mix was required. When compared to the current Royal Canadian Mounted Police (RCMP) amplification protocol, no differences with regards to specificity, sensitivity, heterozygote peak height ratios and overall profile balance were noted. Moreover, complete concordance was obtained with profiles previously generated with the standard amplification protocol and minor alleles in mixture samples were reliably typed. An increase in n-4 stutter ratios (2.2% on average for all loci) was observed for profiles amplified with the fast protocol compared to the current procedure. Our results document the robustness of this rapid amplification protocol for STR profiling using the AmpFlSTR(®) Profiler Plus(®) primer set and demonstrate that comparable data can be obtained in substantially less time. This new approach could provide an alternative option to current multiplex STR typing amplification protocols in order to increase throughput or expedite time-sensitive cases. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  12. [Transcriptional regulation effect of THSG and anthraquinones in tubers of Polygonum multiflorum based on human progesterone X receptor (PXR) mediated CYP3A4 rapid screening system].

    Science.gov (United States)

    Zhang, Zhao-Yan; Yang, Liang; Huang, Xiao-Yan; Wang, Mei-Xi; Ma, Zeng-Chun; Tang, Xiang-Lin; Wang, Yu-Guang; Gao, Yue

    2017-12-01

    The rapid screening technology was used to investigate the transcriptional regulation effect of main chemical constituents in tubers of Polygonum multiflorum, including 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside(THSG) and anthraquinones (such as rhein, chrysophanol, aloe-emodin, emodin) on CYP3A4 drug inducers induced by human pregnancy X receptor (PXR).The effect of chemical composition on the cell activity was detected by MTS cell viability assay. IC₅₀ was calculated. The expression vector and the reporter vector were co-transfected into HepG2 cells, with 10 μmol•L⁻¹ rifampicin (RIF) as a positive control, and 10 μmol•L⁻¹ ketoconazole (TKZ) as a negative control. After treated with different concentrations of anthraquinones (2.5, 5, 10 μmol•L⁻¹) for 24 h, the cells were tested for dual luciferase activity. The results show that the inhibitory effect of THSG, chrysophanol, emodin, rhein and aloe-emodin on CYP3A4 was inhibited by co-transfection of pcDNA3.1 and pGL4.17-CYP3A4. The expressions of pcDNA3.14-PXR and pGL4.17-CYP3A4 were induced by the four compounds. Besides, emodin had a direct inducing effect. In conclusion, the four anthraquinone compounds have an inducing effect on CYP3A4 by PXR, but emodin can directly induce CYP3A4. THSG can inhibit CYP3A4, but plasmid can induce CYP3A4 after intervened with PXR.These results suggest that we should pay attention to the liver function and avoid liver damage in the combined administration of drugs. Copyright© by the Chinese Pharmaceutical Association.

  13. [Myopia, a growing health problem

    NARCIS (Netherlands)

    Tideman, J.W.; Polling, J.R.; Schans, A. van der; Verhoeven, V.J.; Klaver, C.C.W.

    2016-01-01

    - Myopia is the eye disorder with the most rapid increase in prevalence worldwide. It develops in childhood, with a peak incidence between the ages of 13 to 15 years. - Especially high myopia, i.e. a refractive error of -6 diopters or more, increases the risk of permanent visual impairment during

  14. ISHAM-ITS reference DNA barcoding database - the quality controlled standard tool for routine identification of human and animal pathogenic fungi

    NARCIS (Netherlands)

    Robert, Vincent

    2015-01-01

    Human and animal fungal pathogens are a growing threat worldwide leading to emerging infections and creating new risks for established ones. There is a growing need for a rapid and accurate identification of pathogens to enable early diagnosis and targeted antifungal therapy. Morphological and

  15. Rapid transient expression of human granulocyte-macrophage colony-stimulating factor in two industrial cultivars of tobacco (Nicotiana tabacum L.) by agroinfiltration

    OpenAIRE

    Vojta, Lea; Ljuma-Skupnjak, Lana; Budimir, Ankica; Vukičević, Slobodan; Fulgosi, Hrvoje

    2015-01-01

    We report the production of hGM-CSF cytokine in leaves of industrial tobacco cultivars DH-17 and DH-27 by using Agrobacterium-mediated transient expression. We prove the concept that very high biomass industrial tobacco plants are suitable platforms for rapid, low cost production of foreign proteins. Successful transient expression of the GM-CSF was achieved in less than three months, opening the possibility for future applications of this approach in rapid response production of various prot...

  16. Human Neutrophil Lipocalin in Activated Whole Blood Is a Specific and Rapid Diagnostic Biomarker of Bacterial Infections in the Respiratory Tract.

    Science.gov (United States)

    Venge, Per; Eriksson, Ann-Katrin; Douhan-Håkansson, Lena; Pauksen, Karlis

    2017-07-01

    The distinction between bacterial and viral causes of infections of the respiratory tract is a major but important clinical challenge. We investigated the diagnostic performance of human neutrophil lipocalin (HNL) in respiratory tract infections compared to those of C-reactive protein (CRP) and procalcitonin (PCT). Patients were recruited from the emergency department and from a primary care unit (n = 162). The clinical diagnosis with regard to bacterial or viral cause of infection was complemented with objective microbiological/serological testing. HNL was measured in whole blood after preactivation with the neutrophil activator formyl-methionine-leucine-phenylalanine (fMLP) (B-HNL), and CRP and PCT were measured in plasma. Head-to-head comparisons of the three biomarkers showed that B-HNL was a superior diagnostic means to distinguish between causes of infections, with areas under the concentration-time curve (AUCs) of receiver operating characteristic (ROC) analysis for HNL of 0.91 (95% confidence interval [CI], 0.83 to 0.96) and 0.92 (95% CI, 0.82 to 0.97) for all respiratory infections and for upper respiratory infections, respectively, compared to 0.72 (95% CI, 0.63 to 0.80) and 0.68 (95% CI, 0.56 to 0.79) for CRP, respectively (P = 0.001). In relation to major clinical symptoms of respiratory tract infections (cough, sore throat, stuffy nose, and signs of sinusitis), AUCs varied between 0.88 and 0.93 in those patients with likely etiology (i.e., etiology is likely determined) of infection, compared to 0.63 and 0.71 for CRP, respectively, and nonsignificant AUCs for PCT. The diagnostic performance of B-HNL is superior to that of plasma CRP (P-CRP) and plasma PCT (P-PCT) in respiratory tract infections, and the activity specifically reflects bacterial challenge in the body. The rapid and accurate analysis of HNL by point-of-care technologies should be a major advancement in the diagnosis and management of respiratory infections with respect to antibiotic

  17. Transport of Nutrients Determines Growth in Tissue Culture; Why apple shoots grow rapidly and tulip shoots grow slowly

    NARCIS (Netherlands)

    Klerk, de G.J.M.

    2015-01-01

    Tulip growth in vitro is seriously impaired by inferior transport in the shoots. As a result, tulip cannot be micropropagated commercially using conventional means. In contrast, apple shoots show high transport and are easily micropropagated.

  18. Biofilm-growing intestinal anaerobic bacteria.

    Science.gov (United States)

    Donelli, Gianfranco; Vuotto, Claudia; Cardines, Rita; Mastrantonio, Paola

    2012-07-01

    Sessile growth of anaerobic bacteria from the human intestinal tract has been poorly investigated, so far. We recently reported data on the close association existing between biliary stent clogging and polymicrobial biofilm development in its lumen. By exploiting the explanted stents as a rich source of anaerobic bacterial strains belonging to the genera Bacteroides, Clostridium, Fusobacterium, Finegoldia, Prevotella, and Veillonella, the present study focused on their ability to adhere, to grow in sessile mode and to form in vitro mono- or dual-species biofilms. Experiments on dual-species biofilm formation were planned on the basis of the anaerobic strains isolated from each clogged biliary stent, by selecting those in which a couple of anaerobic strains belonging to different species contributed to the polymicrobial biofilm development. Then, strains were investigated by field emission scanning electron microscopy and confocal laser scanning microscopy to reveal if they are able to grow as mono- and/or dual-species biofilms. As far as we know, this is the first report on the ability to adhere and form mono/dual-species biofilms exhibited by strains belonging to the species Bacteroides oralis, Clostridium difficile, Clostridium baratii, Clostridium fallax, Clostridium bifermentans, Finegoldia magna, and Fusobacterium necrophorum. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  19. Growing the Blockchain information infrastructure

    DEFF Research Database (Denmark)

    Jabbar, Karim; Bjørn, Pernille

    2017-01-01

    In this paper, we present ethnographic data that unpacks the everyday work of some of the many infrastructuring agents who contribute to creating, sustaining and growing the Blockchain information infrastructure. We argue that this infrastructuring work takes the form of entrepreneurial actions......, which are self-initiated and primarily directed at sustaining or increasing the initiator’s stake in the emerging information infrastructure. These entrepreneurial actions wrestle against the affordances of the installed base of the Blockchain infrastructure, and take the shape of engaging...... or circumventing activities. These activities purposefully aim at either influencing or working around the enablers and constraints afforded by the Blockchain information infrastructure, as its installed base is gaining inertia. This study contributes to our understanding of the purpose of infrastructuring, seen...

