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Sample records for rapidly growing clones

  1. Management of Infections with Rapidly Growing

    Directory of Open Access Journals (Sweden)

    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.

  2. Water use by Eucalypt Clones Growing at Kongowe, Kibaha, Tanzania

    African Journals Online (AJOL)

    A study was carried out between April and October 2014 to investigate the amount of water used by Eucalypt clones grown at Kongowe, Tanzania. Sap flow sensors using heat pulse velocity were deployed across GC 167, GC 15 and GC 940 in wet and dry seasons. Data on water use were analysed using sap flow software ...

  3. A rapid cloning method employing orthogonal end protection

    NARCIS (Netherlands)

    Jakobi, A.J.|info:eu-repo/dai/nl/311489621; Huizinga, E.G.|info:eu-repo/dai/nl/12314969X

    2012-01-01

    We describe a novel in vitro cloning strategy that combines standard tools in molecular biology with a basic protecting group concept to create a versatile framework for the rapid and seamless assembly of modular DNA building blocks into functional open reading frames. Analogous to chemical

  4. 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

  5. Environmental and Social Programmes and Rapidly Growing Retailers

    Directory of Open Access Journals (Sweden)

    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.

  6. A rapid cloning method employing orthogonal end protection.

    Directory of Open Access Journals (Sweden)

    Arjen J Jakobi

    Full Text Available We describe a novel in vitro cloning strategy that combines standard tools in molecular biology with a basic protecting group concept to create a versatile framework for the rapid and seamless assembly of modular DNA building blocks into functional open reading frames. Analogous to chemical synthesis strategies, our assembly design yields idempotent composite synthons amenable to iterative and recursive split-and-pool reaction cycles. As an example, we illustrate the simplicity, versatility and efficiency of the approach by constructing an open reading frame composed of tandem arrays of a human fibronectin type III (FNIII domain and the von Willebrand Factor A2 domain (VWFA2, as well as chimeric (FNIII(n-VWFA2-(FNIII(n constructs. Although we primarily designed this strategy to accelerate assembly of repetitive constructs for single-molecule force spectroscopy, we anticipate that this approach is equally applicable to the reconstitution and modification of complex modular sequences including structural and functional analysis of multi-domain proteins, synthetic biology or the modular construction of episomal vectors.

  7. The Commercial Profitability of Growing Hybrid Eucalyptus Clones in The Coast Province, Kenya

    Directory of Open Access Journals (Sweden)

    Balozi Bekuta Kirongo

    2014-04-01

    Full Text Available Due to the current high demand for timber, fuelwood, and building poles and the realization that tree growing may pay dividends in the short and long term, many farmers are planting trees on their farms. Farmers are increasingly planting eucalyptus partly due to the fast growth rates of the hybrid clones as well as the opportunity to earn money within a short time. In this paper we report on the profitability of growing eucalyptus hybrid clones in the coastal region, Kenya. Tree growth and cost data was sourced from farmers in Malindi, Kilifi, and Msambweni. Market information was sourced from hardwares in North and South Coast while tree growth models were used to provide average tree sizes at various ages. Results showed that a farmer could make a net income of upto Kshs.500,000.00 (USD6,250 in 5 years. Farmers in the South Coast (Kwale and Msambweni spent more on transport than their counterparts in the North Coast (near Gede-KEFRI. This, added to the fact that trees in the South Coast (Msambweni grew less compared to those in North Coast meant that farmers in the south made less profits. Keywords: profitability, eucalyptus on-farm tree growing, renewable energy, aforestation

  8. Comparative Genomic Analysis of Rapid Evolution of an Extreme-Drug-Resistant Acinetobacter baumannii Clone

    Science.gov (United States)

    Tan, Sean Yang-Yi; Chua, Song Lin; Liu, Yang; Høiby, Niels; Andersen, Leif Percival; Givskov, Michael; Song, Zhijun; Yang, Liang

    2013-01-01

    The emergence of extreme-drug-resistant (EDR) bacterial strains in hospital and nonhospital clinical settings is a big and growing public health threat. Understanding the antibiotic resistance mechanisms at the genomic levels can facilitate the development of next-generation agents. Here, comparative genomics has been employed to analyze the rapid evolution of an EDR Acinetobacter baumannii clone from the intensive care unit (ICU) of Rigshospitalet at Copenhagen. Two resistant A. baumannii strains, 48055 and 53264, were sequentially isolated from two individuals who had been admitted to ICU within a 1-month interval. Multilocus sequence typing indicates that these two isolates belonged to ST208. The A. baumannii 53264 strain gained colistin resistance compared with the 48055 strain and became an EDR strain. Genome sequencing indicates that A. baumannii 53264 and 48055 have almost identical genomes—61 single-nucleotide polymorphisms (SNPs) were found between them. The A. baumannii 53264 strain was assembled into 130 contigs, with a total length of 3,976,592 bp with 38.93% GC content. The A. baumannii 48055 strain was assembled into 135 contigs, with a total length of 4,049,562 bp with 39.00% GC content. Genome comparisons showed that this A. baumannii clone is classified as an International clone II strain and has 94% synteny with the A. baumannii ACICU strain. The ResFinder server identified a total of 14 antibiotic resistance genes in the A. baumannii clone. Proteomic analyses revealed that a putative porin protein was down-regulated when A. baumannii 53264 was exposed to antimicrobials, which may reduce the entry of antibiotics into the bacterial cell. PMID:23538992

  9. The Commercial Profitability of Growing Hybrid Eucalyptus Clones in The Coast Province, Kenya

    Directory of Open Access Journals (Sweden)

    Balozi Bekuta Kirongo

    2014-04-01

    Full Text Available Due to the current high demand for timber, fuelwood, and building poles and the realization that tree growing may pay dividends in the short and long term, many farmers are planting trees on their farms. Farmers are increasingly planting eucalyptus partly due to the fast growth rates of the hybrid clones as well as the opportunity to earn money within a short time. In this paper we report on the profitability of growing eucalyptus hybrid clones in the coastal region, Kenya. Tree growth and cost data was sourced from farmers in Malindi, Kilifi, and Msambweni. Market information was sourced from hardwares in North and South Coast while tree growth models were used to provide average tree sizes at various ages. Results showed that a farmer could make a net income of upto Kshs.500,000.00 (USD6,250 in 5 years. Farmers in the South Coast (Kwale and Msambweni spent more on transport than their counterparts in the North Coast (near Gede-KEFRI. This, added to the fact that trees in the South Coast (Msambweni grew less compared to those in North Coast meant that farmers in the south made less profits.

  10. Cloning

    Science.gov (United States)

    Cloning describes the processes used to create an exact genetic replica of another cell, tissue or organism. ... named Dolly. There are three different types of cloning: Gene cloning, which creates copies of genes or ...

  11. 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

  12. Rapid cloning and bioinformatic analysis of spinach Y chromosome ...

    Indian Academy of Sciences (India)

    The female spinach genome was taken as blocker and cDNA library specifically expressed in Y chromosome was constructed. Moreover, expressed sequence tag (EST) sequences in cDNA library were cloned, sequenced and bioinformatics was analysed. There were 63 valid EST sequences obtained in this study.

  13. Rapid cloning and bioinformatic analysis of spinach Y chromosome ...

    Indian Academy of Sciences (India)

    Jun Gao and Long-Dou Lu. J. Genet. 94, 705–713. Table 1. List of part of spinach Y chromosome-specific ESTs clones by dot blot and their size and homology to the sequences available at the GenBank database by BLASTx software.

  14. 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.

  15. Rapid customised operon assembly by yeast recombinational cloning.

    Science.gov (United States)

    Liu, Michael A; Kenyon, Johanna J; Lee, Jason; Reeves, Peter R

    2017-06-01

    We have developed a system called the Operon Assembly Protocol (OAP), which takes advantage of the homologous recombination DNA repair pathway in Saccharomyces cerevisiae to assemble full-length operons from a series of overlapping PCR products into a specially engineered yeast-Escherichia coli shuttle vector. This flexible, streamlined system can be used to assemble several operon clones simultaneously, and each clone can be expressed in the same E. coli tester strain to facilitate direct functional comparisons. We demonstrated the utility of the OAP by assembling and expressing a series of E. coli O1A O-antigen gene cluster clones containing various gene deletions or replacements. We then used these constructs to assess the substrate preferences of several Wzx flippases, which are responsible for translocation of oligosaccharide repeat units (O units) across the inner membrane during O-antigen biosynthesis. We were able to identify several O unit structural features that appear to be important determinants of Wzx substrate preference. The OAP system should be broadly applicable for the genetic manipulation of any bacterial operon and can be modified for use in other host species. It could also have potential uses in fields such as glycoengineering.

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

    Science.gov (United States)

    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.

  17. Comparative Genomic Analysis of Rapid Evolution of an Extreme-Drug-Resistant Acinetobacter baumannii Clone

    DEFF Research Database (Denmark)

    Tan, Sean Yang-Yi; Chua, Song Lin; Liu, Yang

    2013-01-01

    , comparative genomics has been employed to analyze the rapid evolution of an EDR Acinetobacter baumannii clone from the intensive care unit (ICU) of Rigshospitalet at Copenhagen. Two resistant A. baumannii strains, 48055 and 53264, were sequentially isolated from two individuals who had been admitted to ICU...

  18. Cloning

    Science.gov (United States)

    ... such as the liver, brain and heart. Other consequences include premature aging and problems with the immune system. Another potential ... cell dies. This is part of the natural aging process that seems to happen in all cell types. As a consequence, clones created from a cell taken from an ...

  19. 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...

  20. Rapid cloning and purification of proteins: gateway vectors for protein purification by self-cleaving tags.

    Science.gov (United States)

    Gillies, Alison R; Hsii, Judy F; Oak, Seachol; Wood, David W

    2008-10-01

    We have combined Invitrogen's Gateway cloning technology with self-cleaving purification tags to generate a new system for rapid production of recombinant protein products. To accomplish this, we engineered our previously reported DeltaI-CM cleaving intein to include a Gateway cloning recognition sequence, and demonstrated that the resulting Gateway-competent intein is unaffected. This intein can therefore be used in several previously reported purification methods, while at the same time being compatible with Gateway cloning. We have incorporated this intein into a set of Gateway vectors, which include self-cleaving elastin-like polypeptide (ELP), chitin binding domain (CBD), phasin (polyhydroxybutyrate-binding), or maltose binding domain (MBD) tags. These vectors were verified by Gateway cloning of TEM-1 beta-lactamase and Escherichia coli catalase genes, and the expressed target proteins were purified using the four methods encoded on the vectors. The purification methods were unaffected by replacing the DeltaI-CM intein with the Gateway intein. It was observed that some purification methods were more appropriate for each target than others, suggesting utility of this technology for rapid process identification and optimization. The modular design of the Gateway system and intein purification method suggests that any tag and promoter can be trivially added to this system for the development of additional expression vectors. This technology could greatly facilitate process optimization, allowing several targets and methods to be tested in a high-throughput manner.

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

    Science.gov (United States)

    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.

  2. 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, ...

  3. Rapid restriction enzyme-free cloning of PCR products: a high-throughput method applicable for library construction.

    Directory of Open Access Journals (Sweden)

    Vijay K Chaudhary

    Full Text Available Herein, we describe a novel cloning strategy for PCR-amplified DNA which employs the type IIs restriction endonuclease BsaI to create a linearized vector with four base-long 5'-overhangs, and T4 DNA polymerase treatment of the insert in presence of a single dNTP to create vector-compatible four base-long overhangs. Notably, the insert preparation does not require any restriction enzyme treatment. The BsaI sites in the vector are oriented in such a manner that upon digestion with BsaI, a stuffer sequence along with both BsaI recognition sequences is removed. The sequence of the four base-long overhangs produced by BsaI cleavage were designed to be non-palindromic, non-compatible to each other. Therefore, only ligation of an insert carrying compatible ends allows directional cloning of the insert to the vector to generate a recombinant without recreating the BsaI sites. We also developed rapid protocols for insert preparation and cloning, by which the entire process from PCR to transformation can be completed in 6-8 h and DNA fragments ranging in size from 200 to 2200 bp can be cloned with equal efficiencies. One protocol uses a single tube for insert preparation if amplification is performed using polymerases with low 3'-exonuclease activity. The other protocol is compatible with any thermostable polymerase, including those with high 3'-exonuclease activity, and does not significantly increase the time required for cloning. The suitability of this method for high-throughput cloning was demonstrated by cloning batches of 24 PCR products with nearly 100% efficiency. The cloning strategy is also suitable for high efficiency cloning and was used to construct large libraries comprising more than 108 clones/µg vector. Additionally, based on this strategy, a variety of vectors were constructed for the expression of proteins in E. coli, enabling large number of different clones to be rapidly generated.

  4. A bacterial two-hybrid system that utilizes Gateway cloning for rapid screening of protein-protein interactions.

    Science.gov (United States)

    Karna, S L Rajasekhar; Zogaj, Xhavit; Barker, Jeffrey R; Seshu, Janakiram; Dove, Simon L; Klose, Karl E

    2010-11-01

    Comprehensive clone sets representing the entire genome now exist for a large number of organisms. The Gateway entry clone sets are a particularly useful means to study gene function, given the ease of introduction into any Gateway-suitable destination vector. We have adapted a bacterial two-hybrid system for use with Gateway entry clone sets, such that potential interactions between proteins encoded within these clone sets can be determined by new destination vectors. We show that utilizing the Gateway clone sets for Francisella tularensis and Vibrio cholerae, known interactions between F. tularensis IglA and IglB and V. cholerae VipA and VipB could be confirmed with these destination vectors. Moreover, the introduction of unique tags into each vector allowed for visualization of the expressed hybrid proteins via Western immunoblot. This Gateway-suitable bacterial two-hybrid system provides a new tool for rapid screening of protein-protein interactions.

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

    Science.gov (United States)

    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).

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

    Directory of Open Access Journals (Sweden)

    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.

  7. 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.

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

    Science.gov (United States)

    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.

  9. Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest

    Directory of Open Access Journals (Sweden)

    Herminia Yohko KANAMURA

    1997-11-01

    Full Text Available A S. mansoni adult worm cDNA expression library was screened with sera from baboons in a early phase after infection. The clones that were positive with the early infection sera were examined for reactivity with pre-infection sera and heterologous infection sera. In order to discriminate a positive antibody reaction from the reactivity due to residual anti-E. coli antibodies, an unrelated cDNA clone was plated with the positive clone. The unrelated clone provided the negative background and the contrast necessary to discern a positive antibody reaction. In this way, we were able to eliminate selected clones that were positive with the pre-infection sera or heterologous infection sera. This characterization of the expression library clones enabled us to quickly target only clones with the desired pattern of antibody reactivity for sequencing, subcloning, and expressingUma biblioteca de expressão de cDNA de verme adulto de S. mansoni foi selecionada utilizando-se soros de macacos babuínos em fase inicial da infecção. Os clones que reagiram positivamente com os soros de infecção recente foram examinados quanto à reatividade contra soros normais e soros de infecção heteróloga. Com a finalidade de se conseguir melhor discriminação entre reatividade positiva com o anticorpo específico e aquela devido aos anticorpos anti-E. coli residuais, um clone de cDNA não relacionado ao S. mansoni foi plaqueado em mistura com o clone positivo. O plano de fundo negativo proporcionado pelo clone não relacionado forneceu o contraste necessário para discriminar a reação positiva específica. Esta metodologia de caracterização dos clones de expressão permite alcançar rapidamente os clones que apresentam o perfil de reatividade desejada, para posterior sequenciamento, subclonagem e expressão

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

    Science.gov (United States)

    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.

  11. 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

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

    Science.gov (United States)

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  16. 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.

  17. 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.

  18. 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.

  19. 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.)

  20. A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.

    Science.gov (United States)

    Fowler, Daniel K; Stewart, Scott; Seredick, Steve; Eisen, Judith S; Stankunas, Kryn; Washbourne, Philip

    2016-01-01

    Recombination-based cloning is a quick and efficient way to generate expression vectors. Recent advancements have provided powerful recombinant DNA methods for molecular manipulations. Here, we describe a novel collection of three-fragment MultiSite Gateway cloning system-compatible vectors providing expanded molecular tools for vertebrate research. The components of this toolkit encompass a broad range of uses such as fluorescent imaging, dual gene expression, RNA interference, tandem affinity purification, chemically-inducible dimerization and lentiviral production. We demonstrate examples highlighting the utility of this toolkit for producing multi-component vertebrate expression vectors with diverse primary research applications. The vectors presented here are compatible with other Gateway toolkits and collections, facilitating the rapid generation of a broad range of innovative DNA constructs for biological research.

  1. Rapid approach for cloning bacterial single-genes directly from soils ...

    African Journals Online (AJOL)

    Obtaining functional genes of bacteria from environmental samples usually depends on library-based approach which is not favored as its large amount of work with small possibility of positive clones. A kind of bacterial single-gene encoding glutamine synthetase (GS) was selected as example to detect the efficiency of ...

  2. 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.

  3. 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.

  4. 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...

  5. An Improved Homologous Recombination Method for Rapid Cloning of the Antibody Heavy Chain Gene and Its Comparison with the Homologous Recombination and Traditional Cloning Methods

    Directory of Open Access Journals (Sweden)

    Masoumeh Hajirezaei

    2015-10-01

    Full Text Available Background: The homologous recombination (HR is one of the conventional cloning methods for the production of recombinant DNA. It is a quick method for in vivo DNA cloning without using the high cost restriction enzymes. A few modifications in the cloning procedure can increase the efficiency of this method.Materials and Methods: In this study, effect of heating on the rate of the IgG1 heavy chain gene cloning was investigated in the HR method and then it was compared with HR method without heating and traditional cloning method. For doing this comparison, three cloning methods including HR, HR with the heat treatment, and traditional cloning were used to clone the human IgG1 heavy chain into the pcDNA3.1(+ plasmid.Results: The results showed that adding heat in the HR method converts insert and vector from the double strand DNA to the single strand DNA. This allows them to copulate with each other better and faster than the two other methods. The heat addition also helps the cell enzyme system for a faster and easier recombination and moreover it increases the cloning efficiency of the HR method in case of in vitro processing.Conclusion: The results showed that giving heat in the HR method increases cloning rate 7.5% and this increase reaches 10% in comparison with the traditional method. 

  6. 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

  7. 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

  8. 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

  9. [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

  10. Cloning of novel rice blast resistance genes from two rapidly evolving NBS-LRR gene families in rice.

    Science.gov (United States)

    Guo, Changjiang; Sun, Xiaoguang; Chen, Xiao; Yang, Sihai; Li, Jing; Wang, Long; Zhang, Xiaohui

    2016-01-01

    Most rice blast resistance genes (R-genes) encode proteins with nucleotide-binding site (NBS) and leucine-rich repeat (LRR) domains. Our previous study has shown that more rice blast R-genes can be cloned in rapidly evolving NBS-LRR gene families. In the present study, two rapidly evolving R-gene families in rice were selected for cloning a subset of genes from their paralogs in three resistant rice lines. A total of eight functional blast R-genes were identified among nine NBS-LRR genes, and some of these showed resistance to three or more blast strains. Evolutionary analysis indicated that high nucleotide diversity of coding regions served as important parameters in the determination of gene resistance. We also observed that amino-acid variants (nonsynonymous mutations, insertions, or deletions) in essential motifs of the NBS domain contribute to the blast resistance capacity of NBS-LRR genes. These results suggested that the NBS regions might also play an important role in resistance specificity determination. On the other hand, different splicing patterns of introns were commonly observed in R-genes. The results of the present study contribute to improving the effectiveness of R-gene identification by using evolutionary analysis method and acquisition of novel blast resistance genes.

  11. Raman spectroscopy for rapid discrimination of Staphylococcus epidermidis clones related to medical device-associated infections

    Science.gov (United States)

    Samek, O.; Telle, H. H.; Harris, L. G.; Bloomfield, M.; Mack, D.

    2008-06-01

    We report on the potential application of Raman spectroscopy for the fast typing of Staphylococcus epidermidis (S. epidermidis) strains related to medical device-associated infections. In this study bacterial colonies were directly probed on culture plates and Raman spectra were recorded from volumes containing approximately 10 bacteria. The spectra contain information on the molecular composition of the whole bacteria, such as fatty acids, carbohydrates, proteins and nucleic acids, DNA as well as RNA. We demonstrate the potential to discriminate different S. epidermidis clones, even after only short Raman exposure/collection times.

  12. Heterogeneity of DNA content and expression of cell cycle genes in axenically growing Entamoeba histolytica HM1:IMSS clone A.

    Science.gov (United States)

    Gangopadhyay, S S; Ray, S S; Kennady, K; Pande, G; Lohia, A

    1997-12-01

    The cell division cycle of Entamoeba histolytica was studied using multi-parametric flow cytometry in asynchronous and partially synchronised cells. Dynamic changes in the DNA synthesis and DNA content of axenically growing trophozoites were observed by using 5-bromo-2'-deoxyuridine (BrdU) uptake and DNA specific fluorochromes. It was observed that DNA synthesis in these cells continues beyond the typical S-phase stop point when DNA duplication is complete. Asynchronously growing E. histolytica cells could be synchronised by serum starvation followed by serum re-addition. BrdU incorporation in synchronised cells showed that cell synchrony is maintained for at least one generation time, in which the G1 phase lasts for 2-3 h and the S-phase lasts for 5-6 h. Analysis of our results revealed that E. histolytica trophozoites, growing in axenic medium, are made up of a heterogenous population of euploid and polyploid cells. The number of polyploid cells increases with age of the cells in culture. Expression of putative cell cycle and signal transduction markers was studied using specific antibodies and changes in their expression levels have been correlated with changes in the DNA content. Based upon our results we could identify G1, S and G2 phases of the cell cycle of E. histolytica and also predict the mechanism underlying the generation of polyploidy in these cells, which may have significant effects on its biology and pathogenesis.

  13. Rapid Construction of Complex Plant RNA Virus Infectious cDNA Clones for Agroinfection Using a Yeast-E. coli-Agrobacterium Shuttle Vector.

    Science.gov (United States)

    Sun, Kai; Zhao, Danyang; Liu, Yong; Huang, Changjun; Zhang, Wei; Li, Zhenghe

    2017-11-07

    The availability of infectious full-length clone is indispensable for reverse genetics studies of virus biology, pathology and construction of viral vectors. However, for RNA viruses with large genome sizes or those exhibiting inherent cloning difficulties, procedure to generate biologically active circular DNA (cDNA) clones can be time-consuming or technically challenging. Here we have constructed a yeast- Escherichia coli - Agrobacterium shuttle vector that enables highly efficient homologous recombination in yeast for assembly of Agrobacterium compatible plant virus clones. Using this vector, we show that infectious cDNA clones of a plant negative-stranded RNA virus, sonchus yellow net rhabdovirus, can be rapidly assembled. In addition, one-step assembly of infectious clones of potato virus Y in yeast, either with or without intron, was readily achieved from as many as eight overlapping DNA fragments. More importantly, the recovered yeast plasmids can be transformed directly into Agrobacterium for inoculation, thereby obviating the E. coli cloning steps and associated toxicity issues. This method is rapid, highly efficient and cost-effective and should be readily applicable to a broad range of plant viruses.

  14. Rapid Construction of Complex Plant RNA Virus Infectious cDNA Clones for Agroinfection Using a Yeast-E. coli-Agrobacterium Shuttle Vector

    Directory of Open Access Journals (Sweden)

    Kai Sun

    2017-11-01

    Full Text Available The availability of infectious full-length clone is indispensable for reverse genetics studies of virus biology, pathology and construction of viral vectors. However, for RNA viruses with large genome sizes or those exhibiting inherent cloning difficulties, procedure to generate biologically active complementary DNA (cDNA clones can be time-consuming or technically challenging. Here we have constructed a yeast-Escherichia coli-Agrobacterium shuttle vector that enables highly efficient homologous recombination in yeast for assembly of Agrobacterium compatible plant virus clones. Using this vector, we show that infectious cDNA clones of a plant negative-stranded RNA virus, sonchus yellow net rhabdovirus, can be rapidly assembled. In addition, one-step assembly of infectious clones of potato virus Y in yeast, either with or without intron, was readily achieved from as many as eight overlapping DNA fragments. More importantly, the recovered yeast plasmids can be transformed directly into Agrobacterium for inoculation, thereby obviating the E. coli cloning steps and associated toxicity issues. This method is rapid, highly efficient and cost-effective and should be readily applicable to a broad range of plant viruses.

  15. 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.

  16. 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...

  17. 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.

  18. 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.

  19. Effect of Low pH and Aluminum Toxicity on the Photosynthetic Characteristics of Different Fast-Growing Eucalyptus Vegetatively Propagated Clones

    Science.gov (United States)

    Yang, Mei; Tan, Ling; Xu, Yuanyuan; Zhao, Yihui; Cheng, Fei; Ye, Shaoming; Jiang, Weixin

    2015-01-01

    Knowing how acid soils and aluminum in soils may limit the growth of Eucalyptus trees in plantations is important because these plantations grow in many tropical and subtropical regions. Seedlings of four vegetatively propagated Eucalyptus clones, E. grandis × E. urophylla ‘GLGU9’(G9), E. grandis × E. urophylla ‘GLGU12’ (G12), E. urophylla × E. camaldulensis ‘GLUC3’ (G3) and E. urophylla ‘GLU4’(G4), were subjected to liquid culture with Hoagland nutrient solution for 40 days, then treated with four different treatments of acid and aluminum for 1 day. The four treatments used either pH 3.0 or 4.0 with or without added aluminum (4.4 mM) in all possible combinations; a control used no added aluminum at pH 4.8. Subsequently, the photosynthetic parameters and morphology of leaves from eucalypt seedlings were determined and observed. The results showed that the tested chlorophyll content, net photosynthetic rate, transpiration rate and water use efficiency were apparently inhibited by aluminum. Under uniform Al concentration (4.4 mM), the Al-induced limitation to photosynthetic parameters increased with pH, indicating acid stimulation to Al toxicity. Among all treatments, the most significant reduction was found in the combination of pH 3.0 and 4.4 mM Al. The photosynthetic and transpiration rates showed similar trends with G9 > G12 > G3 > G4, suggesting that G9 and G12 had higher Al-tolerance than other two clones. Microscopic observation revealed changes in leaf morphology when exposed to Al stress; for example, a reduced thickness of leaf epidermis and palisade tissue, the descendant palisade tissue/spongy tissue ratio and leaf tissue looseness. Overall, the acid and aluminum stress exerted negative effects on the photosynthetic activity of eucalypt seedlings, but the differences in tolerance to Al toxicity between the clones were favorable, offering potential to improve Eucalyptus plantation productivity by selecting Al tolerant clones. PMID

  20. 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...

  1. Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning.

    Science.gov (United States)

    Tuo, Decai; Shen, Wentao; Yan, Pu; Li, Xiaoying; Zhou, Peng

    2015-12-01

    Papaya leaf distortion mosaic virus (PLDMV) is becoming a threat to papaya and transgenic papaya resistant to the related pathogen, papaya ringspot virus (PRSV). The generation of infectious viral clones is an essential step for reverse-genetics studies of viral gene function and cross-protection. In this study, a sequence- and ligation-independent cloning system, the In-Fusion(®) Cloning Kit (Clontech, Mountain View, CA, USA), was used to construct intron-less or intron-containing full-length cDNA clones of the isolate PLDMV-DF, with the simultaneous scarless assembly of multiple viral and intron fragments into a plasmid vector in a single reaction. The intron-containing full-length cDNA clone of PLDMV-DF was stably propagated in Escherichia coli. In vitro intron-containing transcripts were processed and spliced into biologically active intron-less transcripts following mechanical inoculation and then initiated systemic infections in Carica papaya L. seedlings, which developed similar symptoms to those caused by the wild-type virus. However, no infectivity was detected when the plants were inoculated with RNA transcripts from the intron-less construct because the instability of the viral cDNA clone in bacterial cells caused a non-sense or deletion mutation of the genomic sequence of PLDMV-DF. To our knowledge, this is the first report of the construction of an infectious full-length cDNA clone of PLDMV and the splicing of intron-containing transcripts following mechanical inoculation. In-Fusion cloning shortens the construction time from months to days. Therefore, it is a faster, more flexible, and more efficient method than the traditional multistep restriction enzyme-mediated subcloning procedure.

  2. Rapid Construction of Stable Infectious Full-Length cDNA Clone of Papaya Leaf Distortion Mosaic Virus Using In-Fusion Cloning

    Directory of Open Access Journals (Sweden)

    Decai Tuo

    2015-12-01

    Full Text Available Papaya leaf distortion mosaic virus (PLDMV is becoming a threat to papaya and transgenic papaya resistant to the related pathogen, papaya ringspot virus (PRSV. The generation of infectious viral clones is an essential step for reverse-genetics studies of viral gene function and cross-protection. In this study, a sequence- and ligation-independent cloning system, the In-Fusion® Cloning Kit (Clontech, Mountain View, CA, USA, was used to construct intron-less or intron-containing full-length cDNA clones of the isolate PLDMV-DF, with the simultaneous scarless assembly of multiple viral and intron fragments into a plasmid vector in a single reaction. The intron-containing full-length cDNA clone of PLDMV-DF was stably propagated in Escherichia coli. In vitro intron-containing transcripts were processed and spliced into biologically active intron-less transcripts following mechanical inoculation and then initiated systemic infections in Carica papaya L. seedlings, which developed similar symptoms to those caused by the wild-type virus. However, no infectivity was detected when the plants were inoculated with RNA transcripts from the intron-less construct because the instability of the viral cDNA clone in bacterial cells caused a non-sense or deletion mutation of the genomic sequence of PLDMV-DF. To our knowledge, this is the first report of the construction of an infectious full-length cDNA clone of PLDMV and the splicing of intron-containing transcripts following mechanical inoculation. In-Fusion cloning shortens the construction time from months to days. Therefore, it is a faster, more flexible, and more efficient method than the traditional multistep restriction enzyme-mediated subcloning procedure.

  3. Use of high throughput qPCR screening to rapidly clone low frequency tumour specific T-cells from peripheral blood for adoptive immunotherapy

    Directory of Open Access Journals (Sweden)

    Serrano Oscar K

    2008-10-01

    Full Text Available Abstract Background The adoptive transfer of autologous tumor reactive lymphocytes can mediate significant tumor regression in some patients with refractory metastatic cancer. However, a significant obstacle for this promising therapy has been the availability of highly efficient methods to rapidly isolate and expand a variety of potentially rare tumor reactive lymphocytes from the natural repertoire of cancer patients. Methods We developed a novel in vitro T cell cloning methodology using high throughput quantitative RT-PCR (qPCR assay as a rapid functional screen to detect and facilitate the limiting dilution cloning of a variety of low frequency T cells from bulk PBMC. In preclinical studies, this strategy was applied to the isolation and expansion of gp100 specific CD8+ T cell clones from the peripheral blood of melanoma patients. Results In optimization studies, the qPCR assay could detect the reactivity of 1 antigen specific T cell in 100,000 background cells. When applied to short term sensitized PBMC microcultures, this assay could detect T cell reactivity against a variety of known melanoma tumor epitopes. This screening was combined with early limiting dilution cloning to rapidly isolate gp100154–162 reactive CD8+ T cell clones. These clones were highly avid against peptide pulsed targets and melanoma tumor lines. They had an effector memory phenotype and showed significant proliferative capacity to reach cell numbers appropriate for adoptive transfer trials (~1010 cells. Conclusion This report describes a novel high efficiency strategy to clone tumor reactive T cells from peripheral blood for use in adoptive immunotherapy.

  4. 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.

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

    Science.gov (United States)

    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.

  6. 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.

  7. 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.

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

    Science.gov (United States)

    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 ).

  9. 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.

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

    Science.gov (United States)

    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).

  11. 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

  12. 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).

  13. 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).

  14. 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

  15. Fusion primer and nested integrated PCR (FPNI-PCR): a new high-efficiency strategy for rapid chromosome walking or flanking sequence cloning

    Science.gov (United States)

    2011-01-01

    Background The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created, such as rice and Arabidopsis. The currently available methods for flanking sequence cloning, including the popular TAIL-PCR technique, are relatively laborious and slow. Results Here, we report a simple and effective fusion primer and nested integrated PCR method (FPNI-PCR) for the identification and cloning of unknown genomic regions flanked known sequences. In brief, a set of universal primers was designed that consisted of various 15-16 base arbitrary degenerate oligonucleotides. These arbitrary degenerate primers were fused to the 3' end of an adaptor oligonucleotide which provided a known sequence without degenerate nucleotides, thereby forming the fusion primers (FPs). These fusion primers are employed in the first step of an integrated nested PCR strategy which defines the overall FPNI-PCR protocol. In order to demonstrate the efficacy of this novel strategy, we have successfully used it to isolate multiple genomic sequences namely, 21 orthologs of genes in various species of Rosaceace, 4 MYB genes of Rosa rugosa, 3 promoters of transcription factors of Petunia hybrida, and 4 flanking sequences of T-DNA insertion sites in transgenic tobacco lines and 6 specific genes from sequenced genome of rice and Arabidopsis. Conclusions The successful amplification of target products through FPNI-PCR verified that this novel strategy is an effective, low cost and simple procedure. Furthermore, FPNI-PCR represents a more sensitive, rapid and accurate technique than the established TAIL-PCR and hiTAIL-PCR procedures. PMID:22093809

  16. Rapid containment of nosocomial transmission of a rare community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) clone, responsible for the Staphylococcal Scalded Skin Syndrome (SSSS).

    Science.gov (United States)

    Lamanna, Onofrio; Bongiorno, Dafne; Bertoncello, Lisa; Grandesso, Stefano; Mazzucato, Sandra; Pozzan, Giovanni Battista; Cutrone, Mario; Chirico, Michela; Baesso, Flavia; Brugnaro, Pierluigi; Cafiso, Viviana; Stefani, Stefania; Campanile, Floriana

    2017-01-06

    The aims of this study were to identify the source and the transmission pathway for a Staphylococcal Scalded Skin Syndrome (SSSS) outbreak in a maternity setting in Italy over 2 months, during 2014; to implement appropriate control measures in order to prevent the epidemic spread within the maternity ward; and to identify the Methicillin-Resistant Staphylococcus aureus (MRSA) epidemic clone. Epidemiological and microbiological investigations, based on phenotyping and genotyping methods, were performed. All neonates involved in the outbreak underwent clinical and microbiological investigations to detect the cause of illness. Parents and healthcare workers were screened for Staphylococcus aureus to identify asymptomatic carriers. The SSSS outbreak was due to the cross-transmission of a rare clone of ST5-CA-MRSA-SCCmecV-spa type t311, exfoliative toxin A-producer, isolated from three neonates, one mother (from her nose and from dermatological lesions due to pre-existing hand eczema) and from a nurse (colonized in her nose by this microorganism). The epidemiological and microbiological investigation confirmed these as two potential carriers. A rapid containment of these infections was obtained only after implementation of robust swabbing of mothers and healthcare workers. The use of molecular methodologies for typing was able to identify all carriers and to trace the transmission.

  17. Performance of a PCR assay for the rapid identification of the Klebsiella pneumoniae ST258 epidemic clone in Latin American clinical isolates.

    Science.gov (United States)

    Gomez, S A; Rapoport, M; Piergrossi, N; Faccone, D; Pasteran, F; De Belder, D; ReLAVRA-Group; Petroni, A; Corso, A

    2016-10-01

    The worldwide dissemination of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae ST258 demands a rapid PCR-based typing method to detect unique genes of the ST258 clone. This study evaluates a PCR developed by Adler et al. (2014) for the detection of ST258 in K. pneumoniae clinical isolates centered on the identification of the pilv-I and prp genes. We tested 143 clinical isolates from Argentina (n=109), Chile (n=1), Colombia (n=1), Costa Rica (n=2), Ecuador (n=5), El Salvador (n=2), Nicaragua (n=5), Panamá (n=2), Paraguay (n=2), Perú (n=3) and Trinidad and Tobago (n=11) recovered from 2006 to 2015. blaKPC, pilv-l and prp genes were detected by PCR and sequenced by standard procedures. ST258 and non-ST258 were defined by PFGE and/or MLST. Isolates were grouped according to PFGE patterns: 58 were compatible with ST258 (group 1) and 85 with non-ST258 (group 2). MLST study was done on an arbitrary selection of isolates. The pilv-l gene was present only in ST258 isolates, regardless of the presence of the blaKPC gene. Results for the prp gene were variable. Its presence did not define ST258. The pilv-I PCR had a sensitivity and specificity of 100%, respectively, for the detection of ST258 in the isolates under investigation. Given our findings, the pilv-I PCR could replace more time and resource consuming methods, allowing for more rapid detection of the circulating high risk K. pneumoniae clone ST258 in Latin American (LA) countries. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. 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...

  19. 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.

  20. 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.

  1. Rapid differentiation of Listeria monocytogenes epidemic clones III and IV and their intact compared with heat-killed populations using Fourier transform infrared spectroscopy and chemometrics.

    Science.gov (United States)

    Nyarko, Esmond B; Puzey, Kenneth A; Donnelly, Catherine W

    2014-06-01

    The objectives of this study were to determine if Fourier transform infrared (FT-IR) spectroscopy and multivariate statistical analysis (chemometrics) could be used to rapidly differentiate epidemic clones (ECs) of Listeria monocytogenes, as well as their intact compared with heat-killed populations. FT-IR spectra were collected from dried thin smears on infrared slides prepared from aliquots of 10 μL of each L. monocytogenes ECs (ECIII: J1-101 and R2-499; ECIV: J1-129 and J1-220), and also from intact and heat-killed cell populations of each EC strain using 250 scans at a resolution of 4 cm(-1) in the mid-infrared region in a reflectance mode. Chemometric analysis of spectra involved the application of the multivariate discriminant method for canonical variate analysis (CVA) and linear discriminant analysis (LDA). CVA of the spectra in the wavelength region 4000 to 600 cm(-1) separated the EC strains while LDA resulted in a 100% accurate classification of all spectra in the data set. Further, CVA separated intact and heat-killed cells of each EC strain and there was 100% accuracy in the classification of all spectra when LDA was applied. FT-IR spectral wavenumbers 1650 to 1390 cm(-1) were used to separate heat-killed and intact populations of L. monocytogenes. The FT-IR spectroscopy method allowed discrimination between strains that belong to the same EC. FT-IR is a highly discriminatory and reproducible method that can be used for the rapid subtyping of L. monocytogenes, as well as for the detection of live compared with dead populations of the organism. Fourier transform infrared (FT-IR) spectroscopy and multivariate statistical analysis can be used for L. monocytogenes source tracking and for clinical case isolate comparison during epidemiological investigations since the method is capable of differentiating epidemic clones and it uses a library of well-characterized strains. The FT-IR method is potentially less expensive and more rapid compared to genetic

  2. 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.

  3. What is Cloning?

    Science.gov (United States)

    Donate Home Cloning What is Cloning What is Cloning Clones are organisms that are exact genetic copies. ... clones made through modern cloning technologies. How Is Cloning Done? Many people first heard of cloning when ...

  4. 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.

  5. 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.

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

    Science.gov (United States)

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

    2013-12-01

    concept of our modeling approach and describe its strengths and weaknesses. We furthermore use empirical data for the states of North and South Carolina to demonstrate how the modeling framework can be applied to a large, heterogeneous study system with diverse decision-making agents. Grimm et al. (2005) Pattern-Oriented Modeling of Agent-Based Complex Systems: Lessons from Ecology. Science 310, 987-991. Liu et al. (2013) Framing Sustainability in a Telecoupled World. Ecology and Society 18(2), 26. Meentemeyer et al. (2013) FUTURES: Multilevel Simulations of Merging Urban-Rural Landscape Structure Using a Stochastic Patch-Growing Algorithm. Annals of the Association of American Geographers 103(4), 785-807.

  7. [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...

  8. Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    Science.gov (United States)

    Throop, Andrea L.; LaBaer, Joshua

    2015-01-01

    The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full-length cDNAs in species-specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site-specific recombination that is independent of the insert DNA sequence of interest, which differentiates this method from the classical restriction enzyme-based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression-ready clones using the most popular recombinational cloning technologies, including the commercially available Gateway® (Life Technologies) and In-Fusion® (Clontech) cloning technologies. PMID:25827088

  9. cDNA cloning and chromosomal mapping of the mouse type VII collagen gene (Col7a1): Evidence for rapid evolutionary divergence of the gene

    Energy Technology Data Exchange (ETDEWEB)

    Li, Kehua; Christiano, A.M.; Chu, Mon Li; Uitto, J. (Jefferson Medical College, Philadelphia, PA (United States) Thomas Jefferson Univ., Philadelphia, PA (United States)); Copeland, N.G.; Gilbert, D.J. (NCI-Federick Cancer Research and Development Center, Federick, MD (United States))

    1993-06-01

    Type VII collagen is the major component of anchoring fibrils, critical attachment structures at the dermal-epidermal basement membrane zone. Genetic linkage analyses with recently cloned human type VII collagen cDNAs have indicated that the corresponding gene, COL7A1, is the candidate gene in the dystrophic forms of epidermolysis bullosa. To gain insight into the evolutionary conservation of COL7A1, in this study the authors have isolated mouse type VII collagen cDNAs by screening a mouse epidermal keratinocyte cDNA library with a human COL7A1 cDNA. Two overlapping mouse cDNAs were isolated, and Northern hybridization of mouse epidermal keratinocyte RNA with one of them revealed the presence of a mRNA transcript of [approximately]9.5 kb, the approximate size of the human COL7A1 mRNA. Nucleotide sequencing of the mouse cDNAs revealed a 2760-bp open reading frame that encodes the 5[prime] half of the collagenous domain and a segment of the NC-1, the noncollagenous amino-terminal domain of type VII collagen. Comparison of the mouse amino acid sequences with the corresponding human sequences deduced from cDNAs revealed 82.5% identity. The evolutionary divergence of the gene was relatively rapid in comparison to other collagen genes. Despite the high degree of sequence variation, several sequences, including the size and the position of noncollagenous imperfections and interruptions within the Gly-X-Y repeat sequence, were precisely conserved. Finally, the mouse Col7a1 gene was located by interspecific backcross mapping to mouse Chromosome 9, a region that corresponds to human chromosome 3p21, the position of human COL7Al. This assignment confirms and extends the relationship between the mouse and the human chromosomes in this region of the genome. 33 refs., 5 figs., 1 tab.