  20. Growing Vertical in the Flatland.

    Science.gov (United States)

    Robinson, Joshua A

    2016-01-26

    The world of two-dimensional (2D) heterostructures continues to expand at a rate much greater than anyone could have predicted 10 years ago, but if we are to make the leap from science to technology, many materials challenges must still be overcome. Recent advances, such as those by Liu et al. in this issue of ACS Nano, demonstrate that it is possible to grow rotationally commensurate 2D heterostructures, which could pave the way toward single crystal van der Waals solids. In this Perspective, I provide some insight into a few of the challenges associated with growth of heterostructures, and discuss some of the recent works that help us better understand synthetic realization of 2D heterostructures.

  1. Viking Disruptions or Growing Integration?

    DEFF Research Database (Denmark)

    Sindbæk, Søren Michael

    2012-01-01

    Long-distance communication has emerged as a particular focus for archaeological exploration using network theory, analysis, and modelling. Initial attempts to adapt methods from social network analysis to archaeological data have, however, struggled to produce decisive results. This paper...... demonstrates how formal network analysis can be combined with a contextual reading of evidence relating to a long-distance communication network in the past. A study of the combined distributions of ten vessel types in 152 settlement sites from the 10th century suggests the outline of the core structure...... of the network. The model implies that 10th century long-distance exchange in the North Sea region featured long-distance links equal to those of the Carolingian emporia trade, and represented a growth in terms of new axes of integration, above all the growing links between the Scandinavian Peninsula...

  2. Growing bubbles rising in line

    Directory of Open Access Journals (Sweden)

    John F. Harper

    2001-01-01

    Full Text Available Over many years the author and others have given theories for bubbles rising in line in a liquid. Theory has usually suggested that the bubbles will tend towards a stable distance apart, but experiments have often showed them pairing off and sometimes coalescing. However, existing theory seems not to deal adequately with the case of bubbles growing as they rise, which they do if the liquid is boiling, or is a supersaturated solution of a gas, or simply because the pressure decreases with height. That omission is now addressed, for spherical bubbles rising at high Reynolds numbers. As the flow is then nearly irrotational, Lagrange's equations can be used with Rayleigh's dissipation function. The theory also works for bubbles shrinking as they rise because they dissolve.

  3. Morphogenesis of Growing Soft Tissues

    Science.gov (United States)

    Dervaux, Julien; Ben Amar, Martine

    2008-08-01

    Recently, much attention has been given to a noteworthy property of some soft tissues: their ability to grow. Many attempts have been made to model this behavior in biology, chemistry, and physics. Using the theory of finite elasticity, Rodriguez has postulated a multiplicative decomposition of the geometric deformation gradient into a growth-induced part and an elastic one needed to ensure compatibility of the body. In order to fully explore the consequences of this hypothesis, the equations describing thin elastic objects under finite growth are derived. Under appropriate scaling assumptions for the growth rates, the proposed model is of the Föppl von Kármán type. As an illustration, the circumferential growth of a free hyperelastic disk is studied.

  4. Impact of growing income inequality on sustainable development in China: a provincial-level analysis

    NARCIS (Netherlands)

    Heerink, N.B.M.; Ma, J.

    2006-01-01

    A growing body of literature has documented the rapidly increasing income disparities that accompanied China's economic growth in the 1980s and 1990s, and the driving factors behind this. Growing income inequality in its turn may have important implications for the accumulation of physical capital,

  5. Food Security Hotspots in India under Changing Climate and Growing Populatio

    Science.gov (United States)

    Singh, N.; Cherkauer, K. A.; Ashfaq, M.; Rastogi, D.

    2015-12-01

    Global climate change, unprecedented population growth, and rapid urbanization have elevated the possibility of food and water shortages in many regions across the world. The likelihood of such shortages in the future is further exacerbated by the fact that increased greenhouse forcing and rapid growth in human population will continue for at least the next several decades. These socio-environmental changes will likely put some regions under enormous economic and environmental stress by forcing them to adapt to new conditions. India with its rapidly growing population and high rates of urbanization and industrialization is one such region whose agricultural resources will be particularly vulnerable to the impact of these changes. This study collectively reviews and analyses the possible impacts of climate change, population growth and resulting land use change on the availability of food and water in the coming decades for India. By analyzing and fusing a wide variety of existing data and research on the impact of land use change, population, and climate change, on water and food resources this study develops an understanding of the broader implications of each of the changes on food security in the region. In addition, the study focuses on the need to assess and quantify the combination of such impacts at a regional level and identify food security hotspots spatially across India that will help to narrow down regions in the country which will be severely affected and need priority adaptation and mitigation measures.

  6. Rapid transient expression of human granulocyte-macrophage colony-stimulating factor in two industrial cultivars of tobacco (Nicotiana tabacum L. by agroinfiltration

    Directory of Open Access Journals (Sweden)

    Lea Vojta

    2015-09-01

    Full Text Available We report the production of hGM-CSF cytokine in leaves of industrial tobacco cultivars DH-17 and DH-27 by using Agrobacterium-mediated transient expression. We prove the concept that very high biomass industrial tobacco plants are suitable platforms for rapid, low cost production of foreign proteins. Successful transient expression of the GM-CSF was achieved in less than three months, opening the possibility for future applications of this approach in rapid response production of various proteins of non-plant origin in industrial tobacco.

  7. [Myopia, a growing health problem].

    Science.gov (United States)

    Tideman, J W L; Polling, J R; van der Schans, A; Verhoeven, V J M; Klaver, C C W

    2016-01-01

    - Myopia is the eye disorder with the most rapid increase in prevalence worldwide. It develops in childhood, with a peak incidence between the ages of 13 to 15 years. - Especially high myopia, i.e. a refractive error of -6 diopters or more, increases the risk of permanent visual impairment during adulthood due to structural abnormalities of the retina and optic nerve.- The cause of myopia is complex. Lifestyle factors in childhood, such as limited time spent outdoors and close work - such as reading and smartphone usage - are risk factors. Furthermore, genetic studies have revealed more than 100 factors associated with the development of myopia. - Pharmacological and optical interventions to inhibit myopia progression are increasingly applied. The use of atropine eye drops in children and has shown to be an effective treatment.

  8. Infection Dynamics on Growing Networks

    Science.gov (United States)

    Lai, Ying-Cheng; Liu, Zonghua; Ye, Nong

    We consider the entire spectrum of architectures for large, growing, and complex networks, ranging from being heterogeneous (scale-free) to homogeneous (random or small-world), and investigate the infection dynamics by using a realistic three-state epidemiological model. In this framework, a node can be in one of the three states: susceptible (S), infected (I), or refractory (R), and the populations in the three groups are approximately described by a set of nonlinear differential equations. Our heuristic analysis predicts that, (1) regardless of the network architecture, there exists a substantial fraction of nodes that can never be infected, and (2) heterogeneous networks are relatively more robust against spread of infection as compared with homogeneous networks. These are confirmed numerically. We have also considered the problem of deliberate immunization for preventing wide spread of infection, with the result that targeted immunization can be quite effective for heterogeneous networks. We believe these results are important for a host of problems in many areas of natural science and engineering, and in social sciences as well.

  9. Growing plants on atoll soils

    Energy Technology Data Exchange (ETDEWEB)

    Stone, E L; Migvar, L; Robison, W L

    2000-02-16

    Many years ago people living on atolls depended entirely on foods gathered from the sea and reefs and grown on land. Only a few plants, such as coconut (ni), Pandanus (bob), and arrowroot (mok-mok), could be grown on the lower rainfall atolls, although adequate groundwater conditions also allowed taro (iaraj, kotak, wot) to be cultivated. On higher rainfall atolls, breadfruit (ma) was a major food source, and banana (binana, kepran), lime (laim), and taros (iaraj, kotak, wot) could be grown. The early atoll populations were experts in growing plants that were vital to sustaining their nutrition requirements and to providing materials for thatch, basketry, cordage, canoe construction, flowers, and medicine. They knew which varieties of food plants grew well or poorly on their atolls, how to propagate them, and where on their atoll they grew best. They knew the uses of most native plants and what the various woods were well suited for. Many varieties of Pandanus (bob) and breadfruit (ma) grew well with high rainfall, but only a few produced well on drier atolls. Such information had been passed down through the generations although some of it has been lost in the last century. Today there are new plants and new varieties of existing plants that can be grown on atolls. There are also new materials and information on how to grow both the old and new plants more effectively. However, there are also introduced weeds and pests to control. Today, there is also an acute need to grow more of the useful plants adapted to atolls. Increasing numbers of people living on an atoll without an equal increase in income or food production stretches the available food supplies. Much has been written about the poor conditions for plant growth on atolls. As compared with many places in the world where crops are grown, however, atolls can provide some highly favorable conditions. For instance, the driving force for plant growth is sunlight, and on atolls light is abundant throughout the

  10. Growing and evolving soft robots.

    Science.gov (United States)

    Rieffel, John; Knox, Davis; Smith, Schuyler; Trimmer, Barry

    2014-01-01

    Completely soft and flexible robots offer to revolutionize fields ranging from search and rescue to endoscopic surgery. One of the outstanding challenges in this burgeoning field is the chicken-and-egg problem of body-brain design: Development of locomotion requires the preexistence of a locomotion-capable body, and development of a location-capable body requires the preexistence of a locomotive gait. This problem is compounded by the high degree of coupling between the material properties of a soft body (such as stiffness or damping coefficients) and the effectiveness of a gait. This article synthesizes four years of research into soft robotics, in particular describing three approaches to the co-discovery of soft robot morphology and control. In the first, muscle placement and firing patterns are coevolved for a fixed body shape with fixed material properties. In the second, the material properties of a simulated soft body coevolve alongside locomotive gaits, with body shape and muscle placement fixed. In the third, a developmental encoding is used to scalably grow elaborate soft body shapes from a small seed structure. Considerations of the simulation time and the challenges of physically implementing soft robots in the real world are discussed.

  11. Pediatric Ovarian Growing Teratoma Syndrome

    Directory of Open Access Journals (Sweden)

    Rebecca M. Rentea

    2017-01-01

    Full Text Available Ovarian immature teratoma is a germ cell tumor that comprises less than 1% of ovarian cancers and is treated with surgical debulking and chemotherapy depending on stage. Growing teratoma syndrome (GTS is the phenomenon of the growth of mature teratoma elements with normal tumor markers during or following chemotherapy for treatment of a malignant germ cell tumor. These tumors are associated with significant morbidity and mortality due to invasive and compressive growth as well as potential for malignant transformation. Current treatment modality is surgical resection. We discuss a 12-year-old female who presented following resection of a pure ovarian immature teratoma (grade 3, FIGO stage IIIC. Following chemotherapy and resection of a pelvic/liver recurrence demonstrating mature teratoma, she underwent molecular genetics based chemotherapeutic treatment. No standardized management protocol has been established for the treatment of GTS. The effect of chemotherapeutic agents for decreasing the volume of and prevention of expansion is unknown. We review in detail the history, diagnostic algorithm, and previous reported pediatric cases as well as treatment options for pediatric patients with GTS.

  12. [Growing old differently: Transdisciplinary perspective].

    Science.gov (United States)

    Zimmermann, H-P

    2015-04-01

    Growing old differently: the phrase is intended to call something other to mind than merely the fact that images and forms of old age and aging have multiplied and diversified to an enormous extent. The suggestion put forward here is that otherness (as opposed to mere differences) should be positively reinforced. In other words, it is not just a matter of noting different forms of old age and aging but more than this, of seeking out opportunities for aging differently. In order to explore this, the article follows an older strand of theory, which has recently come to be frequently quoted in gerontology: the phenomenology of difference as reasoned analytically by Lévinas and Sartre and applied to gerontology by Améry and de Beauvoir. Here, opportunities for aging crucially depend on the way we look at it, how we observe and describe it and not least, how gerontology frames it. A distinction is made between two perspectives and their associated consequences for old age: alienation and alterity. Alienation means looking at old age above all as a disconcerting "other", as a perplexing, problematic deviation from the norm of vitality. Alterity, by contrast, refers to different options for living life in old age: options to be explored and opened up in contradistinction to cultural or academic alienation. Not least, the article appeals for diversity in scholarly approaches and for cross-disciplinary perspectives.

  13. Rapid and visual detection of human enterovirus coxsackievirus A16 by reverse transcription loop-mediated isothermal amplification combined with lateral flow device.

    Science.gov (United States)

    Yan, G; Jun, L; Kangchen, Z; Yiyue, G; Yang, Y; Xiaoyu, Z; Zhiyang, S; Lunbiao, C

    2015-12-01

    In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow device (LFD) technology to rapidly detect CVA16 was developed and evaluated. RT-LAMP assay was optimized to amplify VP1 gene of CVA16. Amplified products were analysed by LFD and capillary electrophoresis. The RT-LAMP-LFD assay showed 100% specificity in detecting CVA16, and showed analytical sensitivity of 0·55 TCID50 per reaction mixture. Comparison of the RT-LAMP-LFD assay with real-time RT-PCR developed previously in clinical specimens showed 93·3% agreement. The RT-LAMP-LFD assay is more sensitive in detecting CVA16 RNA. The RT-LAMP-LFD assay presented here might offer a rapid and simple alternative in clinical diagnosis of CVA16. Coxsackievirus A16 (CVA16) is one of the major causative agents of hand, foot and mouth disease (HFMD). Rapid and reliable detection and typing of it can limit the spread. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with lateral flow device (LFD) technology to rapidly detect CVA16. The high sensitivity and specificity and its ease of use make this assay ideal for use in resource-limited settings such as primary care facilities and clinical laboratories in developing countries. © 2015 The Society for Applied Microbiology.

  14. Rapid Population Growth and Human Carrying Capacity: Two Perspectives. World Bank Staff Working Papers No. 690 and Population and Development Series No. 15.

    Science.gov (United States)

    Mahar, Dennis J., Ed.; And Others

    Two perspectives on carrying capacity and population growth are examined. The first perspective, "Carrying Capacity and Rapid Population Growth: Definition, Cases, and Consequences" (Robert Muscat), explores the possible meanings of the idea of carrying capacity under developing country conditions, looks at historical and present-day cases of…

  15. Menopausal women's positive experience of growing older

    DEFF Research Database (Denmark)

    Hvas, Lotte

    2006-01-01

    This paper aims to describe menopausal women's positive experience of growing older and becoming middle-aged.......This paper aims to describe menopausal women's positive experience of growing older and becoming middle-aged....

  16. Bariatric amputee: A growing problem?

    Science.gov (United States)

    Kulkarni, Jai; Hannett, Dominic P; Purcell, Steven

    2015-06-01

    This study reviewed prevalence of patients with lower limb amputations with above normal weight profile, with body mass index over 25, in seven disablement services centres managing their amputee rehabilitation in the United Kingdom. To review two clinical standards of practice in amputee rehabilitation. Ambulant lower limb amputees should have their body weight recorded on an electronic information system, with identification of cohort with body weight >100 kg. Lower limb amputees to be provided with suitable weight-rated prosthesis. Observational study of clinical practice. Data were collected from the Clinical Information Management Systems. Inclusion criteria--subjects were ambulant prosthetic users with some prosthetic intervention in the last 5 years and had at least one lower limb amputation. In 96% of patients, the weight record profile was maintained. In addition, 86% were under 100 kg, which is the most common weight limit of prosthetic componentry. Of 15,204 amputation levels, there were 1830 transfemoral and transtibial sites in users with body weight over 100 kg. In 60 cases, the prosthetic limb build was rated to be below the user body weight. In 96% of our patients, body weight was documented, and in 97%, the prosthetic limb builds were within stated body weight limits, but this may not be the case in all the other disablement services centres in the United Kingdom. Also, the incidence of obesity in the United Kingdom is a growing problem, and the health issues associated with obesity are further compounded in the amputee population. Prosthetic componentry has distinct weight limits which must be considered during prescription. As people with amputation approach the limits of specific components, clinicians are faced with the challenge of continued provision in a safe and suitable manner. This article reviews the amputee population and the current national profile to consider trends in provision and the incidence of these challenges. © The

  17. Replication-defective recombinant Semliki Forest virus encoding GM-CSF as a vector system for rapid and facile generation of autologous human tumor cell vaccines

    NARCIS (Netherlands)

    Withoff, S; Glazenburg, KL; van Veen, ML; Kraak, MMJ; Hospers, GAP; Storkel, S; de Vries, EGE; Wischut, J; Daemen, T

    2001-01-01

    This paper describes the production of recombinant Semliki Forest virus encoding murine or human granulocyte-macrophage colony-stimulating factor (GM-CSF) and the capacity of these vectors to transduce murine and human tumor cells ex vivo. High-titer stocks (up to 3 x 10(9) particles/ml) of

  18. Cells growing in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

  19. Rapid simultaneous determination of eperisone, tolperisone, and tizanidine in human serum by using a MonoSpin® C18 extraction column and liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Miura, Naoya; Saito, Takeshi; Taira, Takayuki; Yamagiwa, Takeshi; Morita, Sein; Inokuchi, Sadaki

    2014-01-01

    A method was developed for rapid toxicological analysis of eperisone, tolperisone, and tizanidine in human serum using a MonoSpin® C18 extraction column and LC/MS/MS. The method was validated for LOD, linearity, precision, and extraction recovery. This method was rapid with an LOD of 0.5 ng/mL, linearity range 1-500.0 ng/mL (r2 = 0.999), and RSD value below 14.6%. Extraction recovery from the sample was greater than 98.6, 98.8, and 88.5% for eperisone, tolperisone, and tizanidine, respectively. Results showed that combination of the MonoSpin C18 extraction column and LC/MS/MS is a simple and rapid method for the analysis of these three analytes, and a method is described for simultaneous quantitative determination of the analytes in human serum by LC/MSIMS. This method was used to determine the serum levels of eperisone in a patient with eperisone poisoning, and could be successfully applied for screening analyses in clinical cases other than poisoning.