  10. Academic Cloning.

    Science.gov (United States)

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

    The authors define "cloning" as an integral feature of all educational systems, citing teaching practices which reward students for closely reproducing the teacher's thoughts and/or behaviors and administrative systems which tend to promote like-minded subordinates. They insist, however, that "academic cloning" is not a totally…

  11. 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.

  12. 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.

  13. Why Clone?

    Science.gov (United States)

    ... to clone agricultural animals, such as cattle and pigs, that are efficient producers of high-quality milk ... risks involved. Visit Teach.Genetics Cite this page Feedback Disclaimer Permissions Policy Privacy Policy Copyright © University of ...

  14. 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.

  15. 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.

  16. Cloning, clones and clonal disease.

    Science.gov (United States)

    Luzzatto, L

    2000-01-01

    In the past, cloning has been familiar to plant breeders because many plants can be easily reproduced in this way, bypassing the lengthy process of cross-fertilisation. Recently, the concept of cloning has become popular in human biology and medicine on two accounts. First, individual genes can be cloned from the enormous complexity of the DNA that makes up the human genetic material. It is expected that, within a few years, all the estimated 100,000 human genes will be isolated by this approach. This should make it possible to identify all the genes that determine the individual characteristics of human beings, including those responsible for causing human diseases or for making people more or less susceptible to pick up diseases from the environment. Cloned genes made into pharmaceutical products are already in use for treating a variety of diseases, from hormonal deficiencies to certain types of anaemia.

  17. Emergence of clones in sexual populations

    Science.gov (United States)

    Neher, Richard A.; Vucelja, Marija; Mezard, Mark; Shraiman, Boris I.

    2013-01-01

    In sexual population, recombination reshuffles genetic variation and produces novel combinations of existing alleles, while selection amplifies the fittest genotypes in the population. If recombination is more rapid than selection, populations consist of a diverse mixture of many genotypes, as is observed in many populations. In the opposite regime, which is realized for example in the facultatively sexual populations that outcross in only a fraction of reproductive cycles, selection can amplify individual genotypes into large clones. Such clones emerge when the fitness advantage of some of the genotypes is large enough that they grow to a significant fraction of the population despite being broken down by recombination. The occurrence of this ‘clonal condensation’ depends, in addition to the outcrossing rate, on the heritability of fitness. Clonal condensation leads to a strong genetic heterogeneity of the population which is not adequately described by traditional population genetics measures, such as linkage disequilibrium. Here we point out the similarity between clonal condensation and the freezing transition in the random energy model of spin glasses. Guided by this analogy we explicitly calculate the probability, Y, that two individuals are genetically identical as a function of the key parameters of the model. While Y is the analog of the spin-glass order parameter, it is also closely related to rate of coalescence in population genetics: two individuals that are part of the same clone have a recent common ancestor.

  18. Human Cloning

    Science.gov (United States)

    2006-07-20

    research group, headed by Douglas Melton and Kevin Eggan, submitted their proposal to a Harvard committee composed of ethicists, scientists and public...United States. Although the company offered no proof of its claim, Dr . Brigette Boisselier, Managing Director of Clonaid, stated that genetic tests would...a year of the Dolly announcement, concerns over human cloning were heightened when Dr . Richard Seed, a Chicago scientist, announced on January 7

  19. 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...

  20. Telomeres and the ethics of human cloning.

    Science.gov (United States)

    Allhoff, Fritz

    2004-01-01

    In search of a potential problem with cloning, I investigate the phenomenon of telomere shortening which is caused by cell replication; clones created from somatic cells will have shortened telomeres and therefore reach a state of senescence more rapidly. While genetic intervention might fix this problem at some point in the future, I ask whether, absent technological advances, this biological phenomenon undermines the moral permissibility of cloning.

  1. 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.

  2. A visual method for direct selection of high-producing Pichia pastoris clones

    Directory of Open Access Journals (Sweden)

    Liu Sheng

    2011-03-01

    Full Text Available Abstract Background The methylotrophic yeast, Pichia pastoris, offers the possibility to generate a high amount of recombinant proteins in a fast and easy way to use expression system. Being a single-celled microorganism, P. pastoris is easy to manipulate and grows rapidly on inexpensive media at high cell densities. A simple and direct method for the selection of high-producing clones can dramatically enhance the whole production process along with significant decrease in production costs. Results A visual method for rapid selection of high-producing clones based on mannanase reporter system was developed. The study explained that it was possible to use mannanase activity as a measure of the expression level of the protein of interest. High-producing target protein clones were directly selected based on the size of hydrolysis holes in the selected plate. As an example, the target gene (9elp-hal18 was expressed and purified in Pichia pastoris using this technology. Conclusions A novel methodology is proposed for obtaining the high-producing clones of proteins of interest, based on the mannanase reporter system. This system may be adapted to other microorganisms, such as Saccharomyces cerevisiae for the selection of clones.

  3. 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.

  4. 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.

  5. Human cloning 2001.

    Science.gov (United States)

    Healy, David L; Weston, Gareth; Pera, Martin F; Rombauts, Luk; Trounson, Alan O

    2002-05-01

    This review summaries human cloning from a clinical perspective. Natural human clones, that is, monozygotic twins, are increasing in the general community. Iatrogenic human clones have been produced for decades in infertile couples given fertility treatment such as ovulation induction. A clear distinction must be made between therapeutic cloning using embryonic stem cells and reproductive cloning attempts. Unlike the early clinical years of in vitro fertilization, with cloning there is no animal model that is safe and dependable. Until there is such a model, 'Dolly'-style human cloning is medically unacceptable.

  6. Cloning of observables

    OpenAIRE

    Ferraro, Alessandro; Galbiati, Matteo; Paris, Matteo G. A.

    2005-01-01

    We introduce the concept of cloning for classes of observables and classify cloning machines for qubit systems according to the number of parameters needed to describe the class under investigation. A no-cloning theorem for observables is derived and the connections between cloning of observables and joint measurements of noncommuting observables are elucidated. Relationships with cloning of states and non-demolition measurements are also analyzed.

  7. 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...

  8. 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.

  9. 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.

  10. Recombination-assisted megaprimer (RAM) cloning

    Science.gov (United States)

    Mathieu, Jacques; Alvarez, Emilia; Alvarez, Pedro J.J.

    2014-01-01

    No molecular cloning technique is considered universally reliable, and many suffer from being too laborious, complex, or expensive. Restriction-free cloning is among the simplest, most rapid, and cost-effective methods, but does not always provide successful results. We modified this method to enhance its success rate through the use of exponential amplification coupled with homologous end-joining. This new method, recombination-assisted megaprimer (RAM) cloning, significantly extends the application of restriction-free cloning, and allows efficient vector construction with much less time and effort when restriction-free cloning fails to provide satisfactory results. The following modifications were made to the protocol:•Limited number of PCR cycles for both megaprimer synthesis and the cloning reaction to reduce error propagation.•Elimination of phosphorylation and ligation steps previously reported for cloning methods that used exponential amplification, through the inclusion of a reverse primer in the cloning reaction with a 20 base pair region of homology to the forward primer.•The inclusion of 1 M betaine to enhance both reaction specificity and yield. PMID:26150930

  11. Cloning of endangered mammalian species: any progress?

    Science.gov (United States)

    Loi, Pasqualino; Galli, Cesare; Ptak, Grazyna

    2007-05-01

    Attempts through somatic cell nuclear transfer to expand wild populations that have shrunk to critical numbers is a logical extension of the successful cloning of mammals. However, although the first mammal was cloned 10 years ago, nuclear reprogramming remains phenomenological, with abnormal gene expression and epigenetic deregulation being associated with the cloning process. In addition, although cloning of wild animals using host oocytes from different species has been successful, little is known about the implication of partial or total mitochondrial DNA heteroplasmy in cloned embryos, fetuses and offspring. Finally, there is a need for suitable foster mothers for inter-intra specific cloned embryos. Considering these issues, the limited success achieved in cloning endangered animals is not surprising. However, optimism comes from the rapid gain in the understanding of the molecular clues underlying nuclear reprogramming. If it is possible to achieve a controlled reversal of the differentiated state of a cell then it is probable that other issues that impair the cloning of endangered animals, such as the inter-intra species oocyte or womb donor, will be overcome in the medium term.

  12. Empirical Results on Cloning and Clone Detection

    OpenAIRE

    Wagner, Stefan

    2015-01-01

    Cloning means the use of copy-paste as method in developing software artefacts. This practice has several problems, such as unnecessary increase of these artefacts, and thereby increased comprehension and change efforts, as well as potential inconsistencies. The automatic detection of clones has been a topic for research for several years now and we have made huge progress in terms of precision and recall. This led to a series of empirical analyses we have performed on the effects and the amo...

  13. Transposon-containing DNA cloning vector and uses thereof

    Science.gov (United States)

    Berg, Claire M.; Berg, Douglas E.; Wang, Gan

    1997-01-01

    The present invention discloses a rapid method of restriction mapping, sequencing or localizing genetic features in a segment of deoxyribonucleic acid (DNA) that is up to 42 kb in size. The method in part comprises cloning of the DNA segment in a specialized cloning vector and then isolating nested deletions in either direction in vivo by intramolecular transposition into the cloned DNA. A plasmid has been prepared and disclosed.

  14. Statement on Human Cloning

    Science.gov (United States)

    ... as our understanding of this technology advances. Support Stem Cell Research (including Research Cloning) AAAS supports stem cell research, including the use of nuclear transplantation techniques (also ...

  15. Gateway Recombinational Cloning.

    Science.gov (United States)

    Reece-Hoyes, John S; Walhout, Albertha J M

    2018-01-02

    The Gateway recombinatorial cloning system was developed for cloning multiple DNA fragments in parallel (e.g., in 96-well formats) in a standardized manner using the same enzymes. Gateway cloning is based on the highly specific integration and excision reactions of bacteriophage λ into and out of the Escherichia coli genome. Because the sites of recombination (" att " sites) are much longer (25-242 bp) than restriction sites, they are extremely unlikely to occur by chance in DNA fragments. Therefore, the same recombination enzyme can be used to robustly clone many different fragments of variable size in parallel reactions. © 2018 Cold Spring Harbor Laboratory Press.

  16. 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

  17. High-throughput cloning and expression in recalcitrant bacteria

    NARCIS (Netherlands)

    Geertsma, Eric R.; Poolman, Bert

    We developed a generic method for high-throughput cloning in bacteria that are less amenable to conventional DNA manipulations. The method involves ligation-independent cloning in an intermediary Escherichia coli vector, which is rapidly converted via vector-backbone exchange (VBEx) into an

  18. 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)].

  19. Microsatellite Primers for Fungus-Growing Ants

    DEFF Research Database (Denmark)

    Villesen Fredsted, Palle; Gertsch, Pia J.; Boomsma, Jacobus Jan (Koos)

    2002-01-01

    We isolated five polymorphic microsatellite loci from a library of two thousand recombinant clones of two fungus-growing ant species, Cyphomyrmex longiscapus and Trachymyrmex cf. zeteki. Amplification and heterozygosity were tested in five species of higher attine ants using both the newly...... developed primers and earlier published primers that were developed for fungus-growing ants. A total of 20 variable microsatellite loci, developed for six different species of fungus-growing ants, are now available for studying the population genetics and colony kin-structure of these ants....

  20. Microsatellite primers for fungus-growing ants

    DEFF Research Database (Denmark)

    Villesen, Palle; Gertsch, P J; Boomsma, JJ

    2002-01-01

    We isolated five polymorphic microsatellite loci from a library of two thousand recombinant clones of two fungus-growing ant species, Cyphomyrmex longiscapus and Trachymyrmex cf. zeteki. Amplification and heterozygosity were tested in five species of higher attine ants using both the newly...... developed primers and earlier published primers that were developed for fungus-growing ants. A total of 20 variable microsatellite loci, developed for six different species of fungus-growing ants, are now available for studying the population genetics and colony kin-structure of these ants....

  1. Cloning-free CRISPR

    NARCIS (Netherlands)

    Arbab, Mandana; Srinivasan, Sharanya; Hashimoto, Tatsunori; Geijsen, Niels; Sherwood, Richard I

    2015-01-01

    We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each

  2. Cloning, killing, and identity.

    Science.gov (United States)

    McMahan, J

    1999-01-01

    One potentially valuable use of cloning is to provide a source of tissues or organs for transplantation. The most important objection to this use of cloning is that a human clone would be the sort of entity that it would be seriously wrong to kill. I argue that entities of the sort that you and I essentially are do not begin to exist until around the seventh month of fetal gestation. Therefore to kill a clone prior to that would not be to kill someone like you or me but would be only to prevent one of us from existing. And even after one of us begins to exist, the objections to killing it remain comparatively weak until its psychological capacities reach a certain level of maturation. These claims support the permissibility of killing a clone during the early stages of its development in order to use its organs for transplantation. PMID:10226909

  3. 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

  4. 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…

  5. Survival and growth of 31 Populus clones in South Carolina

    Energy Technology Data Exchange (ETDEWEB)

    Coyle, David R.; Coleman, Mark D. [USDA Forest Service, Southern Research Station, P.O. Box 700, New Ellenton, SC 29809 (United States); Durant, Jaclin A.; Newman, Lee A. [Arnold School of Public Health, Department of Environmental Health Sciences, University of South Carolina, 800 Sumter Street, Columbia, SC 29208 (United States)

    2006-08-15

    Populus species and hybrids have many practical applications, but clonal performance is relatively undocumented in the southeastern United States outside of the Mississippi River alluvial floodplain. In spring 2001, 31 Populus clones were planted on two sites in South Carolina, USA. The sandy, upland site received irrigation and fertilization throughout the growing season, while the bottomland site received granular fertilizer yearly and irrigation in the first two years only. Over three growing seasons, tree survival and growth differed significantly among clones at both sites. Hybrid clones I45/51, Eridano, and NM6 had very high survival at both sites, while pure eastern cottonwood (P. deltoides) clones consistently had the lowest survival. Nearly all mortality occurred during the first year. The P. deltoides clone WV416 grew well at both sites, P. deltoides clones S13C20 and Kentucky 8 grew well at the bottomland site, and hybrids 184-411 and 52-225 grew well at the upland site. Based on both survival and growth, clones 311-93, S7C15, 184-411, and WV416 may warrant additional testing in the upper coastal plain region of the southeastern US. Kentucky 8 and S13C20 had excellent growth rates, but initial survival was low. However, this was likely due to planting stock quality. We emphasize this is preliminary information, and that clones should be followed through an entire rotation before large-scale deployment. (author)

  6. BIOETHICS AND HUMAN CLONING

    Directory of Open Access Journals (Sweden)

    Željko Kaluđerović

    2011-12-01

    Full Text Available In this paper the authors analyze the process of negotiating and beginning of the United Nations Declaration on Human Cloning as well as the paragraphs of the very Declaration. The negotiation was originally conceived as a clear bioethical debate that should have led to a general agreement to ban human cloning. However, more often it had been discussed about human rights, cultural, civil and religious differences between people and about priorities in case of eventual conflicts between different value systems. In the end, a non-binding Declaration on Human Cloning had been adopted, full of numerous compromises and ambiguous formulations, that relativized the original intention of proposer states. According to authors, it would have been better if bioethical discussion and eventual regulations on cloning mentioned in the following text had been left over to certain professional bodies, and only after the public had been fully informed about it should relevant supranational organizations have taken that into consideration.

  7. Main: Clone Detail [KOME

    Lifescience Database Archive (English)

    Full Text Available Clone Detail Mapping Pseudomolecule data detail Detail information Mapping to the T...IGR japonica Pseudomolecules kome_mapping_pseudomolecule_data_detail.zip kome_mapping_pseudomolecule_data_detail ...

  8. Unified Approach to Universal Cloning and Phase-Covariant Cloning

    OpenAIRE

    Hu, Jia-Zhong; Yu, Zong-Wen; Wang, Xiang-Bin

    2008-01-01

    We analyze the problem of approximate quantum cloning when the quantum state is between two latitudes on the Bloch's sphere. We present an analytical formula for the optimized 1-to-2 cloning. The formula unifies the universal quantum cloning (UQCM) and the phase covariant quantum cloning.

  9. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.|info:eu-repo/dai/nl/074334603; Folkers, G.E.|info:eu-repo/dai/nl/162277202

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning

  10. 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...

  11. Inverse fusion PCR cloning.

    Directory of Open Access Journals (Sweden)

    Markus Spiliotis

    Full Text Available Inverse fusion PCR cloning (IFPC is an easy, PCR based three-step cloning method that allows the seamless and directional insertion of PCR products into virtually all plasmids, this with a free choice of the insertion site. The PCR-derived inserts contain a vector-complementary 5'-end that allows a fusion with the vector by an overlap extension PCR, and the resulting amplified insert-vector fusions are then circularized by ligation prior transformation. A minimal amount of starting material is needed and experimental steps are reduced. Untreated circular plasmid, or alternatively bacteria containing the plasmid, can be used as templates for the insertion, and clean-up of the insert fragment is not urgently required. The whole cloning procedure can be performed within a minimal hands-on time and results in the generation of hundreds to ten-thousands of positive colonies, with a minimal background.

  12. Optimal asymptotic cloning machines

    Science.gov (United States)

    Chiribella, Giulio; Yang, Yuxiang

    2014-06-01

    We pose the question whether the asymptotic equivalence between quantum cloning and quantum state estimation, valid at the single-clone level, still holds when all clones are examined globally. We conjecture that the answer is affirmative and present a large amount of evidence supporting our conjecture, developing techniques to derive optimal asymptotic cloners and proving their equivalence with estimation in virtually all scenarios considered in the literature. Our analysis covers the case of arbitrary finite sets of states, arbitrary families of coherent states, arbitrary phase- and multiphase-covariant sets of states, and two-qubit maximally entangled states. In all these examples we observe that the optimal asymptotic cloners enjoy a universality property, consisting in the fact that scaling of their fidelity does not depend on the specific details of the input states, but only on the number of free parameters needed to specify them.

  13. The Cloning of America.

    Science.gov (United States)

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

    Proposes that the U.S. school system purports to prize human variability, but many educators are engaged in activities that seek to homogenize students. Describes these activities, including diagnosis, labeling, ability grouping, and positive reinforcement. Presents suggestions for counselors to combat sources of cloning and self-validation. (RC)

  14. Applications of quantum cloning

    Science.gov (United States)

    Pomarico, E.; Sanguinetti, B.; Sekatski, P.; Zbinden, H.; Gisin, N.

    2011-10-01

    Quantum Cloning Machines (QCMs) allow for the copying of information, within the limits imposed by quantum mechanics. These devices are particularly interesting in the high-gain regime, i.e., when one input qubit generates a state of many output qubits. In this regime, they allow for the study of certain aspects of the quantum to classical transition. The understanding of these aspects is the root of the two recent applications that we will review in this paper: the first one is the Quantum Cloning Radiometer, a device which is able to produce an absolute measure of spectral radiance. This device exploits the fact that in the quantum regime information can be copied with only finite fidelity, whereas when a state becomes macroscopic, this fidelity gradually increases to 1. Measuring the fidelity of the cloning operation then allows to precisely determine the absolute spectral radiance of the input optical source. We will then discuss whether a Quantum Cloning Machine could be used to produce a state visible by the naked human eye, and the possibility of a Bell Experiment with humans playing the role of detectors.

  15. Self-Cloning CRISPR

    NARCIS (Netherlands)

    Arbab, M.; Sherwood, R. I.

    2016-01-01

    CRISPR/Cas9-gene editing has emerged as a revolutionary technology to easily modify specific genomic loci by designing complementary sgRNA sequences and introducing these into cells along with Cas9. Self-cloning CRISPR/Cas9 (scCRISPR) uses a self-cleaving palindromic sgRNA plasmid (sgPal) that

  16. Secure the Clones

    Science.gov (United States)

    Jensen, Thomas; Kirchner, Florent; Pichardie, David

    Exchanging mutable data objects with untrusted code is a delicate matter because of the risk of creating a data space that is accessible by an attacker. Consequently, secure programming guidelines for Java stress the importance of using defensive copying before accepting or handing out references to an internal mutable object. However, implementation of a copy method (like clone()) is entirely left to the programmer. It may not provide a sufficiently deep copy of an object and is subject to overriding by a malicious sub-class. Currently no language-based mechanism supports secure object cloning. This paper proposes a type-based annotation system for defining modular copy policies for class-based object-oriented programs. A copy policy specifies the maximally allowed sharing between an object and its clone. We present a static enforcement mechanism that will guarantee that all classes fulfill their copy policy, even in the presence of overriding of copy methods, and establish the semantic correctness of the overall approach in Coq. The mechanism has been implemented and experimentally evaluated on clone methods from several Java libraries.