  20. Rapid detection of new and expanding human settlements in the Limpopo province of South Africa using a spatio-temporal change detection method

    CSIR Research Space (South Africa)

    Kleynhans, W

    2015-08-01

    Full Text Available Recent development has identified the benefits of using hyper-temporal satellite time series data for land cover change detection and classification in South Africa. In particular, the monitoring of human settlement expansion in the Limpopo province...

  1. Estradiol rapidly induces the translocation and activation of the intermediate conductance calcium activated potassium channel in human eccrine sweat gland cells.

    LENUS (Irish Health Repository)

    Muchekehu, Ruth W

    2009-02-01

    Steroid hormones target K+ channels as a means of regulating electrolyte and fluid transport. In this study, ion transporter targets of Estradiol (E2) were investigated in the human eccrine sweat gland cell line NCL-SG3.

  2. PD0332991 (palbociclib) for treatment of pediatric intracranial growing teratoma syndrome.

    Science.gov (United States)

    Schultz, Kris Ann P; Petronio, Joseph; Bendel, Anne; Patterson, Richard; Vaughn, David J

    2015-06-01

    Growing teratoma syndrome is characterized by growth of mature teratoma elements of a mixed germ cell tumor despite resolution of immature/malignant elements with administration of chemotherapy. Surgical resection is the only known cure for growing teratoma syndrome but in the brain, complete resection may be impossible. In these instances, mature teratoma, although histologically benign, may be fatal. In this report, we present the case of a child with a large, rapidly growing, unresectable pineal region growing teratoma. PD0332991 was administered with stabilization of the solid, enhancing components of the mass. Minimal adverse effects were noted. © 2014 Wiley Periodicals, Inc.

  3. Performance appraisal of rapid on-site malaria diagnosis (ICT malaria Pf/Pv test) in relation to human resources at village level in Myanmar.

    Science.gov (United States)

    Cho-Min-Naing; Gatton, M L

    2002-01-01

    Logistic, economic and technical factors limit rapid access to microscopic confirmation of symptomatic diagnosis of malaria in many rural areas in endemic countries such as Myanmar. A study was conducted to evaluate a rapid on-site immunochromatographic test (ICT Malaria Pf/Pv) for detection of Plasmodium falciparum and P. vivax in two villages in the Taikkyi region of Myanmar. The ICT Malaria tests were performed by a volunteer health worker (VHW) in Yae-Aye-San village and by a professionally trained midwife (MW) in Kankone village. A total of 1000 symptomatic patients participated in the study by providing blood samples for an ICT test and for microscopy. The ICT performance indices, relative to microscopy, were better for the trained MW compared with the less experienced VHW. For P. falciparum and/or P. vivax infections, the sensitivities were 82.7% for the VHW compared with 93.7% for the MW. For P. falciparum infections, the sensitivities were 82.2% for the VHW and 91.3% for the MW, while the corresponding values for P. vivax infections were 66.7 and 79%, respectively. Although the test kit appeared to perform better in more experienced hands, this study questions whether this difference is related to the use of the ICT Malaria Pf/Pv test kit, or related to other factors such as differences in the quality of blood slides prepared by the VHW and MW for microscopic examination. Overall, the results suggest that a rapid diagnostic assay such as the ICT Malaria Pf/Pv test kit can be used in rural settings by relatively inexperienced persons, such as VHWs, with a reasonable degree of sensitivity, thus providing on-site confirmation of symptomatic diagnosis of malaria.

  4. Can we grow new retina?

    Directory of Open Access Journals (Sweden)

    Geeta K Vemuganti

    2006-03-01

    Full Text Available Introduction Retinal degenerations and dystrophies, the major causes of genetically inherited blindness, are characterised by the death or degeneration of photoreceptors (rods and/or cones.1 Approaches to treating this disease include: a replacing the defective gene; b introducing a drug or agent that either slows down or stops the premature death of photoreceptors; c introducing electronic chips; or d replacing the damaged cells by cellular therapy. Gene therapy is aimed at counteracting the defective gene by substituting it with the normal gene in the target tissues. Though successful visual recovery has been reported with gene therapy in dog models,2-3 it remains a challenge to identify a safe and reliable way of introducing the corrective gene in humans, given that the genes need to act for the lifetime of the patient. Introduction of ‘a’ factors (such as growth factors into the eye, directly or through implants, is another novel approach to preventing or slowing premature cell death.4-5 The challenge lies in delivering the drug to the appropriate site in a safe and sustained manner. Electronic chips, similar to the ones used for audio aids, have shown exciting results in some studies, but the technology is still in its infancy.6-7

  5. A simple, rapid and validated high-performance liquid chromatography method suitable for clinical measurements of human mercaptalbumin and non-mercaptalbumin.

    Science.gov (United States)

    Yasukawa, Keiko; Shimosawa, Tatsuo; Okubo, Shigeo; Yatomi, Yutaka

    2018-01-01

    Background Human mercaptalbumin and human non-mercaptalbumin have been reported as markers for various pathological conditions, such as kidney and liver diseases. These markers play important roles in redox regulations throughout the body. Despite the recognition of these markers in various pathophysiologic conditions, the measurements of human mercaptalbumin and non-mercaptalbumin have not been popular because of the technical complexity and long measurement time of conventional methods. Methods Based on previous reports, we explored the optimal analytical conditions for a high-performance liquid chromatography method using an anion-exchange column packed with a hydrophilic polyvinyl alcohol gel. The method was then validated using performance tests as well as measurements of various patients' serum samples. Results We successfully established a reliable high-performance liquid chromatography method with an analytical time of only 12 min per test. The repeatability (within-day variability) and reproducibility (day-to-day variability) were 0.30% and 0.27% (CV), respectively. A very good correlation was obtained with the results of the conventional method. Conclusions A practical method for the clinical measurement of human mercaptalbumin and non-mercaptalbumin was established. This high-performance liquid chromatography method is expected to be a powerful tool enabling the expansion of clinical usefulness and ensuring the elucidation of the roles of albumin in redox reactions throughout the human body.

  6. Rapid Genotyping of the Human Renin (REN Gene by the LightCycler® Instrument: Identification of Unexpected Nucleotide Substitutions within the Selected Hybridization Probe Area

    Directory of Open Access Journals (Sweden)

    Line Wee

    2010-01-01

    Full Text Available Preeclampsia is a serious disorder affecting nearly 3% of all in the Western world. It is associated with hypertension and proteinuria, and several lines of evidence suggest that the renin-angiotensin system (RAS may be involved in the development of hypertension at different stages of a preeclamptic pregnancy. In this study, we developed rapid genotyping assays on the LightCycler® instrument to allow the detection of genetic variants in the renin gene (REN that may predispose to preeclampsia. The method is based on real-time PCR and allele-specific hybridization probes, followed by fluorescent melting curve analysis to expose a change in melting temperature (Tm. Ninety-two mother-father-child triads (n=276 from preeclamptic pregnancies were genotyped for three haplotype-tagging single nucleotide polymorphisms (htSNPs in REN. All three htSNPs (rs5705, rs1464816 and rs3795575 were successfully genotyped. Furthermore, two unexpected nucleotide substitutions (rs11571084 and rs61757041 were identified within the selected hybridization probe area of rs1464816 and rs3795575 due to aberrant melting peaks. In conclusion, genotyping on the LightCycler® instrument proved to be rapid and highly reproducible. The ability to uncover additional nucleotide substitutions is particularly important in that it allows the identification of potentially etiological variants that might otherwise be overlooked by other genotyping methods.

  7. In vivo and ex vivo methods of growing a liver bud through tissue connection.

    Science.gov (United States)

    Yanagi, Yusuke; Nakayama, Koichi; Taguchi, Tomoaki; Enosawa, Shin; Tamura, Tadashi; Yoshimaru, Koichiro; Matsuura, Toshiharu; Hayashida, Makoto; Kohashi, Kenichi; Oda, Yoshinao; Yamaza, Takayoshi; Kobayashi, Eiji

    2017-10-26

    Cell-based therapy has been proposed as an alternative to orthotopic liver transplantation. The novel transplantation of an in vitro-generated liver bud might have therapeutic potential. In vivo and ex vivo methods for growing a liver bud are essential for paving the way for the clinical translation of liver bud transplantation. We herein report a novel transplantation method for liver buds that are grown in vivo involving orthotopic transplantation on the transected parenchyma of the liver, which showed long engraftment and marked growth in comparison to heterotopic transplantation. Furthermore, this study demonstrates a method for rapidly fabricating scalable liver-like tissue by fusing hundreds of liver bud-like spheroids using a 3D bioprinter. Its system to fix the shape of the 3D tissue with the needle-array system enabled the fabrication of elaborate geometry and the immediate execution of culture circulation after 3D printing-thereby avoiding an ischemic environment ex vivo. The ex vivo-fabricated human liver-like tissue exhibited self-tissue organization ex vivo and engraftment on the liver of nude rats. These achievements conclusively show both in vivo and ex vivo methods for growing in vitro-generated liver buds. These methods provide a new approach for in vitro-generated liver organoids transplantation.

  8. Prevention and Care Programs Addressing the Growing Prevalence of Diabetes in China.

    Science.gov (United States)

    Yin, Junmei; Kong, Alice P S; Chan, Juliana C N

    2016-12-01

    According to a 2010 national survey, 11 % of adults in China have diabetes, affecting 109.6 million individuals. The high prevalence of diabetes has been attributed to the aging of the population, the rapid adoption of energy-dense foods, and a reduction in physical activity. Collectively, these secular changes have created an obesogenic environment that can unmask diabetes in subjects with a genetic predisposition. The growing prevalence of maternal obesity, gestational diabetes, childhood obesity, and early-onset disease can lead to premature morbidity and mortality. Rising to meet these public health challenges, researchers in China have conducted randomized studies to demonstrate the benefits of lifestyle modification in preventing diabetes (the Da Qing Study), as well as that of team-based integrated care, using multiple strategies including peer support and information technology, in order to reduce hospitalizations, cardiovascular-renal complications, and premature deaths. With growing evidence supporting the benefits of these diabetes prevention and management programs, the next challenge is to use policies and systems to scale up the implementation of these programs through raising awareness, building capacity, and providing resources to reduce the human and socioeconomic burden of diabetes.

  9. Growing pains in South America.

    Science.gov (United States)

    Joyce, S

    1997-08-01

    This article describes some negative effects from modernization and urban growth in South America, including disease, pesticides, occupational hazards, poor environmental controls of water and garbage, sanitation, and environmental degradation. South America is following the global trend toward urbanization and the problems that accompany it. Agricultural expansion led to an expanded market for pesticides that includes the deadly DDT, paraquat, and heptachlor. Brazil and Colombia are the largest consumers. Latin American officials justify use of DDT, which is banned in the US and many European countries, as an effective means of combating mosquitos that carry malaria. Exposure occurs during harvesting, transporting, forestry, livestock farming, and vector control activities. Methyl bromide, which is used post-harvest and as a soil fumigant, is dangerous enough to be banned in the US in 2001, and in developing countries in 2002. Exposure to toxic chemicals can severely inhibit enzyme action that is necessary for neurological functioning. A hot climate, which prevents protective clothing, lack of education on proper application, and absence of water to wash exposed skin, make pesticide protection very difficult. Over 40 million agricultural workers are at risk of pesticide poisoning. Habitat destruction has contributed to increased mosquito infestations. Children in the workplace are at even greater risk of noise pollution and chemical poisoning. South America pollutes almost 11 times more fresh water per capita than Europe. About 70% of domestic garbage is collected, and about 30% is disposed of correctly. Only 10% of urban wastewater is treated before discharge into waterways. The loss of coastal wetlands reduces the ability of waterways to filter and absorb nutrients. Environmental health problems suggest an interlinkage between environmental sustainability, human health, and economic growth.

  10. Amplification and expression of a cellular oncogene (c-myc) in human gastric adenocarcinoma cells.

    Science.gov (United States)

    Shibuya, M; Yokota, J; Ueyama, Y

    1985-01-01

    Three of 16 human gastric adenocarcinoma samples, maintained as solid tumors in nude mice, were found to carry amplified c-myc genes. In two samples with a high degree of c-myc DNA amplification (15- to 30-fold), double minute chromosomes were observed in karyotype analysis. The level of c-myc RNA was markedly elevated in a rapidly growing and poorly differentiated tumor, whereas it was only slightly elevated in a slowly growing and more differentiated tumor. Images PMID:2579323

  11. A rapid, sensitive and validated method for the determination of ondansetron in human plasma by reversed-phase high-pressure liquid chromatography.

    Science.gov (United States)

    Chandrasekar, Durairaj; Ramakrishna, Sistla; Diwan, Prakash V

    2004-01-01

    A simple and sensitive method for the determination of ondansetron (CAS 116002-70-1) in human plasma was developed using high-pressure liquid chromatography (HPLC). The procedure involves extraction of human plasma with tertiary butyl methyl ether containing 2 mol/l sodium hydroxide, followed by reversed-phase HPLC using a LiChrospher 100 RP-18e 5 microm column and UV detection at 305 nm. The retention times of ondansetron and internal standard (propranolol hydrochloride, CAS 318-98-9) were 9.38 and 13.40 min, respectively. The calibration curves were linear over the range of 10 ng/ml (lower limit of quantitation, LOQ) and 380 ng/ml for ondansetron. The intra- and inter-assay coefficients of variation for all the criteria of validation were less than 15% over the linearity range. Ondansetron was stable upon storage in human plasma. The sensitivity and precision of the method were within the accepted limits (< 15 %) throughout the validation period. The present method is useful for determination of plasma concentrations of ondansetron during human pharmacokinetic studies.

  12. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens

    NARCIS (Netherlands)

    Schuurman, T.; Lankamp, P.; van Belkum, A.; Kooistra-Smid, M.; van Zwet, A.

    2007-01-01

    Giardia lamblia is one of the most common intestinal parasites worldwide, with microscopy being the diagnostic reference standard for use with human stools. However, microscopy is time-consuming, labour-intensive and lacks sensitivity when single stools are examined. In the present study,

  13. Rapid, sensitive, type specific PCR detection of the E7 region of human papillomavirus type 16 and 18 from paraffin embedded sections of cervical carcinoma

    DEFF Research Database (Denmark)

    Lesnikova, Iana; Lidang, Marianne; Hamilton-Dutoit, Steven

    2010-01-01

    ABSTRACT: Human papillomavirus (HPV) infection, and in particularly infection with HPVs 16 and 18, is a central carcinogenic factor in the uterine cervix. We established and optimized a PCR assay for the detection and discrimination of HPV types 16 and 18 in archival formaldehyde fixed and paraffin...

  14. Rapid, sensitive, type specific PCR detection of the E7 region of human papillomavirus type 16 and 18 from paraffin embedded sections of cervical carcinoma

    DEFF Research Database (Denmark)

    Lesnikova, Iana; Lidang, Marianne; Hamilton-Dutoit, Stephen Jacques

    2010-01-01

    ABSTRACT: Human papillomavirus (HPV) infection, and in particularly infection with HPVs 16 and 18 is a central carcinogenic factor in the uterine cervix. We established and optimized a PCR assay for the detection and discrimination of HPV types 16 and 18 in archival formaldehyde fixed and paraffin...

  15. Micellar Enhanced Three-Dimensional Excitation-Emission Matrix Fluorescence for Rapid Determination of Antihypertensives in Human Plasma with Aid of Second-Order Calibration Methods

    Directory of Open Access Journals (Sweden)

    Hai-Yan Fu

    2015-01-01

    Full Text Available A highly sensitive three-dimensional excitation-emission fluorescence method was proposed to determine antihypertensives including valsartan and amlodipine besylate in human plasma with the aid of second-order calibration methods based on parallel factor analysis (PARAFAC and alternating trilinear decomposition (ATLD algorithms. Antihypertensives with weak fluorescent can be transformed into a strong fluorescent property by changing microenvironment in samples using micellar enhanced surfactant. Both the adopted algorithms with second-order advantage can improve the resolution and directly attain antihypertensives concentration even in the presence of potential strong intrinsic fluorescence from human plasma. The satisfactory results can be achieved for valsartan and amlodipine besylate in complicated human plasma. Furthermore, some statistical parameters and figures of merit were evaluated to investigate the performance of the proposed method, and the accuracy and precision of the proposed method were also validated by the elliptical joint confidence region (EJCR test and repeatability analysis of intraday and interday assay. The proposed method could not only light a new avenue to directly determine valsartan or amlodipine besylate in human plasma, but also hold great potential to be extended as a promising alternative for more practical applications in the determination of weak fluorescent drugs.

  16. Rapidly boosted Plasma IL-5 induced by treatment of human Schistosomiasis haematobium is dependent on antigen dose, IgE and eosinophils

    DEFF Research Database (Denmark)

    Wilson, Shona; Jones, Frances M.; Fofana, Hassan K. M.

    2013-01-01

    IgE specific to worm antigen (SWA) and pre-treatment eosinophil number, are associated with human immunity to re-infection with schistosomes after chemotherapeutic treatment. Treatment significantly elevates circulating IL-5 24-hr post-treatment of Schistosoma mansoni. Here we investigate if praz...

  17. Rapid opiate detoxification and antagonist induction under general anaesthesia or intravenous sedation is humane, sometimes essential and should always be an option. Three illustrative case reports involving diabetes and epilepsy and a review of the literature.

    Science.gov (United States)

    Brewer, Colin; de Jong, Catherine; Williams, Jonathan

    2014-01-01

    When abstinence is an appropriate goal, controlled studies and systematic reviews confirm that rapid, antagonist-precipitated opiate withdrawal procedures are the most effective and cost effective methods of initiating abstinence, and naltrexone (NTX) maintenance. While 'rapid' withdrawal, better conceptualised as Rapid Antagonist Induction (RAI), can often be humanely achieved with modest sedation levels, we present three case histories to support our argument that for some patients, general anaesthesia (GA), or techniques of intravenous sedation (IVS) that approach GA, are essential for safety and success. This includes patients with intercurrent disease (e.g. epilepsy or insulin-dependent diabetes) but also those with severe withdrawal phobia after previous distressing experiences. We discuss the history of the procedure. The dangers of RAI under GA or IVS in experienced hands have been exaggerated and the appropriate expertise should be more easily available. Patients and clinicians readily accept risks of major surgery for the excessive intake of food that causes most obesity. Similar risk-acceptance exists in cosmetic surgery and obstetrics. The increasing use and effectiveness of long-acting implants or depot-injections of NTX for relapse-prevention have largely solved compliance problems that undermined the potential of oral NTX. Their ability to prevent opiate overdose in abstinent, non-tolerant patients also strengthens arguments both for offering RAI as a therapeutic option and for reducing psychological, professional and practical barriers to using it.