  17. A Seminar on Human Cloning: Cloning in Reproductive Medicine

    OpenAIRE

    Illmensee, Karl

    2001-01-01

    This review article summarizes the historical development of mammalian cloning, presents current advances and presumed risk factors in the field of reproductive cloning, discusses possible clinical applications of therapeutic and diagnostic cloning and outlines prospective commercial trends in pharmacytical cloning. Predictable progress in biotechnology and stem cell engineering should prove to be advantageous for patients' health and for novel benefits in reproductive and regenerative medicine.

  18. 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.

  19. The First Human Cloned Embryo.

    Science.gov (United States)

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  20. [Nuclear transfer and therapeutic cloning].

    Science.gov (United States)

    Xu, Xiao-Ming; Lei, An-Min; Hua, Jin-Lian; Dou, Zhong-Ying

    2005-03-01

    Nuclear transfer and therapeutic cloning have widespread and attractive prospects in animal agriculture and biomedical applications. We reviewed that the quality of oocytes and nuclear reprogramming of somatic donor cells were the main reasons of the common abnormalities in cloned animals and the low efficiency of cloning and showed the problems and outlets in therapeutic cloning, such as some basic problems in nuclear transfer affected clinical applications of therapeutic cloning. Study on isolation and culture of nuclear transfer embryonic stem (ntES) cells and specific differentiation of ntES cells into important functional cells should be emphasized and could enhance the efficiency. Adult stem cells could help to cure some great diseases, but could not replace therapeutic cloning. Ethics also impeded the development of therapeutic cloning. It is necessary to improve many techniques and reinforce the research of some basic theories, then somatic nuclear transfer and therapeutic cloning may apply to agriculture reproduction and benefit to human life better.

  1. Animal Cloning and Food Safety

    Science.gov (United States)

    ... Home For Consumers Consumer Updates Animal Cloning and Food Safety Share Tweet Linkedin Pin it More sharing options ... on clones of livestock animals to evaluate the safety of food from these animals. The resulting report, called a ...

  2. HUMAN CLONING AND FDA REGULATION

    OpenAIRE

    Vosk, Ted

    1998-01-01

    In the February 27, 1997 issue of the journal Nature scientists from Scotland's Roslin Institute reported their successful efforts to clone an adult sheep using differentiated somatic cells from the animal. The clone, named Dolly, was the first instance of the successful cloning of an adult mammal. The shock waves created by the possible ramifications of this development were immediately felt around the world. For the first time, the cloning of an adult human being was no longer being conside...

  3. Three concepts of cloning in human beings.

    Science.gov (United States)

    Cui, Ke-Hui

    2005-07-01

    Human cloning, organ cloning and tissue cloning are various types of cloning that occur at different levels with different methodologies. According to three standards of terminology for an embryo (fertilization through germ cells, development in the uterus and having the potential to produce a human life), tissue cloning and type I organ cloning will not produce an embryo. In contrast, human cloning and type II organ cloning will produce an embryo. Thus, only non-germinal tissue cloning and type I organ cloning are beyond the ethical question and will not change human beings as a species. Using cloned tissues to make new tissues or organs is promising for the future of medicine.

  4. Optimally cloned binary coherent states

    Science.gov (United States)

    Müller, C. R.; Leuchs, G.; Marquardt, Ch.; Andersen, U. L.

    2017-10-01

    Binary coherent state alphabets can be represented in a two-dimensional Hilbert space. We capitalize this formal connection between the otherwise distinct domains of qubits and continuous variable states to map binary phase-shift keyed coherent states onto the Bloch sphere and to derive their quantum-optimal clones. We analyze the Wigner function and the cumulants of the clones, and we conclude that optimal cloning of binary coherent states requires a nonlinearity above second order. We propose several practical and near-optimal cloning schemes and compare their cloning fidelity to the optimal cloner.

  5. Cloning-free CRISPR

    Directory of Open Access Journals (Sweden)

    Mandana Arbab

    2015-11-01

    Full Text Available We present self-cloning CRISPR/Cas9 (scCRISPR, a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA or knockin homology construct for each target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis.

  6. A bacteriophage T4 in vitro system to clone long DNA molecules. Final report, June 1, 1990--January 31, 1996

    Energy Technology Data Exchange (ETDEWEB)

    Rao, V.B.

    1997-09-01

    A summary is presented of the following objectives: development of a bacteriophage T4 in vitro system, and techniques to clone long segments of foreign DNA; development of a giant prohead DNA packaging system that could potentially be used to clone even a megabase size DNA; and development of techniques to rapidly map the cloned DNA inserts.

  7. Flow cytometry-assisted cloning of specific sequence motifs from complex 16S rRNA gene libraries

    DEFF Research Database (Denmark)

    Nielsen, J. L.; Schramm, A.; Engh, G. van den

    2004-01-01

    A How cytometry method was developed for rapid screening and recovery of cloned DNA containing common sequence motifs. This approach, termed fluorescence-activated cell sorting-assisted cloning, was used to recover sequences affiliated with a unique lineage within the Bacteroidetes not abundant i...... in a clone library of environmental 16S rRNA genes....

  8. Ethical issues in animal cloning.

    Science.gov (United States)

    Fiester, Autumn

    2005-01-01

    The issue of human reproductive cloning has recently received a great deal attention in public discourse. Bioethicists, policy makers, and the media have been quick to identify the key ethical issues involved in human reproductive cloning and to argue, almost unanimously, for an international ban on such attempts. Meanwhile, scientists have proceeded with extensive research agendas in the cloning of animals. Despite this research, there has been little public discussion of the ethical issues raised by animal cloning projects. Polling data show that the public is decidedly against the cloning of animals. To understand the public's reaction and fill the void of reasoned debate about the issue, we need to review the possible objections to animal cloning and assess the merits of the anti-animal cloning stance. Some objections to animal cloning (e.g., the impact of cloning on the population of unwanted animals) can be easily addressed, while others (e.g., the health of cloned animals) require more serious attention by the public and policy makers.

  9. Growing in darkness: The etiolated lupin hypocotyls

    OpenAIRE

    Sánchez-Bravo, José; Oliveros-Valenzuela, M Rocío; Nicolás, Carlos; Acosta, Manuel

    2008-01-01

    Epigeal germination of a dicot, like lupin (Lupinus albus L.), produces a seedling with a characteristic hypocotyl, which grows in darkness showing a steep growth gradient with an elongation zone just below the apex. The role of phytohormones, such as auxin and ethylene, in etiolated hypocotyl growth has been the object of our research for some time. The recent cloning and expression of three genes of influx and efflux carriers for polar auxin transport (LaAUX1, LaPIN1 and LaPIN3) reinforces ...

  10. Lessons learned from cloning dogs.

    Science.gov (United States)

    Kim, M J; Oh, H J; Kim, G A; Park, J E; Park, E J; Jang, G; Ra, J C; Kang, S K; Lee, B C

    2012-08-01

    The aim of this article is to review dog cloning research and to suggest its applications based on a discussion about the normality of cloned dogs. Somatic cell nuclear transfer was successfully used for production of viable cloned puppies despite limited understanding of in vitro dog embryo production. Cloned dogs have similar growth characteristics to those born from natural fertilization, with no evidence of serious adverse effects. The offspring of cloned dogs also have similar growth performance and health to those of naturally bred puppies. Therefore, cloning in domestic dogs can be applied as an assisted reproductive technique to conserve endangered species, to treat sterile canids or aged dogs, to improve reproductive performance of valuable individuals and to generate disease model animals. © 2012 Blackwell Verlag GmbH.

  11. 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...

  12. Human cloning and child welfare.

    Science.gov (United States)

    Burley, J; Harris, J

    1999-01-01

    In this paper we discuss an objection to human cloning which appeals to the welfare of the child. This objection varies according to the sort of harm it is expected the clone will suffer. The three formulations of it that we will consider are: 1. Clones will be harmed by the fearful or prejudicial attitudes people may have about or towards them (H1); 2. Clones will be harmed by the demands and expectations of parents or genotype donors (H2); 3. Clones will be harmed by their own awareness of their origins, for example the knowledge that the genetic donor is a stranger (H3). We will show why these three versions of the child welfare objection do not necessarily supply compelling reasons to ban human reproductive cloning. The claim that we will develop and defend in the course of our discussion is that even if it is the case that a cloned child will suffer harms of the type H1-H3, it is none the less permissible to conceive by cloning so long as these cloning-induced welfare deficits are not such as to blight the existence of the resultant child, whoever this may be. PMID:10226914

  13. Evaluation of a pooled strategy for high-throughput sequencing of cosmid clones from metagenomic libraries.

    Directory of Open Access Journals (Sweden)

    Kathy N Lam

    Full Text Available High-throughput sequencing methods have been instrumental in the growing field of metagenomics, with technological improvements enabling greater throughput at decreased costs. Nonetheless, the economy of high-throughput sequencing cannot be fully leveraged in the subdiscipline of functional metagenomics. In this area of research, environmental DNA is typically cloned to generate large-insert libraries from which individual clones are isolated, based on specific activities of interest. Sequence data are required for complete characterization of such clones, but the sequencing of a large set of clones requires individual barcode-based sample preparation; this can become costly, as the cost of clone barcoding scales linearly with the number of clones processed, and thus sequencing a large number of metagenomic clones often remains cost-prohibitive. We investigated a hybrid Sanger/Illumina pooled sequencing strategy that omits barcoding altogether, and we evaluated this strategy by comparing the pooled sequencing results to reference sequence data obtained from traditional barcode-based sequencing of the same set of clones. Using identity and coverage metrics in our evaluation, we show that pooled sequencing can generate high-quality sequence data, without producing problematic chimeras. Though caveats of a pooled strategy exist and further optimization of the method is required to improve recovery of complete clone sequences and to avoid circumstances that generate unrecoverable clone sequences, our results demonstrate that pooled sequencing represents an effective and low-cost alternative for sequencing large sets of metagenomic clones.

  14. Molecular cloning of growth hormone encoding cDNA of Indian ...

    Indian Academy of Sciences (India)

    A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in ...

  15. Cloning the mammoth: a complicated task or just a dream?

    Science.gov (United States)

    Loi, Pasqualino; Saragusty, Joseph; Ptak, Grazyna

    2014-01-01

    Recently there has been growing interest in applying the most advanced embryological tools, particularly cloning, to bring extinct species back to life, with a particular focus on the woolly mammoth (Mammuthus primigenius). Mammoth's bodies found in the permafrost are relatively well preserved, with identifiable nuclei in their tissues. The purpose of this chapter is to review the literature published on the topic, and to present the strategies potentially suitable for a mammoth cloning project, with a frank assessment of their feasibility and the ethical issues involved.

  16. [The discrete horror of cloning].

    Science.gov (United States)

    Guibourg, Ricardo A

    2009-01-01

    The author raises the topic of cloning after the decision of the Argentine government, which concerned for the "dignity of the human person", passed a decree of need and urgency, No. 200/97 (Annex), prohibiting cloning experiments with human beings. Therefore, considering that the topic is so terribly urgent and necessary, the author feels it is timely to consider it.

  17. Molecular cloning, characterization and expression analysis of heat ...

    African Journals Online (AJOL)

    Heat shock protein 90 (HSP90) is a highly conserved protein and plays an important role in maintaining the structure of protein, participating in the immunity and regulating the cell cycle. Using the rapid amplification of cDNA ends (RACE) techniques, the cDNA sequence of HSP90 gene (designated Sp- HSP90) was cloned ...

  18. [Scientific ethics of human cloning].

    Science.gov (United States)

    Valenzuela, Carlos Y

    2005-01-01

    True cloning is fission, budding or other types of asexual reproduction. In humans it occurs in monozygote twinning. This type of cloning is ethically and religiously good. Human cloning can be performed by twinning (TWClo) or nuclear transfer (NTClo). Both methods need a zygote or a nuclear transferred cell, obtained in vitro (IVTec). They are under the IVTec ethics. IVTecs use humans (zygotes, embryos) as drugs or things; increase the risk of malformations; increase development and size of abnormalities and may cause long-term changes. Cloning for preserving extinct (or almost extinct) animals or humans when sexual reproduction is not possible is ethically valid. The previous selection of a phenotype in human cloning violates some ethical principles. NTClo for reproductive or therapeutic purposes is dangerous since it increases the risk for nucleotide or chromosome mutations, de-programming or re-programming errors, aging or malignancy of the embryo cells thus obtained.

  19. Field performance and biomass production of 12 willow and poplar clones in short-rotation coppice in southern Quebec (Canada)

    Energy Technology Data Exchange (ETDEWEB)

    Labrecque, Michel; Teodorescu, Traian I. [Montreal Botanical Garden, Inst. de Recherche en Biologie Vegetale, Montreal, QC (Canada)

    2005-07-01

    Twelve clones of fast growing trees (willow and poplar) were planted in 1999 under short-rotation intensive culture (SRIC) on an abandoned farmland in southern Quebec. The plantation was established at a density of 18,000 trees per hectare from stem cuttings and no fertilizer and irrigation were applied. Tree performances were measured at regular intervals during four growing seasons. The aims of the experiment were to compare the growth, insect and disease resistance of these clones in order to select those that have good potential for use as commercial biomass energy crops in northern regions of North America. The follow up of the growing performance has shown statistically significant differences between the clones. Poplar clones registered the highest above ground biomass yield after 4 growing seasons (from 66.48 to 72.20 tDM ha{sup -1}). The best willow biomass productivity was obtained from clones SX64 (67.58 tDM ha{sup -1}) and clone SX61 (62.34 tDM ha{sup -1}). Only one willow clone S301 (Salix interior x S. eriocephala) was sensitive to leaf rust (Melampsora spp.) and clones SVQ (S. viminalis) and SV1 (S. dasyclados) were more prone to insect attacks. The results proved that some clones of S. miyabeana and S. sachalinensis were more productive and more resistant to insect and disease damage than S. viminalis which has been widely planted in SRIC in southern Quebec for many years. (Author)

  20. Animal cloning: problems and prospects.

    Science.gov (United States)

    Wells, D N

    2005-04-01

    An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred to the reproductive tracts of recipient cows result in healthy, longterm surviving clones. Of concern are the high losses throughout gestation, during birth and in the post-natal period through to adulthood. Many of the pregnancy losses relate to failure of the placenta to develop and function correctly. Placental dysfunction may also have an adverse influence on postnatal health. These anomalies are probably due to incorrect epigenetic reprogramming of the donor genome following NT, leading to inappropriate patterns of gene expression during the development of clones. Whilst some physiological tests on surviving clones suggest normality, other reports indicate a variety of post-natal clone-associated abnormalities. This variability in outcome may reflect species-specific and/or cloning methodological differences. Importantly, to date it appears that these clone-associated phenotypes are not transmitted to offspring following sexual reproduction. This indicates that they represent epigenetic errors, rather than genetic errors, which are corrected during gametogenesis. Whilst this needs confirmation at the molecular level, it provides initial confidence in the first application of NT in agriculture, namely, the production of small numbers of cloned sires from genetically elite bulls, for natural mating, to effectively disseminate genetic gain. In addition to the animal welfare concerns with the technology, the underlying health of the animals and the consequential effect on food safety are critical aspects that require investigation to gain regulatory and consumer

  1. [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.

  2. 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).…

  3. 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)

  4. 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…

  5. 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...

  6. HPLC/MS-TOF Analysis of Surface Resins from Three Poplar Clones Grown in Serbia

    Directory of Open Access Journals (Sweden)

    Branislav Trudić

    2016-12-01

    Full Text Available Background and Purpose: Poplar clones grown in Serbia are fast growing tree species important for many different purposes in forestry and industry. In this study chemical content of the surface resins of three poplar clones grown in Serbia - M1, B229 and PE 19/66 was analyzed, aiming at their potential usage as a source of natural products important for pharmacy and chemotaxonomy. Materials and Methods: Using HPLC/MS-TOF we gained the first information on chemical compounds which comprise of resins on terminal twigs cuttings of commonly grown poplar clones. Provided from the nursery of the Institute of Lowland Forestry and Environment (Serbia, terminal twigs cuttings with leaves of different development stage from two year old seedlings of M1 poplar clone (Populus euramericana L., PE 19/66 clone and B229 clone (both belonging to Populus deltoides were sampled. The washing of the surface resins from terminal twigs cuttings of every sample was done with methylene-chloride until the samples were prepared for HPLC/MS-TOF analysis. Results: Out of 38 different compounds which were identified, M1 clone qualitatively differed for 14 compounds as compared to two other clones. Generally, the results showed that the composition of the resins consisted of different phenolic acids, phenolic esters, flavonoids and other contents. Conclusion: These three poplar clones are potent producers of pharmacologically and chemotaxonomically potent compounds in forest ecosystems, especially M1 clone.

  7. Human cloning: can it be made safe?

    Science.gov (United States)

    Rhind, Susan M; Taylor, Jane E; De Sousa, Paul A; King, Tim J; McGarry, Michelle; Wilmut, Ian

    2003-11-01

    There are continued claims of attempts to clone humans using nuclear transfer, despite the serious problems that have been encountered in cloning other mammals. It is known that epigenetic and genetic mechanisms are involved in clone failure, but we still do not know exactly how. Human reproductive cloning is unethical, but the production of cells from cloned embryos could offer many potential benefits. So, can human cloning be made safe?

  8. Quantum Cloning Machines and the Applications

    OpenAIRE

    Fan, Heng; Wang, Yi-Nan; Jing, Li; Yue, Jie-Dong; Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu

    2013-01-01

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the securit...

  9. Post-death cloning of endangered Jeju black cattle (Korean native cattle): fertility and serum chemistry in a cloned bull and cow and their offspring.

    Science.gov (United States)

    Kim, Eun Young; Song, Dong Hwan; Park, Min Jee; Park, Hyo Young; Lee, Seung Eun; Choi, Hyun Yong; Moon, Jeremiah Jiman; Kim, Young Hoon; Mun, Seong Ho; Oh, Chang Eon; Ko, Moon Suck; Lee, Dong Sun; Riu, Key Zung; Park, Se Pill

    2013-12-17

    To preserve Jeju black cattle (JBC; endangered native Korean cattle), a pair of cattle, namely a post-death cloned JBC bull and cow, were produced by somatic cell nuclear transfer (SCNT) in a previous study. In the present study, we examined the in vitro fertilization and reproductive potentials of these post-death cloned animals. Sperm motility, in vitro fertilization and developmental capacity were examined in a post-death cloned bull (Heuk Oll Dolee) and an extinct nuclear donor bull (BK94-13). We assessed reproductive ability in another post-death cloned cow (Heuk Woo Sunee) using cloned sperm for artificial insemination (AI). There were no differences in sperm motility or developmental potential of in vitro fertilized embryos between the post-death cloned bull and its extinct nuclear donor bull; however, the embryo development ratio was slightly higher in the cloned sperm group than in the nuclear donor sperm group. After one attempt at AI, the post-death cloned JBC cow became pregnant, and gestation proceeded normally until day 287. From this post-death cloned sire and dam, a JBC male calf (Heuk Woo Dolee) was delivered naturally (weight, 25 kg). The genetic paternity/maternity of the cloned JBC bull and cow with regard to their offspring was confirmed using International Society for Animal Genetics standard microsatellite markers. Presently, Heuk Woo Dolee is 5 months of age and growing normally. In addition, there were no significant differences in blood chemistry among the post-death cloned JBC bull, the cow, their offspring and cattle bred by AI. This is the first report showing that a pair of cattle, namely, a post-death cloned JBC bull and cow, had normal fertility. Therefore, SCNT can be used effectively to increase the population of endangered JBC.

  10. Limitations on Cloning in Classical Mechanics

    OpenAIRE

    Fenyes, Aaron

    2010-01-01

    In this paper, we show that a result precisely analogous to the traditional quantum no-cloning theorem holds in classical mechanics. This classical no-cloning theorem does not prohibit classical cloning, we argue, because it is based on a too-restrictive definition of cloning. Using a less popular, more inclusive definition of cloning, we give examples of classical cloning processes. We also prove that a cloning machine must be at least as complicated as the object it is supposed to clone.