  18. An enzyme-linked immuno focus assay for rapid detection and enumeration, and a newborn mouse model for human non-polio enteroviruses associated with acute diarrhea.

    Science.gov (United States)

    Rao, C Durga; Reddy, Harikrishna; Naidu, Jagadish R; Raghavendra, A; Radhika, N S; Karande, Anjali

    2015-11-01

    We have recently reported significant association of non-polio enteroviruses (NPEVs) with acute and persistent diarrhea (18-21% of total diarrheal cases), and non-diarrheal Increased Frequency of Bowel Movements (IFoBM-ND) (about 29% of the NPEV infections) in children and that the NPEV-associated diarrhea was as significant as rotavirus diarrhea. However, their diarrhea-causing potential is yet to be demonstrated in an animal model system. Since the determination of virus titers by the traditional plaque assay takes 4-7 days, there is a need for development of a rapid method for virus titer determination to facilitate active clinical research on enterovirus-associated diarrhea. The goal of this study is to develop a cell-based rapid detection and enumeration method and to demonstrate the diarrhea-inducing potential of purified and characterized non-polio enteroviruses, which were isolated from diarrheic children. Here we describe generation of monoclonal and polyclonal antibodies against purified strains belonging to different serotypes, and development of an enzyme-linked immuno focus assay (ELIFA) for detection and enumeration of live NPEV particles in clinical and purified virus samples, and a newborn mouse model for NPEV diarrhea. Plaque-purified NPVEs, belonging to different serotypes, isolated from children with diarrhea, were grown in cell culture and purified by isopycnic CsCl density gradient centrifugation. By ELIFA, NPEVs could be detected and enumerated within 12h post-infection. Our results demonstrated that Coxsackievirus B1 (CVB1) and CVB5 strains, isolated from diarrheic children, induced severe diarrhea in orally-inoculated 9-12 day-old mouse pups, fulfilling Koch's postulates. The methods described here would facilitate studies on NPEV-associated gastrointestinal disease. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Growing Up of Autonomous Agents: an Emergent Phenomenon

    Science.gov (United States)

    Morgavi, Giovanna; Marconi, Lucia

    2008-10-01

    A fundamental research challenge is the design of robust artifacts that are capable of operating under changing environments and noisy input, and yet exhibit the desired behavior and response time. These systems should be able to adapt and learn how to react to unforeseen scenarios as well as to display properties comparable to biological entities. The turn to nature has brought us many unforeseen great concepts. Biological systems are able to handle many of these challenges with an elegance and efficiency still far beyond current human artifacts. A living artifact grows up when its capabilities, abilities/knowledge, shift to a further level of complexity, i.e. the complexity rank of its internal capabilities performs a step forward. In the attempt to define an architecture for autonomous growing up agents [1]. We conducted an experiment on the abstraction process in children as natural parts of a cognitive system. We found that linguistic growing up involve a number of different trial processes. We identified a fixed number of distinct paths that were crossed by children. Once a given interpretation paths was discovered useless, they tried to follow another path, until the new meaning was emerging. This study generates suggestion about the evolutionary conditions conducive to the emergence of growing up in robots and provides guidelines for designing artificial evolutionary systems displaying spontaneous adaptation abilities. The importance of multi-sensor perception, motivation and emotional drives are underlined and, above all, the growing up insights shows similarities to emergent self-organized behaviors.

  20. Rapid and sensitive method for quantification of gestodene in human plasma as the oxime derivative by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application to bioequivalence study.

    Science.gov (United States)

    Saxena, Ashish; Gupta, Arun; Kasibhatta, Ravisekhar; Bob, Manoj; Kumar, V Praveen; Purwar, Bipin

    2014-01-15

    A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the estimation of gestodene in human plasma. Gestodene was extracted from human plasma by using solid-phase extraction technique. Gestodene D6 was used as the internal standard. An Acquity HSS-T3 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The mass transition ion-pair was followed as m/z 326.2→124.1 for gestodene and m/z 332.3→129.1 for gestodene D6. The method involves a solid phase extraction from plasma, rapid derivatization with hydroxylamine to form oxime, simple gradient chromatographic conditions and mass spectrometric detection that enables detection at sub-picogram levels. The proposed method has been validated for a linear range of 50-11957pg/ml with a correlation coefficient≥0.9994. The intra-run and inter-run precision and accuracy were within 10%. The overall recoveries for gestodene and gestodene D6 were 62.02% and 67.57% respectively. The total run time was 4.0min. The developed method was applied for the determination of the pharmacokinetic parameters of gestodene following a single oral administration of a 2×0.06mg gestodene tablets in 10 healthy female volunteers. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Rapid liquid chromatography/tandem mass spectrometer (LCMS) method for clozapine and its metabolite N-desmethyl clozapine (norclozapine) in human serum.

    Science.gov (United States)

    Rao, L V; Snyder, M L; Vallaro, G M

    2009-01-01

    Clozapine is indicated for the treatment of schizophrenia and related psychotic disorders. Several methods have been developed for monitoring Clozapine levels; however, they possess limited specificity and are often laborious. This study describes a simple liquid chromatography/tandem mass spectrometer (LCMS) method in human serum. The ion transitions monitored were m/z 327, 270, 296 for Clozapine, m/z 313, 192, 227 for Norclozapine and m/z 328, 271 for Loxapine. The assay is linear (25-1000 ng/ml) and showed a good correlation (r=0.98) within the analytical range of 79-1210 ng/ml in human serum. This assay is highly specific and sensitive for the simultaneous measurements of Clozapine and Norclozapine. The simplification of this assay makes it ideal for high throughput analyses of the patient samples in a routine clinical laboratory staffed with general medical technologists.

  2. The Silk-protein Sericin Induces Rapid Melanization of Cultured Primary Human Retinal Pigment Epithelial Cells by Activating the NF-κB Pathway

    OpenAIRE

    J. R. Eidet; Reppe, S.; Pasovic, L.; Olstad, O K; Lyberg, T; Khan, A. Z.; Fostad, I. G.; D. F. Chen; Utheim, T. P.

    2016-01-01

    Restoration of the retinal pigment epithelial (RPE) cells to prevent further loss of vision in patients with age-related macular degeneration represents a promising novel treatment modality. Development of RPE transplants, however, requires up to 3 months of cell differentiation. We explored whether the silk protein sericin can induce maturation of primary human retinal pigment epithelial (hRPE) cells. Microarray analysis demonstrated that sericin up-regulated RPE-associated transcripts (RPE6...

  3. Magnetically controlled growing rods for scoliosis surgery.

    Science.gov (United States)

    Metkar, Umesh; Kurra, Swamy; Quinzi, David; Albanese, Stephen; Lavelle, William F

    2017-02-01

    Early onset scoliosis can be both a disfiguring as well as a life threatening condition. When more conservative treatments fail, pediatric spinal surgeons are forced to consider operative interventions. Traditionally, these interventions have involved the insertion of a variety of implants into the patient with a limited number of anchor points controlling the spine. In the past, these pediatric patients have had multiple surgeries for elective lengthening of these devices to facilitate their growth while attempting to control the scoliosis. These patients often experience a physical and emotional toll from their multiple repeated surgeries. Growing spine techniques have also had a noted high complication rate due to implant dislodgement and infections. Recently, the development of non-invasively, self-lengthening growing rods has occurred. These devices have the potential to allow for the devices to be lengthened magnetically in a conscious patient in the surgeon's office. Areas covered: This review summarized previously published articles in the English literature using a key word search in PubMed for: 'magnetically controlled growing rods', 'Magec rods', 'magnetic growing rods' and 'growing rods'. Expert commentary: Magnetically controlled growing rods have an advantage over growing rods in lengthening the growing spine in the absence of repetitive surgeries.

  4. (GrOW) Frequently Asked Questions

    International Development Research Centre (IDRC) Digital Library (Canada)

    Alejandra

    How many applications will be shortlisted at the end of the first stage? GrOW envisages that up to 4 selected teams will be asked to submit a full research proposal for review and funding consideration. How is the review process conducted? Upon receiving complete applications, a review process begins by the GrOW team, ...

  5. Growing wealth and growing pains: child and adolescent psychiatry in Hong Kong, Malaysia and Singapore.

    Science.gov (United States)

    Tan, Susan; Fung, Daniel; Hung, Se-Fong; Rey, Joseph

    2008-06-01

    Several Asian regions have undergone a dramatic transformation, some becoming very affluent. This paper aims to ascertain how countries that are becoming wealthy have dealt with child and adolescent mental health issues. Population health status, child and adolescent mental health services, child psychiatry training, the number of child psychiatrists and related matters were examined in Hong Kong, Malaysia and Singapore. Hong Kong, Malaysia and Singapore are ethnically, religiously, socially and politically very different. In spite of considerable wealth and a growing recognition that mental health problems in the young are increasing, they face similar problems--lack of access to treatment due to a dearth of services and a lack of child psychiatrists (2.5, 0.5 and 2.8 per million people, respectively). Because the number of child psychiatrists is so small, their ability to provide services, advocate, train, maintain a professional identity, and deal with future crises is very limited. Other rapidly developing countries can learn from this experience and should take action early to prevent a similar outcome.

  6. Human and Climate Amplified Rapid River Bluff Erosion: an Application of Terrestrial-Based Structure-from-Motion and Time-Lapse Photogrammetry in the Midwestern USA

    Science.gov (United States)

    Kelly, S. A.; Belmont, P.

    2016-12-01

    The Minnesota River Basin (MRB), south central Minnesota, is an extraordinary natural laboratory, and is currently impaired for excess sediments and nutrients. Tributaries of the Minnesota River have been actively incising and meandering into glacial tills for the last 13,400 years in response to a catastrophic base level drop. It is this geologic history combined with modern agricultural intensification and hydrologic change that provide a unique opportunity to study remarkably rapid bluff erosion using photogrammetry. Here we seek to understand the dominant mechanisms (fluvial toe erosion, freeze-thaw, groundwater sapping, tree throw, positive pore water pressures) and magnitude of bluff erosion using daily time-lapse photographs, repeat photo surveys, Structure-from-Motion (SfM), and calculations of bluff volumetric change through time. We expect for increased precipitation and streamflows in the basin to have accelerated bluff erosion via channel widening and migration. In general, we find that bluff failures occur during or immediately following (1-2 days) heavy rainfall events, especially during spring thaw, and the removal, or transport of the failed material out of the reach coincides with hydrograph flood peaks. Through the combined use of time-lapse and SfM photogrammetry we are able to identify the precipitation and flow events which occur with a frequency and magnitude large enough to effectively dominate bluff erosion. Identifying this `effective discharge' of bluff erosion is regionally important for sediment reduction strategies in the MRB.

  7. Viremic blood donor found by a rapid screening method in a season of high human parvovirus B19 activity in Niterói, Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Sérgio Setúbal

    2004-02-01

    Full Text Available Erythrovirus B19 infection is usually benign but may have serious consequences in patients with hemolytic anemia (transient aplastic crisis, immunodeficiency (in whom persistent infection can lead to chronic bone marrow failure with anemia, or who are in the first or second trimester of gestation (spontaneous abortion, hydrops fetalis, and fetal death. Being non-enveloped, B19 resists most inactivation methods and can be transmitted by transfusion. B19 is difficult to cultivate and native virus is usually obtained from viremic blood. As specific antibodies may be absent, and there is no reliable immunological method for antigen detection, hybridization or polymerase chain reaction are needed for detecting viremia. A rapid method, gel hemagglutination (Diamed ID-Parvovirus B19 Antigen Test, can disclose highly viremic donations, whose elimination lessens the viral burden in pooled blood products and may even render them non-infectious. In order to obtain native antigen and to determine the frequency of viremic donors, we applied this test to blood donors in a period of high viral activity in our community. Positive or indeterminate results were re-tested by dot-blot hybridization. We tested 472 donors in 1998 and 831 ones in 1999. One viremic donor was found in 1999. We suggest that in periods of high community viral activity the gel hemagglutination test may be useful in avoiding highly viremic blood being added to plasma pools or directly transfused to patients under risk.

  8. Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

    Science.gov (United States)

    Lindborg, Beth A; Brekke, John H; Vegoe, Amanda L; Ulrich, Connor B; Haider, Kerri T; Subramaniam, Sandhya; Venhuizen, Scott L; Eide, Cindy R; Orchard, Paul J; Chen, Weili; Wang, Qi; Pelaez, Francisco; Scott, Carolyn M; Kokkoli, Efrosini; Keirstead, Susan A; Dutton, James R; Tolar, Jakub; O'Brien, Timothy D

    2016-07-01

    Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. ©AlphaMed Press.

  9. Stability evaluation and sensitive determination of antiviral drug, valacyclovir and its metabolite acyclovir in human plasma by a rapid liquid chromatography-tandem mass spectrometry method.

    Science.gov (United States)

    Yadav, Manish; Upadhyay, Vivek; Singhal, Puran; Goswami, Sailendra; Shrivastav, Pranav S

    2009-03-15

    A simple, sensitive and high throughput liquid chromatography/positive-ion electrospray ionization mass spectrometry (LC-ESI-MS/MS) method has been developed for the simultaneous determination of valacyclovir and acyclovir in human plasma using fluconazole as internal standard (IS). The method involved solid phase extraction of the analytes and IS from 0.5 mL human plasma with no reconstitution and drying steps (direct injection of eluate). The chromatographic separation was achieved on a Gemini C18 analytical column using isocratic mobile phase, consisting of 0.1% formic acid and methanol (30:70 v/v), at a flow-rate of 0.8 mL/min. The precursor-->product ion transition for valacyclovir (m/z 325.2-->152.2), acyclovir (m/z 226.2-->152.2) and IS (m/z 307.1-->220.3) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) mode. The method was validated over the concentration range 5.0-1075 ng/mL and 47.6-10225 ng/mL for valacyclovir and acyclovir respectively. The mean recovery of valacyclovir (92.2%), acyclovir (84.2%) and IS (103.7%) from spiked plasma samples was consistent and reproducible. The bench top stability of valacyclovir and acyclovir was extensively evaluated in buffered and unbuffered plasma. It was successfully applied to a bioequivalence study in 41 healthy human subjects after oral administration of 1000 mg valacyclovir tablet formulation under fasting condition.

  10. Actin filament-associated protein 1 (AFAP-1) is a key mediator in inflammatory signaling-induced rapid attenuation of intrinsic P-gp function in human brain capillary endothelial cells.

    Science.gov (United States)

    Hoshi, Yutaro; Uchida, Yasuo; Tachikawa, Masanori; Ohtsuki, Sumio; Terasaki, Tetsuya

    2017-04-01

    The purpose of this study was to identify regulatory molecule(s) involved in the inflammatory signaling-induced decrease in P-glycoprotein (P-gp) efflux function at the blood-brain barrier (BBB) that may occur in brain diseases. We confirmed that in vivo P-gp efflux activity at the BBB was decreased without any change in P-gp protein expression level in a mouse model of acute inflammation induced by 3 mg/kg lipopolysaccharide. In a human BBB model cell line (human brain capillary endothelial cells; hCMEC/D3), 1-h treatment with 10 ng/mL tumor necrosis factor-α (TNF-α; an inflammatory mediator) rapidly reduced P-gp efflux activity, but had no effect on P-gp protein expression level. To clarify the non-transcriptional mechanism that causes the decrease in intrinsic efflux activity of P-gp in acute inflammation, we applied comprehensive quantitative phosphoproteomics to compare hCMEC/D3 cells treated with TNF-α and vehicle (control). Actin filament-associated protein-1 (AFAP-1), MAPK1, and transcription factor AP-1 (AP-1) were significantly phosphorylated in TNF-α-treated cells, and were selected as candidate proteins. In validation experiments, knockdown of AFAP-1 expression blocked the reduction in P-gp efflux activity by TNF-α treatment, whereas inhibition of MAPK function or knockdown of AP-1 expression did not. Quantitative targeted absolute proteomics revealed that the reduction in P-gp activity by TNF-α did not require any change in P-gp protein expression levels in the plasma membrane. Our results demonstrate that AFAP-1 is a key mediator in the inflammatory signaling-induced, translocation-independent rapid attenuation of P-gp efflux activity in human brain capillary endothelial cells. © 2017 International Society for Neurochemistry.

  11. A rapid ultrasound-assisted dispersive liquid-liquid microextraction followed by ultra-performance liquid chromatography for the simultaneous determination of seven benzodiazepines in human plasma samples.