  11. Therapeutic cloning in the mouse

    Science.gov (United States)

    Mombaerts, Peter

    2003-01-01

    Nuclear transfer technology can be applied to produce autologous differentiated cells for therapeutic purposes, a concept termed therapeutic cloning. Countless articles have been published on the ethics and politics of human therapeutic cloning, reflecting the high expectations from this new opportunity for rejuvenation of the aging or diseased body. Yet the research literature on therapeutic cloning, strictly speaking, is comprised of only four articles, all in the mouse. The efficiency of derivation of embryonic stem cell lines via nuclear transfer is remarkably consistent among these reports. However, the efficiency is so low that, in its present form, the concept is unlikely to become widespread in clinical practice. PMID:12949262

  12. Human therapeutic cloning (NTSC): applying research from mammalian reproductive cloning.

    Science.gov (United States)

    French, Andrew J; Wood, Samuel H; Trounson, Alan O

    2006-01-01

    Human therapeutic cloning or nuclear transfer stem cells (NTSC) to produce patient-specific stem cells, holds considerable promise in the field of regenerative medicine. The recent withdrawal of the only scientific publications claiming the successful generation of NTSC lines afford an opportunity to review the available research in mammalian reproductive somatic cell nuclear transfer (SCNT) with the goal of progressing human NTSC. The process of SCNT is prone to epigenetic abnormalities that contribute to very low success rates. Although there are high mortality rates in some species of cloned animals, most surviving clones have been shown to have normal phenotypic and physiological characteristics and to produce healthy offspring. This technology has been applied to an increasing number of mammals for utility in research, agriculture, conservation, and biomedicine. In contrast, attempts at SCNT to produce human embryonic stem cells (hESCs) have been disappointing. Only one group has published reliable evidence of success in deriving a cloned human blastocyst, using an undifferentiated hESC donor cell, and it failed to develop into a hESC line. When optimal conditions are present, it appears that in vitro development of cloned and parthenogenetic embryos, both of which may be utilized to produce hESCs, may be similar to in vitro fertilized embryos. The derivation of ESC lines from cloned embryos is substantially more efficient than the production of viable offspring. This review summarizes developments in mammalian reproductive cloning, cell-to-cell fusion alternatives, and strategies for oocyte procurement that may provide important clues facilitating progress in human therapeutic cloning leading to the successful application of cell-based therapies utilizing autologous hESC lines.

  13. Rapid cloning and bioinformatic analysis of spinach Y chromosome ...

    Indian Academy of Sciences (India)

    growth environment was maintained to avoid the biotic or abiotic stress. Japanese big leaf spinach with genetic stability ..... inant suppressors of Srs2 helicase mutations of Saccharomyces cerevisiae map in the RAD51 gene, whose sequence predicts a protein with similarities to procaryotic RecA proteins. Mol. Cell. Biol.

  14. Birth of clones of the world's first cloned dog.

    Science.gov (United States)

    Kim, Min Jung; Oh, Hyun Ju; Kim, Geon A; Setyawan, Erif Maha Nugraha; Choi, Yoo Bin; Lee, Seok Hee; Petersen-Jones, Simon M; Ko, CheMyong J; Lee, Byeong Chun

    2017-11-10

    Animal cloning has gained popularity as a method to produce genetically identical animals or superior animals for research or industrial uses. However, the long-standing question of whether a cloned animal undergoes an accelerated aging process is yet to be answered. As a step towards answering this question, we compared longevity and health of Snuppy, the world's first cloned dog, and its somatic cell donor, Tai, a male Afghan hound. Briefly, both Snuppy and Tai were generally healthy until both developed cancer to which they succumbed at the ages of 10 and 12 years, respectively. The longevity of both the donor and the cloned dog was close to the median lifespan of Afghan hounds which is reported to be 11.9 years. Here, we report creation of 4 clones using adipose-derived mesenchymal stem cells from Snuppy as donor cells. Clinical and molecular follow-up of these reclones over their lives will provide us with a unique opportunity to study the health and longevity of cloned animals compared with their cell donors.

  15. Commercial aspects of cloning and genetic modification in cattle

    DEFF Research Database (Denmark)

    Lewis, I M; French, A J; Tecirlioglu, R T

    2004-01-01

    A range of potential commercial applications of cloning and genetic modification in cattle has been suggested over the last decade. It includes the rapid multiplication of elite genotypes, production of valuable human proteins, altered production characteristics, increased disease resistance...... and milk with improved nutritional value and processing capabilities. However, an economic return from the sale of product is far from reality in any of these areas. One impediment to achieving economic sustainability is the extremely low efficiency in producing healthy offspring from transferred cloned...

  16. 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…

  17. Use of belowground growing degree days to predict rooting of dormant hardwood cuttings of Populus

    Science.gov (United States)

    R.S., Jr. Zalesny; E.O. Bauer; D.E. Riemenschneider

    2004-01-01

    Planting Populus cuttings based on calendar days neglects soil temperature extremes and does not promote rooting based on specific genotypes. Our objectives were to: 1) test the biological efficacy of a thermal index based on belowground growing degree days (GDD) across the growing period, 2) test for interactions between belowground GDD and clones,...

  18. Exploring the metal phytoremediation potential of three Populus alba L. clones using an in vitro screening.

    Science.gov (United States)

    Di Lonardo, Sara; Capuana, Maurizio; Arnetoli, Miluscia; Gabbrielli, Roberto; Gonnelli, Cristina

    2011-01-01

    This work was planned for providing a useful screening tool for the selection of Populus alba clones suitable for phytoremediation techniques. To this aim, we investigated variation in arsenic, cadmium, copper, and zinc tolerance, accumulation and translocation in three poplar clones through an in vitro screening. Poplars have been widely proposed for phytoremediation, as they are adaptable to grow on contaminated areas and able to accumulate metals. The investigation of possible differences among poplar clones in metal tolerance and accumulation deserves to be deeply studied and exploited for the selection of the more suitable tool for phytoremediation purposes. In vitro multiplied microshoots of a commercial and two autochthonous P. alba clones were subcultured on hormone-free WPM medium for 1 month and then transferred for 2 weeks onto media containing different concentrations of the metals investigated. At the end of the treatments, plantlets were sampled, weighed, and mineralised by wet ashing. Metal concentrations were determined by ICP-OES. For the metal concentration used in the experiments, our clones of P. alba showed variation in metal tolerance, metal accumulation and content. The fast-growing commercial clone, even if rarely showing the highest plant metal concentration, displayed the highest metal content, suggesting biomass production as the key factor in evaluating the phytoextraction capacity of P. alba clones for the metals studied. Data demonstrated that in vitro screening of cuttings represents a valuable way of assessing the ability of different poplar clones to take up, tolerate and survive metal stress.

  19. 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 ...

  20. 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 ...

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. Comprehensive Cloning of Patient-derived 9022-bp Amplicons of Hepatitis C Virus

    Science.gov (United States)

    Lu, Yang; Xu, Yanjuan; Di Bisceglie, Adrian M.; Fan, Xiaofeng

    2013-01-01

    The instability of recombinant clones accommodating large or full-length viral genomes is frequently a technical challenge in RNA virus research. In an attempt to establish a rapid plasmid-based reverse genetics system that utilizes long RT-PCR technique (LRP), similar difficulty was encountered in the cloning of 9022-bp LRP amplicon. All HCV genotype 1a strains used for LRP cloning showed a remarkable difference in terms of cloning stability. Subsequent analysis revealed the predictive value of phylogenetic positions in determining the cloning stability. Putative E. coli promoters on the HCV genome might be responsible for such cloning difference. An exhaustive exploration, testing nearly one hundred cloning protocols, did not reveal a general approach that can achieve stable cloning for all HCV 1a strains. The selection of appropriate strains, guided by phylogenetic analysis, appears to be necessary prior to the construction of infectious HCV 1a clones. These observations are not only valuable for potentially establishing an HCV 1a cell culture model but also have general implications for other RNA viruses due to concern about cloning instability. PMID:23602804

  4. Human cloning and 'posthuman' society.

    Science.gov (United States)

    Blackford, Russell

    2005-01-01

    Since early 1997, when the creation of Dolly the sheep by somatic cell nuclear transfer was announced in Nature, numerous government reports, essays, articles and books have considered the ethical problems and policy issues surrounding human reproductive cloning. In this article, I consider what response a modern liberal society should give to the prospect of human cloning, if it became safe and practical. Some opponents of human cloning have argued that permitting it would place us on a slippery slope to a repugnant future society, comparable to that portrayed in Aldous Huxley's novel, Brave New World. I conclude that, leaving aside concerns about safety, none of the psychological or social considerations discussed in this article provides an adequate policy justification for invoking the state's coercive powers to prevent human cloning.

  5. Human Cloning: Let's Discuss It.

    Science.gov (United States)

    Taras, Loretta; Stavroulakis, Anthea M.; Ortiz, Mary T.

    1999-01-01

    Describes experiences with holding discussions on cloning at a variety of levels in undergraduate biology courses. Discusses teaching methods used and student reactions to the discussions. Contains 12 references. (WRM)

  6. A novel nucleo-cytoplasmic hybrid clone formed via androgenesis in polyploid gibel carp

    Directory of Open Access Journals (Sweden)

    Zhou Li

    2011-03-01

    Full Text Available Abstract Background Unisexual vertebrates have been demonstrated to reproduce by gynogenesis, hybridogenesis, parthenogenesis, or kleptogenesis, however, it is uncertain how the reproduction mode contributes to the clonal diversity. Recently, polyploid gibel carp has been revealed to possess coexisting dual modes of unisexual gynogenesis and sexual reproduction and to have numerous various clones. Using sexual reproduction mating between clone D female and clone A male and subsequent 7 generation multiplying of unisexual gynogenesis, we have created a novel clone strain with more than several hundred millions of individuals. Here, we attempt to identify genetic background of the novel clone and to explore the significant implication for clonal diversity contribution. Methods Several nuclear genome markers and one cytoplasmic marker, the mitochondrial genome sequence, were used to identify the genetic organization of the randomly sampled individuals from different generations of the novel clone. Results Chromosome number, Cot-1 repetitive DNA banded karyotype, microsatellite patterns, AFLP profiles and transferrin alleles uniformly indicated that nuclear genome of the novel clone is identical to that of clone A, and significantly different from that of clone D. However, the cytoplasmic marker, its complete mtDNA genome sequence, is same to that of clone D, and different from that of clone A. Conclusions The present data indicate that the novel clone is a nucleo-cytoplasmic hybrid between the known clones A and D, because it originates from the offspring of gonochoristic sexual reproduction mating between clone D female and clone A male, and contains an entire nuclear genome from the paternal clone A and a mtDNA genome (cytoplasm from the maternal clone D. It is suggested to arise via androgenesis by a mechanism of ploidy doubling of clone A sperm in clone D ooplasm through inhibiting the first mitotic division. Significantly, the selected nucleo

  7. Islamic perspectives on human cloning.

    Science.gov (United States)

    Sadeghi, Mahmoud

    2007-01-01

    The present paper seeks to assess various views from Islamic jurists relating to human cloning, which is one of the controversial topics in the recent past. Taking Islamic jurisprudence principles, such as the rule of necessity for self preservation and respect for human beings, the rule of la darar wa la dirar ('the necessity to refrain from causing harm to oneself and others') and the rule of usr wa haraj, one may indicate that if human cloning could not be prohibited, as such, it could still be opposed because it gives way to various harmful consequences, which include family disorder, chaos in the clone's family relationships, physical and mental diseases for clones and suffering of egg donors and surrogate mothers. However with due attention to the fact that the reasons behind the prohibition of abortion only restrict the destruction of human embryos in their post-implantation stages, human cloning for biomedical research and exploitation of stem cells from cloned embryos at the blastocyst stage for therapeutic purposes would be acceptable.

  8. Cloning goes to the movies.

    Science.gov (United States)

    Cormick, Craig

    2006-10-01

    Public attitude research conducted by Biotechnology Australia shows that one of the major sources of information on human reproductive cloning is movies. Traditionally, understanding of new and emerging technologies has come through the mass media but human cloning, being so widely addressed through the popular culture of movies, is more effectively defined by Hollywood than the news media or science media. But how well are the science and social issues of cloning portrayed in box office hits such as The Island, Multiplicity, Star Wars: Attack of the Clones and Jurassic Park? These movies have enormous reach and undoubted influence, and are therefore worth analyzing in some detail. This study looks at 33 movies made between 1971 and 2005 that address human reproductive cloning, and it categorizes the films based on their genre and potential influence. Yet rather than simply rating the quality of the science portrayed, the study compares the key messages in these movies with public attitudes towards cloning, to examine the correlations.

  9. Artificial cloning of domestic animals.

    Science.gov (United States)

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research.

  10. Human cloning, stem cell research. An Islamic perspective.

    Science.gov (United States)

    Al-Aqeel, Aida I

    2009-12-01

    The rapidly changing technologies that involve human subjects raise complex ethical, legal, social, and religious issues. Recent advances in the field of cloning and stem cell research have introduced new hopes for the treatment of serious diseases. But this promise has raised many complex questions. This field causes debate and challenge, not only among scientists but also among ethicists, religious scholars, governments, and politicians. There is no consensus on the morality of human cloning, even within specific religious traditions. In countries in which religion has a strong influence on political decision making, the moral status of the human embryo is at the center of the debate. Because of the inevitable consequences of reproductive cloning, it is prohibited in Islam. However, stem cell research for therapeutic purposes is permissible with full consideration, and all possible precautions in the pre-ensoulment stages of early fetus development, if the source is legitimate.

  11. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2010-01-01

    , we discuss how model clones arise by analyzing several practical scenarios. We propose a formal definition of models and clones, that allows us to specify a generic clone detection algorithm. Through a thorough analysis of the detail structure of sample UML domain models, recommendations for clone...

  12. 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.

  13. Advances and applications of molecular cloning in clinical microbiology.

    Science.gov (United States)

    Sharma, Kamal; Mishra, Ajay Kumar; Mehraj, Vikram; Duraisamy, Ganesh Selvaraj

    2014-10-01

    Molecular cloning is based on isolation of a DNA sequence of interest to obtain multiple copies of it in vitro. Application of this technique has become an increasingly important tool in clinical microbiology due to its simplicity, cost effectiveness, rapidity, and reliability. This review entails the recent advances in molecular cloning and its application in the clinical microbiology in the context of polymicrobial infections, recombinant antigens, recombinant vaccines, diagnostic probes, antimicrobial peptides, and recombinant cytokines. Culture-based methods in polymicrobial infection have many limitation, which has been overcome by cloning techniques and provide gold standard technique. Recombinant antigens produced by cloning technique are now being used for screening of HIV, HCV, HBV, CMV, Treponema pallidum, and other clinical infectious agents. Recombinant vaccines for hepatitis B, cholera, influenza A, and other diseases also use recombinant antigens which have replaced the use of live vaccines and thus reduce the risk for adverse effects. Gene probes developed by gene cloning have many applications including in early diagnosis of hereditary diseases, forensic investigations, and routine diagnosis. Industrial application of this technology produces new antibiotics in the form of antimicrobial peptides and recombinant cytokines that can be used as therapeutic agents.

  14. Entangled cloning of stabilizer codes and free fermions

    Science.gov (United States)

    Hsieh, Timothy H.

    2016-10-01

    Though the no-cloning theorem [Wooters and Zurek, Nature (London) 299, 802 (1982), 10.1038/299802a0] prohibits exact replication of arbitrary quantum states, there are many instances in quantum information processing and entanglement measurement in which a weaker form of cloning may be useful. Here, I provide a construction for generating an "entangled clone" for a particular but rather expansive and rich class of states. Given a stabilizer code or free fermion Hamiltonian, this construction generates an exact entangled clone of the original ground state, in the sense that the entanglement between the original and the exact copy can be tuned to be arbitrarily small but finite, or large, and the relation between the original and the copy can also be modified to some extent. For example, this Rapid Communication focuses on generating time-reversed copies of stabilizer codes and particle-hole transformed ground states of free fermion systems, although untransformed clones can also be generated. The protocol leverages entanglement to simulate a transformed copy of the Hamiltonian without having to physically implement it and can potentially be realized in superconducting qubits or ultracold atomic systems.

  15. Cloning of Plasmodium falciparum by single-cell sorting

    Science.gov (United States)

    Miao, Jun; Li, Xiaolian; Cui, Liwang

    2010-01-01

    Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples. PMID:20435038

  16. [Mystery and problems of cloning].

    Science.gov (United States)

    Nikitin, V A

    2010-01-01

    The attention of investigators is attracted to the fact that, in spite of great efforts in mammalian cloning, advances that have been made in this area of research are not great, and cloned animals have developmental pathologies often incompatible with life and/or reproduction ability. It is yet not clear what technical or biological factors underlie this, and how they are connected or interact with each other, which is more realistic strategically. There is a great number of articles dealing with the influence of cloning with the nuclear transfer on genetic and epigenetic reprogramming of donor cells. At the same time we can see the practical absence of analytical investigations concerning the technology of cloning as such, its weak points, and possible sources of cellular trauma in the course of microsurgery of nuclear transfer or twinning. This article discusses step by step several nuclear transfer techniques and the methods of dividing early preimplanted embryos for twinning with the aim to reveal possible sources of cell damage during micromanipulation that may have negative influence on the development of cloned organisms. Several new author's technologies based on the study of cell biophysical characteristics are described, which allow one to avoid cellular trauma during manipulation and minimize the possibility of cell damage at any rate.

  17. [Cloning and law in Hungary].

    Science.gov (United States)

    Julesz, Máté

    2015-03-01

    Reproductive human cloning is prohibited in Hungary, as in many other countries. Therapeutic human cloning is not prohibited, just like in many other countries. Stem cell therapy is also allowed. Article III, paragraph (3) of the Hungarian basic law (constitution) strictly forbids total human cloning. Article 1 of the Additional Protocol to the Oviedo Convention, on the Prohibition of Cloning Human Beings (1998) stipulates that any intervention seeking to create a human being genetically identical to another human being, whether living or dead, is prohibited. In Hungary, according to Article 174 of the Criminal Code, total human cloning constitutes a crime. Article 180, paragraph (3) of the Hungarian Act on Health declares that embryos shall not be brought about for research purposes; research shall be conducted only on embryos brought about for reproductive purposes when this is authorized by the persons entitled to decide upon its disposal, or when the embryo is damaged. Article 180, paragraph (5) of the Hungarian Act on Health stipulates that multiple individuals who genetically conform to one another shall not be brought about. According to Article 181, paragraph (1) of the Hungarian Act on Health, an embryo used for research shall be kept alive for not longer than 14 days, not counting the time it was frozen for storage and the time period of research.

  18. Public perceptions of animal cloning

    DEFF Research Database (Denmark)

    Jelsøe, Erling; Vincentsen, Ulla; Andersen, Ida-Elisabeth

    What was from the outset meant to be a survey testing predefined categories of ethical positions related to new biotechnologies with animal cloning as an example was subsequently developed into a process of broader involvement of groups of citizens in the issue. The survey was conducted at meetings...... in four different cities in Denmark. The participants were introduced to animal cloning and after that they filled out the questionnaire. Finally, the issue was discussed in focus groups. The process as a whole was run in a dialogue oriented way. Through the information they received in combination...... with reflecting on the survey questions the participants were well prepared for discussions in the focus groups. This approach made it possible, on the one hand to get a measure of the citizen's perceptions of the ethical aspects of animal cloning, but also to go deeper into their own thoughts of the issue...

  19. Rapid growth of a Eurasian haplotype of Phragmites australis in a restored brackish marsh in Louisiana, USA

    Science.gov (United States)

    Howard, R.J.; Travis, S.E.; Sikes, B.A.

    2008-01-01

    While numerous studies have documented patterns of invasion by non-indigenous plant species, few have considered the invasive properties of non-native genotypes of native species. Characteristics associated with specific genotypes, such as tolerance to disturbance, may mistakenly be applied to an entire species in the absence of genetic information, which consequently may affect management decisions. We report here on the incidence and growth of an introduced lineage of Phragmites australis in the Gulf of Mexico coastal zone of Louisiana. P. australis was collected from nine separate locations for inclusion in a series of growth experiments. Chloroplast DNA analysis indicated that specimens collected from four locations in the Mississippi River Delta represented the introduced Eurasian haplotype; the remainder represented the gulf coast haplotype. Three distinct genotypes, or clones, were identified within each haplotype via analysis using amplified fragment length polymorphisms, which also revealed reduced genetic diversity of the gulf coast clones compared to the Eurasian clones. Clones of each haplotype were planted along with three other native macrophytes at similar densities in a restored brackish marsh and monitored for growth. After 14 months, the Eurasian haplotype had spread vegetatively to cover about 82% of the experimental plots, more than four times the coverage (18%) of the gulf coast haplotype. Thus, the use of P. australis plantings for wetland restoration should consider the genetic lineage of plants used since our results indicate the potential of the Eurasian haplotype to grow rapidly at newly restored sites. This rapid growth may limit the establishment of more slowly growing native species. ?? 2007 Springer Science+Business Media B.V.