    Science.gov (United States)

    Fernández, Purificación; González, Cristina; Pena, M Teresa; Carro, Antonia M; Lorenzo, Rosa A

    2013-03-12

    A simple and efficient ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) method has been developed for the determination of seven benzodiazepines (alprazolam, bromazepam, clonazepam, diazepam, lorazepam, lormetazepam and tetrazepam) in human plasma samples. Chloroform and methanol were used as extractant and disperser solvents, respectively. The influence of several variables (e.g., type and volume of dispersant and extraction solvents, pH, ultrasonic time and ionic strength) was carefully evaluated and optimized, using an asymmetric screening design 3(2)4(2)//16. Analysis of extracts was performed by ultra-performance liquid chromatography coupled with photodiode array detection (UPLC-PDA). Under the optimum conditions, two reversed-phases, Shield RP18 and C18 columns were successfully tested, obtaining good linearity in a range of 0.01-5μgmL(-1), with correlation coefficients r>0.996. Quantification limits ranged between 4.3-13.2ngmL(-1) and 4.0-14.8ngmL(-1), were obtained for C18 and Shield RP18 columns, respectively. The optimized method exhibited a good precision level, with relative standard deviation values lower than 8%. The recoveries studied at two spiked levels, ranged from 71 to 102% for all considered compounds. The proposed method was successfully applied to the analysis of seven benzodiazepines in real human plasma samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Rapid determination of rivaroxaban in human urine and serum using colloidal palladium surface-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Cheng, Yu-Han; Chen, Wen-Chi; Chang, Sarah Y

    2015-11-15

    Rivaroxaban is a new anticoagulant drug that has recently been introduced for clinical applications. To ensure optimum efficacy while minimizing the risk of toxicity and other adverse effects, a simple and sensitive analytical procedure for monitoring the concentration of rivaroxaban in biological fluids is required. Rivaroxaban was extracted from aqueous solutions by dispersive liquid-liquid microextraction (DLLME). Detection of rivaroxaban was achieved through surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) using colloidal palladium as the SALDI matrix. The calibration curve for rivaroxaban in aqueous solutions was linear over the concentration range from 5 to 500 nM. The limit of detection (LOD) for rivaroxaban at a signal-to-noise ratio of 3 was 2 nM. With a sample-to-extract volume ratio of 200, the enrichment factors were calculated to be 141. This method was successfully applied for the determination of rivaroxaban in human urine and serum samples. The LODs for rivaroxaban in urine and serum were calculated to be 6 nM and 60 nM, respectively. The analysis speed, together with the ease of operation and high sensitivity, allows SALDI-MS method to be particularly suitable for the high-throughput screening of rivaroxaban levels in human urine and serum samples. Copyright © 2015 John Wiley & Sons, Ltd.

  13. A Rapid and Optimized LC-MS/MS Method for the Simultaneous Extraction and Determination of Sofosbuvir and Ledipasvir in Human Plasma.

    Science.gov (United States)

    Elkady, Ehab F; Aboelwafa, Ahmed A

    2016-09-01

    A new validated bioanalytical method based on LC tandem MS has been developed for the simultaneous extraction and determination of sofosbuvir and ledipasvir in human plasma using antiviral daclatasvir as an internal standard (IS). Liquid-liquid extraction of samples was used for the purification and preconcentration of the analytes from a human plasma matrix. Good and consistent recoveries were obtained, with average extraction recoveries of 91.61 and 88.93% for sofosbuvir and ledipasvir, respectively. The chromatographic separation of the three analytes was achieved within only 2.8 min by an isocratic mobile phase consisting of 10 mM ammonium acetate, which was then adjusted to pH 4.0 by acetic acid-acetonitrile-0.1% methanolic formic acid (12 + 25 + 63, v/v/v) flowing through a C18 Zorbax eclipse plus column (5 μm, 100 × 4.6 mm; Agilent). Multiple reaction monitoring transitions were measured in positive ion mode for sofosbuvir, ledipasvir, and daclatasvir (IS). A detailed validation of the method was performed and the standard curves were found to be linear in the range of 0.5 to 2500 and 5 to 2100 ng/mL for sofosbuvir and ledipasvir, respectively, applying weighted (1/X(2)) linear regression. The developed method was applied to the analysis of the two drugs after a single oral administration of Harvoni 400/90 mg film-coated tablets containing 400 mg sofosbuvir and 90 mg ledipasvir to four healthy volunteers.

  14. CD133-enriched Xeno-Free human embryonic-derived neural stem cells expand rapidly in culture and do not form teratomas in immunodeficient mice

    Directory of Open Access Journals (Sweden)

    Daniel L. Haus

    2014-09-01

    Full Text Available Common methods for the generation of human embryonic-derived neural stem cells (hNSCs result in cells with potentially compromised safety profiles due to maintenance of cells in conditions containing non-human proteins (e.g. in bovine serum or on mouse fibroblast feeders. Additionally, sufficient expansion of resulting hNSCs for scaling out or up in a clinically relevant time frame has proven to be difficult. Here, we report a strategy that produces hNSCs in completely “Xeno-Free” culture conditions. Furthermore, we have enriched the hNSCs for the cell surface marker CD133 via magnetic sorting, which has led to an increase in the expansion rate and neuronal fate specification of the hNSCs in vitro. Critically, we have also confirmed neural lineage specificity upon sorted hNSC transplantation into the immunodeficient NOD-scid mouse brain. The future use or adaptation of these protocols has the potential to better facilitate the advancement of pre-clinical strategies from the bench to the bedside.

  15. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of

  16. Rapid 3D in vivo 1H human lung respiratory imaging at 1.5 T using ultra-fast balanced steady-state free precession.

    Science.gov (United States)

    Pusterla, Orso; Bauman, Grzegorz; Wielpütz, Mark O; Nyilas, Sylvia; Latzin, Philipp; Heussel, Claus P; Bieri, Oliver

    2017-09-01

    To introduce a reproducible, nonenhanced 1H MRI method for rapid in vivo functional assessment of the whole lung at 1.5 Tesla (T). At different respiratory volumes, the pulmonary signal of ultra-fast steady-state free precession (ufSSFP) follows an adapted sponge model, characterized by a respiratory index α. From the model, α reflects local ventilation-related information, is virtually independent from the lung density and thus from the inspiratory phase and breathing amplitude. Respiratory α-mapping is evaluated for healthy volunteers and patients with obstructive lung disease from a set of five consecutive 3D ultra-fast steady-state free precession (ufSSFP) scans performed in breath-hold and at different inspiratory volumes. For the patients, α-maps were compared with CT, dynamic contrast-enhanced MRI (DCE-MRI), and Fourier decomposition (FD). In healthy volunteers, respiratory α-maps showed good reproducibility and were homogeneous on iso-gravitational planes, but showed a gravity-dependent respiratory gradient. In patients with obstructive pulmonary disease, the functional impairment observed in respiratory α-maps was associated with emphysematous regions present on CT images, perfusion defects observable on DCE-MRI, and impairments visualized on FD ventilation and perfusion maps. Respiratory α-mapping derived from multivolumetric ufSSFP provides insights into functional lung impairment and may serve as a reproducible and normative measure for clinical studies. Magn Reson Med 78:1059-1069, 2017. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

  17. Rapid, progressive neuropathic arthropathy of the hip in a patient co-infected with human immunodeficiency virus, hepatitis C virus and tertiary syphilis: case report

    Science.gov (United States)

    2011-01-01

    Background Syphilis is a chronic infection that is classified into three stages. In its tertiary stage, syphilis spreads to the brain, heart and other organs; the lesions may involve the skin, mucous membranes and bones. Neuropathic arthropathy associated with tertiary syphilis has rarely been described in Europe and its association with HIV-HCV co-infection has not been reported so far. This article reports the case of a man with tertiary syphilis presenting with rapidly evolving neuropathic arthropathy of the hip and extensive bone destruction. Case presentation On initial presentation, the patient complained of progressively worsening left-sided coxalgia without localized or generalized inflammation. The patient reported to have no history of previous infections, trauma or cancer. Plain x-ray films of the left coxofemoral joint showed marked degeneration with necrosis of the proximal epiphysis of femur and morphological alterations of the acetabulum without protrusion. Primary coxarthrosis was diagnosed and hip arthroplasty was offered, but the patient declined treatment. Three months later, the patient presented a marked deterioration of his general condition. He disclosed that he was seropositive for HCV and HIV, as confirmed by serology. Syphilis serology testing was also positive. A Girdlestone's procedure was performed and samples were collected for routine cultures for bacteria and acid fast bacilli, all resulting negative. Although histological findings were inconclusive, confirmed positive serology for syphilis associated with progressive arthropathy was strongly suggestive of tertiary syphilis, probably exacerbated by HIV-HCV co-infection. The patient partially recovered the ability to walk. Conclusions Due to the resurgence of syphilis, this disease should be considered as a possible cause of neuropathic arthropathy when other infectious causes have been ruled out, particularly in patients with HIV and/or HCV co-infection. PMID:21645338

  18. Rapid, progressive neuropathic arthropathy of the hip in a patient co-infected with human immunodeficiency virus, hepatitis C virus and tertiary syphilis: case report

    Directory of Open Access Journals (Sweden)

    Pasqualini Marco

    2011-06-01

    Full Text Available Abstract Background Syphilis is a chronic infection that is classified into three stages. In its tertiary stage, syphilis spreads to the brain, heart and other organs; the lesions may involve the skin, mucous membranes and bones. Neuropathic arthropathy associated with tertiary syphilis has rarely been described in Europe and its association with HIV-HCV co-infection has not been reported so far. This article reports the case of a man with tertiary syphilis presenting with rapidly evolving neuropathic arthropathy of the hip and extensive bone destruction. Case presentation On initial presentation, the patient complained of progressively worsening left-sided coxalgia without localized or generalized inflammation. The patient reported to have no history of previous infections, trauma or cancer. Plain x-ray films of the left coxofemoral joint showed marked degeneration with necrosis of the proximal epiphysis of femur and morphological alterations of the acetabulum without protrusion. Primary coxarthrosis was diagnosed and hip arthroplasty was offered, but the patient declined treatment. Three months later, the patient presented a marked deterioration of his general condition. He disclosed