  20. The topsy-turvy cloning law.

    Science.gov (United States)

    Brassington, Iain; Oultram, Stuart

    2011-03-01

    In debates about human cloning, a distinction is frequently drawn between therapeutic and reproductive uses of the technology. Naturally enough, this distinction influences the way that the law is framed. The general consensus is that therapeutic cloning is less morally problematic than reproductive cloning--one can hold this position while holding that both are morally unacceptable--and the law frequently leaves the way open for some cloning for the sake of research into new therapeutic techniques while banning it for reproductive purposes. We claim that the position adopted by the law has things the wrong way around: if we accept a moral distinction between therapeutic and reproductive cloning, there are actually more reasons to be morally worried about therapeutic cloning than about reproductive cloning. If cloning is the proper object of legal scrutiny, then, we ought to make sure that we are scrutinising the right kind of clone.

  1. An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy

    OpenAIRE

    Bi Yanzhen; Qiao Xianfeng; Hua Zaidong; Zhang Liping; Liu Ximei; Li Li; Hua Wenjun; Xiao Hongwei; Zhou Jingrong; Wei Qingxin; Zheng Xinmin

    2012-01-01

    Abstract Background Widely used restriction-dependent cloning methods are labour-intensive and time-consuming, while several types of ligase-independent cloning approaches have inherent limitations. A rapid and reliable method of cloning native DNA sequences into desired plasmids are highly desired. Results This paper introduces ABI-REC, a novel strategy combining asymmetric bridge PCR with intramolecular homologous recombination in bacteria for native DNA cloning. ABI-REC was developed to pr...

  2. 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.

  3. Cloning and characterization of a novel cysteine protease gene ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    The amplified product was purified (Tiangen,. China) and cloned into the pGEM-T easy vector (Promega,. USA) followed by sequencing. 2.4 3′-RACE of HbCP1. The 3′-ready cDNA was synthesized by reversely transcribing 1 μg total RNA with 3′-CDS primer A (provided in the kit). The 3′-rapid amplification of cDNA ...

  4. 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

  5. Positional cloning of deafness genes

    NARCIS (Netherlands)

    Kremer, H.; Cremers, F.P.M.

    2009-01-01

    The identification of the majority of the known causative genes involved in nonsyndromic sensorineural hearing loss (NSHL) started with linkage analysis as part of a positional cloning procedure. The human and mouse genome projects in combination with technical developments on genotyping,

  6. Quantum cloning machines and the applications

    Science.gov (United States)

    Fan, Heng; Wang, Yi-Nan; Jing, Li; Yue, Jie-Dong; Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu

    2014-11-01

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the security of quantum key distribution protocols such as BB84 protocol, six-state protocol, B92 protocol and their generalizations. Some well-known quantum cloning machines include universal quantum cloning machine, phase-covariant cloning machine, the asymmetric quantum cloning machine and the probabilistic quantum cloning machine. In the past years, much progress has been made in studying quantum cloning machines and their applications and implementations, both theoretically and experimentally. In this review, we will give a complete description of those important developments about quantum cloning and some related topics. On the other hand, this review is self-consistent, and in particular, we try to present some detailed formulations so that further study can be taken based on those results.

  7. Quantum cloning machines and the applications

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Heng, E-mail: hfan@iphy.ac.cn [Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190 (China); Collaborative Innovation Center of Quantum Matter, Beijing 100190 (China); Wang, Yi-Nan; Jing, Li [School of Physics, Peking University, Beijing 100871 (China); Yue, Jie-Dong [Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190 (China); Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu [School of Physics, Peking University, Beijing 100871 (China)

    2014-11-20

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the security of quantum key distribution protocols such as BB84 protocol, six-state protocol, B92 protocol and their generalizations. Some well-known quantum cloning machines include universal quantum cloning machine, phase-covariant cloning machine, the asymmetric quantum cloning machine and the probabilistic quantum cloning machine. In the past years, much progress has been made in studying quantum cloning machines and their applications and implementations, both theoretically and experimentally. In this review, we will give a complete description of those important developments about quantum cloning and some related topics. On the other hand, this review is self-consistent, and in particular, we try to present some detailed formulations so that further study can be taken based on those results.

  8. Energy Values of Nine Populus Clones

    Science.gov (United States)

    Terry F. Strong

    1980-01-01

    Compares calorific values for components of nine Populus clones. The components include stem wood, stem bark, and branches. Also compares calorific values for clones of balsam poplar and black cottonwood parentages.

  9. Human reproductive cloning: a conflict of liberties.

    Science.gov (United States)

    Havstad, Joyce C

    2010-02-01

    Proponents of human reproductive cloning do not dispute that cloning may lead to violations of clones' right to self-determination, or that these violations could cause psychological harms. But they proceed with their endorsement of human reproductive cloning by dismissing these psychological harms, mainly in two ways. The first tactic is to point out that to commit the genetic fallacy is indeed a mistake; the second is to invoke Parfit's non-identity problem. The argument of this paper is that neither approach succeeds in removing our moral responsibility to consider and to prevent psychological harms to cloned individuals. In fact, the same commitment to personal liberty that generates the right to reproduce by means of cloning also creates the need to limit that right appropriately. Discussion of human reproductive cloning ought to involve a careful and balanced consideration of both the relevant aspects of personal liberty - the parents' right to reproductive freedom and the cloned child's right to self-determination.

  10. A One Pot, One Step, Precision Cloning Method with High Throughput Capability

    OpenAIRE

    Carola Engler; Romy Kandzia; Sylvestre Marillonnet

    2008-01-01

    Current cloning technologies based on site-specific recombination are efficient, simple to use, and flexible, but have the drawback of leaving recombination site sequences in the final construct, adding an extra 8 to 13 amino acids to the expressed protein. We have devised a simple and rapid subcloning strategy to transfer any DNA fragment of interest from an entry clone into an expression vector, without this shortcoming. The strategy is based on the use of type IIs restriction enzymes, whic...

  11. Cloning of the repertoire of individual Plasmodium falciparum var genes using transformation associated recombination (TAR.

    Directory of Open Access Journals (Sweden)

    Annette Gaida

    Full Text Available One of the major virulence factors of the malaria causing parasite is the Plasmodium falciparum encoded erythrocyte membrane protein 1 (PfEMP1. It is translocated to It the membrane of infected erythrocytes and expressed from approximately 60 var genes in a mutually exclusive manner. Switching of var genes allows the parasite to alter functional and antigenic properties of infected erythrocytes, to escape the immune defense and to establish chronic infections. We have developed an efficient method for isolating VAR genes from telomeric and other genome locations by adapting transformation-associated recombination (TAR cloning, which can then be analyzed and sequenced. For this purpose, three plasmids each containing a homologous sequence representing the upstream regions of the group A, B, and C var genes and a sequence homologous to the conserved acidic terminal segment (ATS of var genes were generated. Co-transfection with P. falciparum strain ITG2F6 genomic DNA in yeast cells yielded 200 TAR clones. The relative frequencies of clones from each group were not biased. Clones were screened by PCR, as well as Southern blotting, which revealed clones missed by PCR due to sequence mismatches with the primers. Selected clones were transformed into E. coli and further analyzed by RFLP and end sequencing. Physical analysis of 36 clones revealed 27 distinct types potentially representing 50% of the var gene repertoire. Three clones were selected for sequencing and assembled into single var gene containing contigs. This study demonstrates that it is possible to rapidly obtain the repertoire of var genes from P. falciparum within a single set of cloning experiments. This technique can be applied to individual isolates which will provide a detailed picture of the diversity of var genes in the field. This is a powerful tool to overcome the obstacles with cloning and assembly of multi-gene families by simultaneously cloning each member.

  12. Cloning of the repertoire of individual Plasmodium falciparum var genes using transformation associated recombination (TAR).

    Science.gov (United States)

    Gaida, Annette; Becker, Marion M; Schmid, Christoph D; Bühlmann, Tobias; Louis, Edward J; Beck, Hans-Peter

    2011-03-07

    One of the major virulence factors of the malaria causing parasite is the Plasmodium falciparum encoded erythrocyte membrane protein 1 (PfEMP1). It is translocated to It the membrane of infected erythrocytes and expressed from approximately 60 var genes in a mutually exclusive manner. Switching of var genes allows the parasite to alter functional and antigenic properties of infected erythrocytes, to escape the immune defense and to establish chronic infections. We have developed an efficient method for isolating VAR genes from telomeric and other genome locations by adapting transformation-associated recombination (TAR) cloning, which can then be analyzed and sequenced. For this purpose, three plasmids each containing a homologous sequence representing the upstream regions of the group A, B, and C var genes and a sequence homologous to the conserved acidic terminal segment (ATS) of var genes were generated. Co-transfection with P. falciparum strain ITG2F6 genomic DNA in yeast cells yielded 200 TAR clones. The relative frequencies of clones from each group were not biased. Clones were screened by PCR, as well as Southern blotting, which revealed clones missed by PCR due to sequence mismatches with the primers. Selected clones were transformed into E. coli and further analyzed by RFLP and end sequencing. Physical analysis of 36 clones revealed 27 distinct types potentially representing 50% of the var gene repertoire. Three clones were selected for sequencing and assembled into single var gene containing contigs. This study demonstrates that it is possible to rapidly obtain the repertoire of var genes from P. falciparum within a single set of cloning experiments. This technique can be applied to individual isolates which will provide a detailed picture of the diversity of var genes in the field. This is a powerful tool to overcome the obstacles with cloning and assembly of multi-gene families by simultaneously cloning each member.

  13. Effective and efficient model clone detection

    DEFF Research Database (Denmark)

    Störrle, Harald

    2015-01-01

    automatically (“MDD-style”) or hand-crafted following the blueprint defined by the model (“MBSD-style”). Unfortunately, however, model clones are much less well studied than code clones. In this paper, we present a clone detection algorithm for UML domain models. Our approach covers a much greater variety...

  14. Soil water dynamics and evapotranspiration of forage cactus clones under rainfed conditions

    Directory of Open Access Journals (Sweden)

    Thieres George Freire da Silva

    2015-07-01

    Full Text Available Abstract: The objective of this work was to evaluate soil water dynamics in areas cultivated with forage cactus clones and to determine how environmental conditions and crop growth affect evapotranspiration. The study was conducted in the municipality of Serra Talhada, in the state of Pernambuco, Brazil. Crop growth was monitored through changes in the cladode area index (CAI and through the soil cover fraction, calculated at the end of the cycle. Real evapotranspiration (ET of the three evaluated clones was obtained as the residual term in the soil water balance method. No difference was observed between soil water balance components, even though the evaluated clones were of different genus and had different CAI increments. Accumulated ET was of 1,173 mm during the 499 days of the experiment, resulting in daily average of 2.35 mm. The CAI increases the water consumption of the Orelha de Elefante Mexicana clone. In dry conditions, the water consumption of the Miúda clone responds more slowly to variation in soil water availability. The lower evolution of the CAI of the IPA Sertânia clone, during the rainy season, leads to a higher contribution of the evaporation component in ET. The atmospheric demand controls the ET of clones only when there is higher soil water availability; in this condition, the water consumption of the Miúda clone decreases more rapidly with the increase of atmospheric demand.

  15. A single-copy galK promoter cloning vector suitable for cloning strong promoters

    DEFF Research Database (Denmark)

    Dandanell, Gert; Court, Donald L.; Hammer, Karin

    1986-01-01

    We report the construction of lambda galK promoter cloning vectors for cloning and characterization of strong promoters. This phage, which contains a unique HindIII cloning site, was applied to the cloning and analysis of transcription initiations of the regulatory region of the deo-operon of...

  16. El envejecimiento de los clones

    OpenAIRE

    Trippi, Victorio S.

    2007-01-01

    El envejecimiento de los clones se observa en plantas que muestran crecimiento definido por un determinismo genético, cuando se multiplican con tejidos que evolucionan hacia el crecimiento reproductivo. Las plantas fuertemente influenciadas por el ambiente, pueden mostrar fenómenos de senescencia cuando la condición de ambiente determina el crecimiento reproductivo. Los cambios asociados con la edad resultan de alteraciones del citoplasma como un tipo de diferenciación cel...

  17. Therapeutic and reproductive cloning: a critique.

    Science.gov (United States)

    Bowring, Finn

    2004-01-01

    This article is a critical examination of the science and ethics of human cloning. It summarises the key scientific milestones in the development of nuclear transplantation, explains the importance of cloning to research into the medical potential of embryonic stem cells, and discusses the well-worn distinction between 'therapeutic' and 'reproductive' cloning. Suggesting that this distinction will be impossible to police, it goes on to consider the ethics of full human cloning. It is concluded that it represents an unacceptable form of parental despotism, and that the genetic engineering and cloning of future human beings will fracture the foundations of modern humanism.

  18. Cloning expeditions: risky but rewarding.

    Science.gov (United States)

    Lodish, Harvey

    2013-12-01

    In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine.

  19. Cloning Expeditions: Risky but Rewarding

    Science.gov (United States)

    2013-01-01

    In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine. PMID:24061478

  20. Effects of Planting Density on Transpiration, Stem Flow and Interception for Two Clones Differing in Drought Tolerance in a High Productivity Eucalyptus Plantation in Brazil

    Science.gov (United States)

    Hubbard, R. M.; Hakemada, R.; Ferraz, S.

    2015-12-01

    Eucalypt plantations cover about 20 M hectares worldwide and expansion is expected to mainly occur in marginal growing areas where dry conditions may lead to water conflicts. One of the principal reasons for the expansion of Eucalyptus plantations is rapid wood growth but these forests also transpire large amounts of water. Genotype selection and planting density, are key factors regulating carbon and water tradeoffs at a stand scale, but few studies have examined these simultaneously especially in highly productive clonal plantations. Our goal in this study was to examine the effects of planting density on carbon and water interactions using a drought tolerant and drought sensitive eucalyptus clone. This work is part of a larger study (TECHS project - Tolerance of Eucalyptus Clones to Hydric and Thermal Stresses) and is located in a flat Oxisol in southeast of Brazil. A drought tolerant (E. grandis x E. camaldulensis (Grancam) and drought sensitive clone E. grandis x E. urophylla (Urograndis) were planted at four densities ranging from 600 to 3.000 stem ha-1. We measured transpiration using thermal heat dissipation probes, wood growth, canopy interception and stemflow during a full year (21 to 33 months old). Precipitation during the study period was 738 mm. Independently of genetics, growth increased with increasing density. Transpiration also increased with planting density and ranged from 515-595 mm at wider spacing to 735-978 mm at tighter spacing. Interception increased with planting density representing 18-22% of precipitation versus 13-14% in wider spacing while stem flow represented 2-5% in denser spacing and 1-2% at broader spacing. When density was higher than 1.250 and 1.750 stems ha-1 in Urograndis and Grancam clones, respectively, the water balance were negative. On a stand scale, results show both genetics and spacing can be used as silvicultural tools to better manage the tradeoff between wood growth and water consumption.

  1. pClone: Synthetic Biology Tool Makes Promoter Research Accessible to Beginning Biology Students

    Science.gov (United States)

    Eckdahl, Todd; Cronk, Brian; Andresen, Corinne; Frederick, Paul; Huckuntod, Samantha; Shinneman, Claire; Wacker, Annie; Yuan, Jason

    2014-01-01

    The Vision and Change report recommended genuine research experiences for undergraduate biology students. Authentic research improves science education, increases the number of scientifically literate citizens, and encourages students to pursue research. Synthetic biology is well suited for undergraduate research and is a growing area of science. We developed a laboratory module called pClone that empowers students to use advances in molecular cloning methods to discover new promoters for use by synthetic biologists. Our educational goals are consistent with Vision and Change and emphasize core concepts and competencies. pClone is a family of three plasmids that students use to clone a new transcriptional promoter or mutate a canonical promoter and measure promoter activity in Escherichia coli. We also developed the Registry of Functional Promoters, an open-access database of student promoter research results. Using pre- and posttests, we measured significant learning gains among students using pClone in introductory biology and genetics classes. Student posttest scores were significantly better than scores of students who did not use pClone. pClone is an easy and affordable mechanism for large-enrollment labs to meet the high standards of Vision and Change. PMID:26086659

  2. Recent Achievement in Gene Cloning and Functional Genomics in Soybean

    Directory of Open Access Journals (Sweden)

    Zhengjun Xia

    2013-01-01

    Full Text Available Soybean is a model plant for photoperiodism as well as for symbiotic nitrogen fixation. However, a rather low efficiency in soybean transformation hampers functional analysis of genes isolated from soybean. In comparison, rapid development and progress in flowering time and photoperiodic response have been achieved in Arabidopsis and rice. As the soybean genomic information has been released since 2008, gene cloning and functional genomic studies have been revived as indicated by successfully characterizing genes involved in maturity and nematode resistance. Here, we review some major achievements in the cloning of some important genes and some specific features at genetic or genomic levels revealed by the analysis of functional genomics of soybean.

  3. Future issues in transplantation ethics: ethical and legal controversies in xenotransplantation, stem cell, and cloning research.

    Science.gov (United States)

    Shapiro, Robyn S

    2008-07-01

    With little prospect of developing a sufficient supply of human transplantable organs to meet the large and growing demand, attention has turned to xenotransplantation, as well as stem cell and cloning research, as possible approaches for alleviating this allograft shortage. This article explores ethical and legal issues that surround developments in these fields.

  4. Successful pod infections by Moniliophthora roreri result in differential Theobroma cacao gene expression depending on the clone's level of tolerance.

    Science.gov (United States)

    Ali, Shahin S; Melnick, Rachel L; Crozier, Jayne; Phillips-Mora, Wilberth; Strem, Mary D; Shao, Jonathan; Zhang, Dapeng; Sicher, Richard; Meinhardt, Lyndel; Bailey, Bryan A

    2014-09-01

    An understanding of the tolerance mechanisms of Theobroma cacao used against Moniliophthora roreri, the causal agent of frosty pod rot, is important for the generation of stable disease-tolerant clones. A comparative view was obtained of transcript populations of infected pods from two susceptible and two tolerant clones using RNA sequence (RNA-Seq) analysis. A total of 3009 transcripts showed differential expression among clones. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of differentially expressed genes indicated shifts in 152 different metabolic pathways between the tolerant and susceptible clones. Real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) analyses of 36 genes verified the differential expression. Regression analysis validated a uniform progression in gene expression in association with infection levels and fungal loads in the susceptible clones. Expression patterns observed in the susceptible clones diverged in tolerant clones, with many genes showing higher expression at a low level of infection and fungal load. Principal coordinate analyses of real-time qRT-PCR data separated the gene expression patterns between susceptible and tolerant clones for pods showing malformation. Although some genes were constitutively differentially expressed between clones, most results suggested that defence responses were induced at low fungal load in the tolerant clones. Several elicitor-responsive genes were highly expressed in tolerant clones, suggesting rapid recognition of the pathogen and induction of defence genes. Expression patterns suggested that the jasmonic acid-ethylene- and/or salicylic acid-mediated defence pathways were activated in the tolerant clones, being enhanced by reduced brassinosteroid (BR) biosynthesis and catabolic inactivation of both BR and abscisic acids. Finally, several genes associated with hypersensitive response-like cell death were also induced in tolerant clones. © 2014

  5. Hybrid sequencing approach applied to human fecal metagenomic clone libraries revealed clones with potential biotechnological applications.

    Directory of Open Access Journals (Sweden)

    Mária Džunková

    Full Text Available Natural environments represent an incredible source of microbial genetic diversity. Discovery of novel biomolecules involves biotechnological methods that often require the design and implementation of biochemical assays to screen clone libraries. However, when an assay is applied to thousands of clones, one may eventually end up with very few positive clones which, in most of the cases, have to be "domesticated" for downstream characterization and application, and this makes screening both laborious and expensive. The negative clones, which are not considered by the selected assay, may also have biotechnological potential; however, unfortunately they would remain unexplored. Knowledge of the clone sequences provides important clues about potential biotechnological application of the clones in the library; however, the sequencing of clones one-by-one would be very time-consuming and expensive. In this study, we characterized the first metagenomic clone library from the feces of a healthy human volunteer, using a method based on 454 pyrosequencing coupled with a clone-by-clone Sanger end-sequencing. Instead of whole individual clone sequencing, we sequenced 358 clones in a pool. The medium-large insert (7-15 kb cloning strategy allowed us to assemble these clones correctly, and to assign the clone ends to maintain the link between the position of a living clone in the library and the annotated contig from the 454 assembly. Finally, we found several open reading frames (ORFs with previously described potential medical application. The proposed approach allows planning ad-hoc biochemical assays for the clones of interest, and the appropriate sub-cloning strategy for gene expression in suitable vectors/hosts.

  6. Homologous cloning, characterization and expression of a new halophyte phytochelatin synthase gene in Suaeda salsa

    Science.gov (United States)

    Cong, Ming; Zhao, Jianmin; Lü, Jiasen; Ren, Zhiming; Wu, Huifeng

    2016-09-01

    The halophyte Suaeda salsa can grow in heavy metal-polluted areas along intertidal zones having high salinity. Since phytochelatins can eff ectively chelate heavy metals, it was hypothesized that S. salsa possessed a phytochelatin synthase (PCS) gene. In the present study, the cDNA of PCS was obtained from S. salsa (designated as SsPCS) using homologous cloning and the rapid amplification of cDNA ends (RACE). A sequence analysis revealed that SsPCS consisted of 1 916 bp nucleotides, encoding a polypeptide of 492 amino acids with one phytochelatin domain and one phytochelatin C domain. A similarity analysis suggested that SsPCS shared up to a 58.6% identity with other PCS proteins and clustered with PCS proteins from eudicots. There was a new kind of metal ion sensor motif in its C-terminal domain. The SsPCS transcript was more highly expressed in elongated and fibered roots and stems ( Pcloned from a halophyte, and it might contain a diff erent metal sensing capability than the first PCS from Thellungiella halophila. This study provided a new view of halophyte PCS genes in heavy metal tolerance.

  7. Growing Bacteriophage M13 in Liquid Culture.

    Science.gov (United States)

    Green, Michael R; Sambrook, Joseph

    2017-11-01

    Stocks of bacteriophage M13 are usually grown in liquid culture. The infected bacteria do not lyse but, instead, grow at a slower than normal rate to form a dilute suspension. The inoculum of bacteriophage is almost always a freshly picked plaque or a suspension of bacteriophage particles obtained from a single plaque, as described here. Infected cells contain up to 200 copies of double-stranded, replicative-form DNA and extrude several hundred bacteriophage particles per generation. Thus, a 1-mL culture of infected cells can produce enough double-stranded viral DNA (1-2 mg) for restriction mapping and recovery of cloned DNA inserts and sufficient single-stranded DNA (∼5-10 mg) for site-directed mutagenesis, DNA sequencing, or synthesis of radiolabeled probes. The titer of bacteriophages in the supernatant from infected cells is so high (∼10(12) pfu/mL) that a small aliquot serves as a permanent stock of the starting plaque. © 2017 Cold Spring Harbor Laboratory Press.

  8. Cloning and assisted reproductive techniques: influence on early development and adult phenotype.

    Science.gov (United States)

    Sakai, Randall R; Tamashiro, Kellie L K; Yamazaki, Yukiko; Yanagimachi, Ryuzo

    2005-06-01

    Over the past 40 years, our increased understanding and development of cell and molecular biology has allowed even greater advances in reproductive biology. This is most evident by the development of various aspects of assisted reproductive techniques (ART), generation of transgenic animals, and most recently generation of mammals through somatic cell cloning. To date, cloning from adult somatic cells has been successful in at least 10 mammalian species. Although generating viable cloned mammals from adult cells is technically feasible and the list of successes will only continue to grow with time, prenatal and perinatal mortality is high and live cloned offspring have not been without health problems. The success of many of the proposed applications of the cloning technique obviously depends upon the health and survival of founder animals generated by nuclear transfer. This article summarizes the health consequences of cloning in mice, and discusses possible mechanisms through which these conditions may arise. In addition, we discuss the effects of ART in animal models and in humans. ART also involves some of the same procedures used in cloning, and there are reports that offspring generated by ART sometimes display aberrant phenotypes as well. It is important to point out that although these techniques do sometimes produce abnormalities, the majority of offspring are born apparently normal and survive to adulthood. Additionally, we must emphasize that the effects of ART and cloning observed in animal models do not necessarily indicate that they will occur in humans. In this article, we review studies examining the phenotype of animals generated by cloning and various ART, and discuss clinical implications of these findings. (c) 2005 Wiley-Liss, Inc.

  9. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2013-01-01

    Code clones (i.e., duplicate fragments of code) have been studied for long, and there is strong evidence that they are a major source of software faults. Anecdotal evidence suggests that this phenomenon occurs similarly in models, suggesting that model clones are as detrimental to model quality...... as they are to code quality. However, programming language code and visual models have significant differences that make it difficult to directly transfer notions and algorithms developed in the code clone arena to model clones. In this article, we develop and propose a definition of the notion of “model clone” based...... on the thorough analysis of practical scenarios. We propose a formal definition of model clones, specify a clone detection algorithm for UML domain models, and implement it prototypically. We investigate different similarity heuristics to be used in the algorithm, and report the performance of our approach. While...

  10. Cloning cattle: the methods in the madness.

    Science.gov (United States)

    Oback, Björn; Wells, David N

    2007-01-01

    Somatic cell nuclear transfer (SCNT) is much more widely and efficiently practiced in cattle than in any other species, making this arguably the most important mammal cloned to date. While the initial objective behind cattle cloning was commercially driven--in particular to multiply genetically superior animals with desired phenotypic traits and to produce genetically modified animals-researchers have now started to use bovine SCNT as a tool to address diverse questions in developmental and cell biology. In this paper, we review current cattle cloning methodologies and their potential technical or biological pitfalls at any step of the procedure. In doing so, we focus on one methodological parameter, namely donor cell selection. We emphasize the impact of epigenetic and genetic differences between embryonic, germ, and somatic donor cell types on cloning efficiency. Lastly, we discuss adult phenotypes and fitness of cloned cattle and their offspring and illustrate some of the more imminent commercial cattle cloning applications.

  11. High-throughput clone screening followed by protein expression cross-check: A visual assay platform.

    Science.gov (United States)

    Bose, Partha Pratim; Kumar, Prakash

    2017-01-01

    In high-throughput biotechnology and structural biology, molecular cloning is an essential prerequisite for attaining high yields of recombinant protein. However, a rapid, cost-effective, easy clone screening protocol is still required to identify colonies with desired insert along with a cross check method to certify the expression of the desired protein as the end product. We report an easy, fast, sensitive and cheap visual clone screening and protein expression cross check protocol employing gold nanoparticle based plasmonic detection phenomenon. This is a non-gel, non-PCR based visual detection technique, which can be used as simultaneous high throughput clone screening followed by the determination of expression of desired protein. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. 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.

  13. AN APPROACH FOR CLONE DETECTION IN DOCUMENTATION REUSE

    OpenAIRE

    D. V. Lutsiv; D. V. Koznov; H. A. Basit; E. L. Ouh; M. N. Smirnov; K. Y. Romanovsky

    2014-01-01

    The paper focuses on the searching method for repetitions in DocBook/DRL or plain text documents. An algorithm has been designed based on software clone detection. The algorithm supports filtering results: clones are rejected if clone length in the group is less than 5 symbols, intersection of clone groups is eliminated, meaningfulness clones are removed, the groups containing clones consisting only of XML are eliminated. Remaining search is supported: found clones are extracted from the docu...

  14. Human cloning: Eastern Mediterranean Region perspective.

    Science.gov (United States)

    Abdur Rab, M; Khayat, M H

    2006-01-01

    Recent advances in genomics and biotechnology have ushered in a new era in health development. Therapeutic cloning possesses enormous potential for revolutionizing medical and therapeutic techniques. Cloning technology, however, is perceived as having the potential for reproductive cloning, which raises serious ethical and moral concerns. It is important that the Islamic countries come to a consensus on this vital issue. Developing science and technology for better health is a religious and moral obligation. There is an urgent need for Muslim scholars to discuss the issue of stem cell research and cloning rationally; such dialogue will not only consider the scientific merits but also the moral, ethical and legal implications.

  15. Metabolomic phenotyping of a cloned pig model

    Directory of Open Access Journals (Sweden)

    Callesen Henrik

    2011-08-01

    Full Text Available Abstract Background Pigs are widely used as models for human physiological changes in intervention studies, because of the close resemblance between human and porcine physiology and the high degree of experimental control when using an animal model. Cloned animals have, in principle, identical genotypes and possibly also phenotypes and this offer an extra level of experimental control which could possibly make them a desirable tool for intervention studies. Therefore, in the present study, we address how phenotype and phenotypic variation is affected by cloning, through comparison of cloned pigs and normal outbred pigs. Results The metabolic phenotype of cloned pigs (n = 5 was for the first time elucidated by nuclear magnetic resonance (NMR-based metabolomic analysis of multiple bio-fluids including plasma, bile and urine. The metabolic phenotype of the cloned pigs was compared with normal outbred pigs (n = 6 by multivariate data analysis, which revealed differences in the metabolic phenotypes. Plasma lactate was higher for cloned vs control pigs, while multiple metabolites were altered in the bile. However a lower inter-individual variability for cloned pigs compared with control pigs could not be established. Conclusions From the present study we conclude that cloned and normal outbred pigs are phenotypically different. However, it cannot be concluded that the use of cloned animals will reduce the inter-individual variation in intervention studies, though this is based on a limited number of animals.

  16. Metabolomic phenotyping of a cloned pig model

    Science.gov (United States)

    2011-01-01

    Background Pigs are widely used as models for human physiological changes in intervention studies, because of the close resemblance between human and porcine physiology and the high degree of experimental control when using an animal model. Cloned animals have, in principle, identical genotypes and possibly also phenotypes and this offer an extra level of experimental control which could possibly make them a desirable tool for intervention studies. Therefore, in the present study, we address how phenotype and phenotypic variation is affected by cloning, through comparison of cloned pigs and normal outbred pigs. Results The metabolic phenotype of cloned pigs (n = 5) was for the first time elucidated by nuclear magnetic resonance (NMR)-based metabolomic analysis of multiple bio-fluids including plasma, bile and urine. The metabolic phenotype of the cloned pigs was compared with normal outbred pigs (n = 6) by multivariate data analysis, which revealed differences in the metabolic phenotypes. Plasma lactate was higher for cloned vs control pigs, while multiple metabolites were altered in the bile. However a lower inter-individual variability for cloned pigs compared with control pigs could not be established. Conclusions From the present study we conclude that cloned and normal outbred pigs are phenotypically different. However, it cannot be concluded that the use of cloned animals will reduce the inter-individual variation in intervention studies, though this is based on a limited number of animals. PMID:21859467

  17. Species-specific challenges in dog cloning.

    Science.gov (United States)

    Kim, G A; Oh, H J; Park, J E; Kim, M J; Park, E J; Jo, Y K; Jang, G; Kim, M K; Kim, H J; Lee, B C

    2012-12-01

    Somatic cell nuclear transfer (SCNT) is now an established procedure used in cloning of several species. SCNT in dogs involves multiple steps including the removal of the nuclear material, injection of a donor cell, fusion, activation of the reconstructed oocytes and finally transfer to a synchronized female recipient. There are therefore many factors that contribute to cloning efficiency. By performing a retrospective analysis of 2005-2012 published papers regarding dog cloning, we define the optimum procedure and summarize the specific feature for dog cloning. © 2012 Blackwell Verlag GmbH.

  18. Quantum cloning disturbed by thermal Davies environment

    Science.gov (United States)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-06-01

    A network of quantum gates designed to implement universal quantum cloning machine is studied. We analyze how thermal environment coupled to auxiliary qubits, `blank paper' and `toner' required at the preparation stage of copying, modifies an output fidelity of the cloner. Thermal environment is described in terms of the Markovian Davies theory. We show that such a cloning machine is not universal any more but its output is independent of at least a part of parameters of the environment. As a case study, we consider cloning of states in a six-state cryptography's protocol. We also briefly discuss cloning of arbitrary input states.

  19. Human cloning and our sense of self.

    Science.gov (United States)

    Brock, Dan W

    2002-04-12

    This Viewpoint formulates and responds to three lines of argument concerning human reproductive cloning's potential to undermine our sense of self or identity. First, cloning would undermine our sense of individuality or uniqueness. But it could only undermine our genetic uniqueness, not our full individuality. Second, cloning would undermine the value or worth of human beings. But it would not make individuals replaceable or of any less moral worth. Third, a clone's freedom or autonomy to construct his or her own life would be undermined by the presence of an earlier twin. But only a mistaken belief in genetic determinism supports this feared loss of freedom.

  20. Biomass production of four willow clones grown as short rotation coppice on two soil types in Denmark

    DEFF Research Database (Denmark)

    Sevel, Lisbeth; Nord-Larsen, Thomas; Raulund-Rasmussen, Karsten

    2012-01-01

    Ambitious targets for reducing emissions of carbon dioxide have created a demand for renewable sources of energy. Short rotation coppice (SRC) willow has the potential for meeting part of this demand. In this study, an experiment including four commercial clones of willow grown on two different......-destructive method showed a large increase in annual biomass production from the first to the second growing season. Based on the harvested willow, average annual biomass production of the four clones ranged from 5.2 to 8.8 odt ha-1 yr-1 with a significant effect of both soil type and clone. The interaction between...

  1. Homologous recombination-mediated cloning and manipulation of genomic DNA regions using Gateway and recombineering systems.

    Science.gov (United States)

    Rozwadowski, Kevin; Yang, Wen; Kagale, Sateesh

    2008-11-17

    Employing genomic DNA clones to characterise gene attributes has several advantages over the use of cDNA clones, including the presence of native transcription and translation regulatory sequences as well as a representation of the complete repertoire of potential splice variants encoded by the gene. However, working with genomic DNA clones has traditionally been tedious due to their large size relative to cDNA clones and the presence, absence or position of particular restriction enzyme sites that may complicate conventional in vitro cloning procedures. To enable efficient cloning and manipulation of genomic DNA fragments for the purposes of gene expression and reporter-gene studies we have combined aspects of the Gateway system and a bacteriophage-based homologous recombination (i.e. recombineering) system. To apply the method for characterising plant genes we developed novel Gateway and plant transformation vectors that are of small size and incorporate selectable markers which enable efficient identification of recombinant clones. We demonstrate that the genomic coding region of a gene can be directly cloned into a Gateway Entry vector by recombineering enabling its subsequent transfer to Gateway Expression vectors. We also demonstrate how the coding and regulatory regions of a gene can be directly cloned into a plant transformation vector by recombineering. This construct was then rapidly converted into a novel Gateway Expression vector incorporating cognate 5' and 3' regulatory regions by using recombineering to replace the intervening coding region with the Gateway Destination cassette. Such expression vectors can be applied to characterise gene regulatory regions through development of reporter-gene fusions, using the Gateway Entry clones of GUS and GFP described here, or for ectopic expression of a coding region cloned into a Gateway Entry vector. We exemplify the utility of this approach with the Arabidopsis PAP85 gene and demonstrate that the expression

  2. Multiple factors affect pest and pathogen damage on 31 Populus clones in South Carolina

    Energy Technology Data Exchange (ETDEWEB)

    Coyle, David R.; Coleman, Mark D. [USDA Forest Service, Southern Research Station, P.O. Box 700, New Ellenton, SC 29809 (United States); Durant, Jaclin A.; Newman, Lee A. [Arnold School of Public Health, Department of Environmental Health Sciences, University of South Carolina, 800 Sumter St., Columbia, SC 29208 (United States)

    2006-08-15

    Populus species and hybrids have many practical applications, but there is a paucity of data regarding selections that perform well in the southeastern US. We compared pest susceptibility of 31 Populus clones over 3 years in South Carolina, USA. Cuttings were planted in spring 2001 on two study sites. Clones planted in the bottomland site received granular fertilizer yearly and irrigation the first two years only, while those on the sandy, upland site received irrigation and fertilization throughout each growing season. Foliar damage by the cottonwood leaf beetle (Chrysomela scripta), cottonwood leafcurl mite (Tetra lobulifera), and poplar leaf rust (Melampsora medusae) was visually monitored several times each growing season. Damage ratings differed significantly among clones, and clonal rankings changed from year to year. Irrigation increased C. scripta and M. medusae damage, but had no effect on T. lobulifera damage. Certain clones received greater pest damage at a particular study site. Temporal damage patterns were evident among individual clones and on each site. At the upland site, OP367 and 7300502 were highly resistant to all three pests; I45/51 was highly resistant to C. scripta and M. medusae; NM6 and 15-29 were highly resistant to M. medusae; and 7302801 was highly resistant to T. lobulifera and M. medusae. At the bottomland site, NM6, Eridano, I45/51, and 7302801 were highly resistant to all three pests; clone 7300502 was highly resistant to M. medusae only. Based on this preliminary 3-year study of pest damage levels, we would recommend clones NM6, Eridano, I45/51, OP367, 15-29, 7302801, 7300502, and Kentucky 8 for use in this region. (author)

  3. Pattern of Flushing, Cherelle Wilt, and Accuracy of Yield Forecasting of Some Cocoa Clones

    Directory of Open Access Journals (Sweden)

    Adi Prawoto

    2014-08-01

    Full Text Available Monthly observation of cocoa flushing, number of cherelle wilt (CW, number of small, medium and large pods of 6 clones was conducted for two years to study its dynamics for one year. A study was conducted in Kaliwining Experimental Station, 45 m asl. and D rainfall type (according to Schmidt & Ferguson, using ICS 13, ICS 60, TSH 858, Sulawesi 1, Sulawesi 2 and KW 165 clones of 8 years old. Each clone was planted intermittently in separate rows, replicated 6 rows. Correlation and regression analysis were done between variables and with rainfall data. The parallel research was conducted in the similar station to assess the accuracy of production estimation method by identify percentage of small pods (length 1—10 cm, medium (11—15 cm and large pods (>15 cm to grow until harvested. The study used 15th years old trees of Sulawesi 1, Sulawesi 2, KW 165, KKM 22, ICS 13 and DR 2 clones. Each clones was replicated 5 times. The result showed that intensive flushing (>50% occured during January, March, September and November meanwhile no flushing during December and February. Correlation between rainfall and flushing was positive (r=0.27. Effect of clones on flushing frequency was similar but for flushing intensity was significant. KW 165 tended to be the lowest but TSH 858 tend to be the highest. CW occured for a year-round but the height level during May and June. Effect of clones was significant, KW 165 showed highest followed by Sulawesi 2. CW level showed positive correlation with number of medium (r=0.71 and big pods (r=0.55, except showed negative correlation with flushing intensity (r=-0.37 and rainfall (r=-0.51. High pod setting happened during May to November and low pod setting during December to March. In this aspect effect of clones were significant, the productive clones were Sulawesi 1, Sulawesi 2 and KW 165, but ICS 60 was the less. CW level during 1st semester was lower than at 2nd semester and clone effect was significant. The

  4. Future and applications of cloning.

    Science.gov (United States)

    Trounson, Alan O

    2006-01-01

    The birth of viable offspring from somatic cell nuclear transfer (SCNT) in mammals caused a major re-examination of the understanding of the commitment of cells to specific tissue lineages during differentiation. The questions of whether cells undergo dedifferentiation or transdifferentiation during the development of offspring and how these changes are controlled is a source of ongoing debate that is yet to be resolved. Irrespective of the outcome of this debate, it is clear that cloning using SCNT has a place and purpose in the future of research and animal breeding. The future uses of SCNT could include the production of transgenic mice, the production of transgenic livestock and assisting with the re-establishment of endangered species. Human medicine also would benefit from future use of SCNT because it would allow the production of patient-specific embryonic stem cells.

  5. 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

  6. [Construction of RNAi vectors for SmNAC1 transcription factors of Salvia miltiorrhiza using Gateway cloning technology].

    Science.gov (United States)

    Zhao, Rong; Rong, Qi-Xian; Liu, Yu-Zhong; Shen, Ye; Huang, Lu-Qi

    2014-05-01

    NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene.

  7. Survival and Growth of Epidemically Successful and Nonsuccessful Salmonella enterica Clones after Freezing and Dehydration

    DEFF Research Database (Denmark)

    Müller, Karoline; Aabo, Søren; Birk, Tina

    2012-01-01

    The spread of epidemically successful nontyphoidal Salmonella clones has been suggested as the most important cause of salmonellosis in industrialized countries. Factors leading to the emergence of success clones are largely unknown, but their ability to survive and grow after physical stress may...... contribute. During epidemiological studies, a mathematical model was developed that allowed estimation of a factor (q) accounting for the relative ability of Salmonella serovars with different antimicrobial resistances to survive in the food chain and cause human disease. Based on this q-factor, 26...

  8. The ethics of human reproductive cloning.

    Science.gov (United States)

    Strong, Carson

    2005-03-01

    This article addresses the question of whether human reproductive cloning could be ethically justifiable in at least some cases involving infertile couples who would choose cloning as a way to have a genetically related child. At present, the risk of congenital anomalies constitutes a compelling argument against human reproductive cloning. The article explores whether reproductive cloning could be ethically justifiable if, at some future time, cloning becomes possible without an elevated risk of anomalies. It is argued that freedom to use cloning is a form of procreative freedom and, as such, deserves respect. All of the objections that have been raised against human reproductive cloning fall under three main categories: those that appeal to the interests of the child, those based on consequences for society, and those arising from teleological views. Objections that appeal to the child's interests are, in turn, of two main kinds: consequentialist and deontological. All of these types of objections are examined, and it is found that each involves serious problems that prevent it from being a reasonable objection in the context of the infertility cases considered. It is concluded that human reproductive cloning would be ethically justifiable in at least some cases involving infertile couples, provided that it could be performed without an elevated risk of anomalies.

  9. What's so bad about human cloning?

    Science.gov (United States)

    Breitowitz, Yitzchok

    2002-12-01

    There appears to be a consensus in the general community that reproductive cloning is an immoral technology that should be banned. It may, however, be argued, at least from the perspective of the Jewish tradition, that reproductive cloning has many positive benefits. It is thus essential that one carefully weigh the costs and the benefits before deciding on a definitive course of action.

  10. Computerized adaptive testing with item cloning

    NARCIS (Netherlands)

    Glas, Cornelis A.W.; van der Linden, Willem J.

    2003-01-01

    To increase the number of items available for adaptive testing and reduce the cost of item writing, the use of techniques of item cloning has been proposed. An important consequence of item cloning is possible variability between the item parameters. To deal with this variability, a multilevel item

  11. Challenges in regulating farm animal cloning

    DEFF Research Database (Denmark)

    Gunning, Jennifer; Hartlev, Mette; Gamborg, Christian

    Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety......Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety...

  12. Cloning of partial putative gonadotropin hormone receptor ...

    Indian Academy of Sciences (India)

    Equus asinus. –. 71·0. –. Macaca fascicularis. –. 83·9. –. Gallus gallus. –. 74·2. –. Canis familiaris. –. –. 74·2. Drosophila melanogaster. –. 61·3. –. Figure 2. Structure of the cloned insert. (a) Diagrammatic representation of the cloned DNA fragment; arrangement of the PCR products are illustrated with primers. Letters 'F' and.

  13. Cloning and superluminal signaling£

    Indian Academy of Sciences (India)

    is possible, we can then have (usable) superluminal signaling [8]. The basic principle behind no exact cloning is the principle of no-signaling (i.e., no superluminal signaling). Allowing no-signaling, Gisin [7] has reproduced the 1. 2 optimal universal symmetric cloning machine of Buzek and Hillery [2] for qubits. We shall ...

  14. Metabolomic phenotyping of af cloned pig model

    DEFF Research Database (Denmark)

    Clausen, Morten Rahr; Christensen, Kirstine Lykke; Hedemann, Mette Skou

    2011-01-01

    outbred pigs. Results The metabolic phenotype of cloned pigs (n = 5) was for the first time elucidated by nuclear magnetic resonance (NMR)-based metabolomic analysis of multiple bio-fluids including plasma, bile and urine. The metabolic phenotype of the cloned pigs was compared with normal outbred pigs (n...

  15. cDNA cloning and characterization of a mannose-binding lectin from ...

    Indian Academy of Sciences (India)

    Using RNA extracted from Zingiber officinale rhizomes and primers designed according to the conservative regions of monocot mannose-binding lectins, the full-length cDNA of Z. officinale agglutinin (ZOA) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of zoa was 746 bp and contained a ...

  16. Caffeine, phenol and protein contents of thirty-seven clones of ...

    African Journals Online (AJOL)

    The coefficient of variation for pulp phenol was high (35.3), thus indicating that, rapid response to selection for favourable phenol percentage might be feasible. The high level of phenol found in some clones may be valuable in breeding for resistance to some major diseases and insect pests of coffee. There were differences ...

  17. [Human cloning in Muslim and Arab law].

    Science.gov (United States)

    Aldeeb Abu-Sahlieh, Sami A

    2009-01-01

    Cloning is a modern medical procedure that Muslim religious authorities treat en resorting to the general principles established by classical Muslim law based on the Koran and the Sunnah of Muhhamad as the messenger of God. In this regard, human beings are not capable of deciding what is or what is not lawful without resorting to divine norms. Cloning clashes with several principles. Firstly, the principle of the respect for life in relation to surpernumeraries, but Muslim authors are not in unanimous agreement on the determination of the moment at which life begins. Secondly, is the respect of progeny: cloning could only take place between a married couple. But even if these two principles are respected, cloning poses two major problems: the diversity of species expounded by the Koran and the Sunnah and a lack of interest. Which explains the quasi-unanimous opposition of Muslim writings regarding cloning.

  18. "Goodbye Dolly?" The ethics of human cloning.

    Science.gov (United States)

    Harris, J

    1997-01-01

    The ethical implications of human clones have been much alluded to, but have seldom been examined with any rigour. This paper examines the possible uses and abuses of human cloning and draws out the principal ethical dimensions, both of what might be done and its meaning. The paper examines some of the major public and official responses to cloning by authorities such as President Clinton, the World Health Organisation, the European parliament, UNESCO, and others and reveals their inadequacies as foundations for a coherent public policy on human cloning. The paper ends by defending a conception of reproductive rights of "procreative autonomy" which shows human cloning to be not inconsistent with human rights and dignity. PMID:9451604

  19. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach.

    Directory of Open Access Journals (Sweden)

    Hannes M Beyer

    Full Text Available Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly, a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date.

  20. 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…

  1. 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…

  2. 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.

  3. La réponse à la stimulation des clones d\\'une même classe ...

    African Journals Online (AJOL)

    Ceux à métabolisme lent s\\'expriment fortement, à la différence des clones à métabolisme rapide qui réagissent faiblement à cette même stimulation. Toutefois, il peut arriver que des clones d\\'une même classe métabolique se comportent différemment suite à la stimulation à l\\'Ethephon. La présente étude a pour objet ...

  4. 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.

  5. Identification of novel enzymes with different hydrolytic activities by metagenome expression cloning.

    Science.gov (United States)

    Lämmle, Katrin; Zipper, Hubert; Breuer, Michael; Hauer, Bernhard; Buta, Christiane; Brunner, Herwig; Rupp, Steffen

    2007-01-20

    Metagenome cloning has become a powerful tool to exploit the biocatalytic potential of microbial communities for the discovery of novel biocatalysts. In a novel variant of direct expression cloning, metagenomic DNA was isolated from compost by a modified direct lysis method, purified by size exclusion chromatography and cloned into an expression vector allowing bidirectional transcription. Transformation of Escherichia coli DH5alpha resulted in a metagenomic expression library with an average insert size of 3.2 kb. To estimate the functional diversity of the constructed library, it was screened by different approaches based on functional heterologous expression. A large number of active clones were identified, including lipolytic enzymes, amylases, phosphatases and dioxygenases. Molecular analysis of one important class of industrial biocatalysts, the lipolytic enzymes, confirmed the novelty and dissimilarity of all recovered genes, which exhibited only limited similarity to known enzymes. Equally, the novelty of another three genes encoding phosphatase or dioxygenase activity, respectively, was shown. These results demonstrate the suitability of this direct cloning approach, which comprised a dual-orientation expression vector and a simple one-step DNA purification method, for the efficient discovery of numerous active novel clones. By this means it provides an efficient way for the rapid generation of large libraries of hitherto unknown enzyme candidates which could be screened for different specific target reactions.

  6. Growth and nutrition of eucalyptus clones seedlings inoculated with mycorrhizal fungi

    Directory of Open Access Journals (Sweden)

    Francisco de Sousa Lima

    2014-06-01

    Full Text Available Eucalyptus is one of the most planted forest species, in Brazil, due to its rapid growth and high economic yield. Arbuscular mycorrhizal fungi improve the seedlings nutritional and phytosanitary status, besides increasing their resistance to biotic and abiotic stress. This study aimed to evaluate the effect of inoculation with arbuscular mycorrhizal fungi species on the growth and nutrition of different eucalyptus clones seedlings. The experiment was conducted under greenhouse conditions, in a randomized blocks design and a 5x5 factorial scheme (five fungal species and five eucalyptus clones, with five replications. In general, the mycorrhizal symbiosis significantly increased the growth and nutrition of eucalyptus seedlings, when compared to the non-inoculated seedlings. The most efficient interaction occured between the 2361 clone and the Entrophospora infrequens fungus, with increases of 107.3% and 120.6%, for the shoot and root dry biomass yield, and 107.7%, 94.1% and 103.3%, respectively for the accumulation of N, P and K in the seedlings shoots. All the fungal species studied showed a high absolute compatibility index with eucalyptus clones. The Glomus manihots and E. infrequens fungi presented a higher functional compatibility index with the clones tested. The 5204 clone showed 75% of compatibility with the fungi evaluated.

  7. Human embryo cloning prohibited in Hong Kong.

    Science.gov (United States)

    Liu, Athena

    2005-12-01

    Since the birth of Dolly (the cloned sheep) in 1997, debates have arisen on the ethical and legal questions of cloning-for-biomedical-research (more commonly termed "therapeutic cloning") and of reproductive cloning using human gametes. Hong Kong enacted the Human Reproductive Technology Ordinance (Cap 561) in 2000. Section 15(1)(e) of this Ordinance prohibits the "replacing of the nucleus of a cell of an embryo with a nucleus taken from any other cell," i.e., nucleus substitution. Section 15(1)(f) prohibits the cloning of any embryo. The scope of the latter, therefore, is arguably the widest, prohibiting all cloning techniques such as cell nucleus replacement, embryo splitting, parthenogenesis, and cloning using stem cell lines. Although the Human Reproductive Technology Ordinance is not yet fully operative, this article examines how these prohibitions may adversely impact on basic research and the vision of the Hong Kong scientific community. It concludes that in light of recent scientific developments, it is time to review if the law offers a coherent set of policies in this area.

  8. Serial cloning of pigs by somatic cell nuclear transfer: restoration of phenotypic normality during serial cloning.

    Science.gov (United States)

    Cho, Seong-Keun; Kim, Jae-Hwan; Park, Jong-Yi; Choi, Yun-Jung; Bang, Jae-Il; Hwang, Kyu-Chan; Cho, Eun-Jeong; Sohn, Sea-Hwan; Uhm, Sang Jun; Koo, Deog-Bon; Lee, Kyung-Kwang; Kim, Teoan; Kim, Jin-Hoi

    2007-12-01

    Somatic cell nuclear transfer (scNT) is a useful way to create cloned animals. However, scNT clones exhibit high levels of phenotypic instability. This instability may be due to epigenetic reprogramming and/or genomic damage in the donor cells. To test this, we produced transgenic pig fibroblasts harboring the truncated human thrombopoietin (hTPO) gene and used them as donor cells in scNT to produce first-generation (G1) cloned piglets. In this study, 2,818 scNT embryos were transferred to 11 recipients and five G1 piglets were obtained. Among them, a clone had a dimorphic facial appearance with severe hypertelorism and a broad prominent nasal bridge. The other clones looked normal. Second-generation (G2) scNT piglets were then produced using ear cells from a G1 piglet that had an abnormal nose phenotype. We reasoned that, if the phenotypic abnormality of the G1 clone was not present in the G2 and third-generation (G3) clones, or was absent in the G2 clones but reappeared in the G3 clones, the phenotypic instability of the G1 clone could be attributed to faulty epigenetic reprogramming rather than to inherent/accidental genomic damage to the donor cells. Blastocyst rates, cell numbers in blastocyst, pregnancy rates, term placenta weight and ponderal index, and birth weight between G1 and G2 clones did not differ, but were significantly (P clones and could not find any significant differences in the methylation patterns between G1 and G2 clones. Indeed, we failed to detect the phenotypic abnormality in the G2 and G3 clones. Thus, the phenotypic abnormality of the G1 clone is likely to be due to epigenetic dysregulation. Additional observations then suggested that expression of the hTPO gene in the transgenic clones did not appear to be the cause of the phenotypic abnormality in the G1 clones and that the abnormality was acquired by only a few of the G1 clone's cells during its gestational development. 2007 Wiley-Liss, Inc

  9. Altering the selection capabilities of common cloning vectors via restriction enzyme mediated gene disruption

    Science.gov (United States)

    2013-01-01

    Background The cloning of gene sequences forms the basis for many molecular biological studies. One important step in the cloning process is the isolation of bacterial transformants carrying vector DNA. This involves a vector-encoded selectable marker gene, which in most cases, confers resistance to an antibiotic. However, there are a number of circumstances in which a different selectable marker is required or may be preferable. Such situations can include restrictions to host strain choice, two phase cloning experiments and mutagenesis experiments, issues that result in additional unnecessary cloning steps, in which the DNA needs to be subcloned into a vector with a suitable selectable marker. Results We have used restriction enzyme mediated gene disruption to modify the selectable marker gene of a given vector by cloning a different selectable marker gene into the original marker present in that vector. Cloning a new selectable marker into a pre-existing marker was found to change the selection phenotype conferred by that vector, which we were able to demonstrate using multiple commonly used vectors and multiple resistance markers. This methodology was also successfully applied not only to cloning vectors, but also to expression vectors while keeping the expression characteristics of the vector unaltered. Conclusions Changing the selectable marker of a given vector has a number of advantages and applications. This rapid and efficient method could be used for co-expression of recombinant proteins, optimisation of two phase cloning procedures, as well as multiple genetic manipulations within the same host strain without the need to remove a pre-existing selectable marker in a previously genetically modified strain. PMID:23497512

  10. Evaluation of phenotypic stability of cassava clones by AMMI analysis in northwestern Paraná state

    Directory of Open Access Journals (Sweden)

    Marcus Vinícius Kvitschal

    2006-01-01

    Full Text Available High yield stability and adaptability of storage root are highly desirable attributes of cassava clones. Theobjective of this study was therefore to evaluate the effect of the genotype x environment interaction (G x E and the stability ofcassava clones developed at IAC. A subset of eight cassava genotypes was chosen in trials of storage root yield, arranged ina randomized complete block design with four replications, in two counties (Araruna and Maringá, in the northwesternregion of Paraná State, over five growing seasons (1997-2001. The G x E interaction was evaluated by joint varianceanalysis and stability and adaptability by AMMI analysis. The G x E interaction was significant (P<0.05 for storage rootyield. Results indicated AMMI analysis as an efficient tool for the evaluation of phenotypic adaptability and stability of cassavaclones and IAC 190 as the most promising clone.

  11. Variability of anatomical-physiological traits in black locust clones - Robinia pseudoacacia L

    Directory of Open Access Journals (Sweden)

    Orlović Saša S.

    2004-01-01

    Full Text Available Variability within R. pseudoacacia species represents an important factor in selection of fast-growing genotypes. Therefore, it is important to identify superior individuals according to their anatomical and physiological traits. This paper presents the results of a study of genotype variability of the main leaf anatomical (frequency, length and width of stomata, leaflet thickness among veins, leaflet thickness on the main vein, mesophyll thickness, length and width of vascular bundle of main vein and physiological (leaf area, photosynthetic pigments content and content of N P, K, Ca, Na parameters among five clones of Robinia pseudoacacia L. Significant interclonal variations were observed in the investigated parameters. Clone R-56 had the highest N, P, and K concentrations, the largest mesophyll volume and the highest pigment concentration. We concluded that the clone R-56, although without a remarkable leaf area, possesses the ability for high photosynthetic production. The results are going to be used in further work on selection.

  12. Simplified cryopreservation of porcine cloned blastocysts

    DEFF Research Database (Denmark)

    Du, Yutao; Zhang, Yunhai; Li, Juan

    2007-01-01

    )â€"handmade cloning (HMC)â€"to establish a simplified and efficient cryopreservation system for porcine cloned embryos. In Experiment 1, zonae pellucidae of oocytes were partially digested with pronase, followed by centrifugation to polarize lipid particles. Ninety percent (173/192) oocytes were successfully......). Our results prove that porcine embryos produced from delipated oocytes by PA or HMC can be cryopreserved effectively by ultrarapid vitrification. Further experiments are required to assess the in vivo developmental competence of the cloned-vitrified embryos  ...

  13. 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...

  14. 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.

  15. Growth and production of a short-rotation coppice culture of poplar - IV: Fine root characteristics of five poplar clones

    Energy Technology Data Exchange (ETDEWEB)

    Al Afas, N.; Marron, N.; Zavalloni, C.; Ceulemans, R. [Department of Biology, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk (Belgium)

    2008-06-15

    Below-ground characteristics of five Populus clones, belonging to different species and parentages, were studied during the second growing season of the third rotation of a high-density coppice culture. Size (length, area and volume), biomass, nitrogen and carbon concentrations of three classes of fine roots (diameter classes of 0-1, 1-2 and 2-5 mm) were determined for four different soil layers. Fine root biomass varied significantly among clones and among soil layers. Clone Primo (Populus deltoides x Populus nigra) had the highest root biomass and the longest fine roots, while clone Hazendans (Populus trichocarpa x P. deltoides) had the lowest root biomass and shortest fine roots. The topsoil layer (0-5 cm) was very rich in fine roots; the fine root biomass and distribution of all clones decreased with increasing soil depth. Fine root area index (diameter classes of 0-1 and 1-2 mm) varied among clones, with higher values for clones Wolterson and Primo (3.6 and 3.7, respectively), while clones Hazendans and Columbia River had lower values (1.7 and 2.2, respectively). The absence of a significant correlation between fine root traits and above-ground biomass leads to the conclusion that above-ground biomass was not a reliable indicator of below-ground biomass in poplar, probably because of the age of the plantation in our study (stump age of 10 years). Fine root area index was positively correlated with leaf area index for all clones and at all soil depths, i.e., clones with a high fine root area index also had a high leaf area index. We conclude that leaf area index can be an indicator of root area. (author)

  16. [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

  17. Cloning and characterization of the major histone H2A genes completes the cloning and sequencing of known histone genes of Tetrahymena thermophila.

    Science.gov (United States)

    Liu, X; Gorovsky, M A

    1996-01-01

    A truncated cDNA clone encoding Tetrahymena thermophila histone H2A2 was isolated using synthetic degenerate oligonucleotide probes derived from H2A protein sequences of Tetrahymena pyriformis. The cDNA clone was used as a homologous probe to isolate a truncated genomic clone encoding H2A1. The remaining regions of the genes for H2A1 (HTA1) and H2A2 (HTA2) were then isolated using inverse PCR on circularized genomic DNA fragments. These partial clones were assembled into intact HTA1 and HTA2 clones. Nucleotide sequences of the two genes were highly homologous within the coding region but not in the noncoding regions. Comparison of the deduced amino acid sequences with protein sequences of T. pyriformis H2As showed only two and three differences respectively, in a total of 137 amino acids for H2A1, and 132 amino acids for H2A2, indicating the two genes arose before the divergence of these two species. The HTA2 gene contains a TAA triplet within the coding region, encoding a glutamine residue. In contrast with the T. thermophila HHO and HTA3 genes, no introns were identified within the two genes. The 5'- and 3'-ends of the histone H2A mRNAs; were determined by RNase protection and by PCR mapping using RACE and RLM-RACE methods. Both genes encode polyadenylated mRNAs and are highly expressed in vegetatively growing cells but only weakly expressed in starved cultures. With the inclusion of these two genes, T. thermophila is the first organism whose entire complement of known core and linker histones, including replication-dependent and basal variants, has been cloned and sequenced. PMID:8760889

  18. Human cloning and the myth of disenchantment

    National Research Council Canada - National Science Library

    Laurențiu, Staicu

    2012-01-01

    This study has a twofold objective: firstly, it aims to examine the main types of argument that have been formulated against human cloning, to identify their presuppositions and to evaluate their strength...

  19. Cloning and molecular characterization of glyceraldehyde-3 ...

    African Journals Online (AJOL)

    Cloning and molecular characterization of glyceraldehyde-3-phosphate dehydrogenase gene from thermotolerant mushroom, Lentinus polychrous. ... sequence of the L. polychrous GAPDH-encoded protein shared significant sequence identity with other GAPDH proteins from basidiomycetes. Phylogenetic analysis ...

  20. DNA microarrays : a molecular cloning manual

    National Research Council Canada - National Science Library

    Sambrook, Joseph; Bowtell, David

    2002-01-01

    .... This manual, designed to extend and to complement the information in the best-selling Molecular Cloning, is a synthesis of the expertise and experience of more than 30 contributors all innovators in a fast moving field...

  1. Cloning and characterization of new bioluminescent proteins

    Science.gov (United States)

    Szent-Gyorgyi, Christopher; Ballou, Byron T.; Dagnal, Erich; Bryan, Bruce

    1999-07-01

    Over the past two years Prolume has undertaken a comprehensive program to clone luciferases and associated 'green fluorescent proteins' (GFPs) from marine animals that use coelenterazine as the luciferin. To data we have cloned several bioluminescent proteins, including two novel copepod luciferases and two anthozoan GFPs. These four proteins have sequences that differ greatly form previously cloned analogous proteins; the sequence diversity apparently is due to independent evolutionary origins and unusual evolutionary constraints. Thus coelenterazine-based bioluminescent systems may also manifest a variety of useful properties. We discuss form this taxonomic perspective the initial biochemical and spectral characterization of our cloned proteins. Emphasis is placed on the anthozoan luciferase-GFP systems, whose efficient resonance energy transfer has elicited much current interest.

  2. Meat and Milk Compositions of Bovine Clones

    National Research Council Canada - National Science Library

    X. Cindy Tian; Chikara Kubota; Kunihito Sakashita; Yoshiaki Izaike; Ryoichi Okano; Norio Tabara; Carol Curchoe; Lavina Jacob; Yuqin Zhang; Sadie Smith; Charles Bormann; Jie Xu; Masumi Sato; Sheila Andrew; Xiangzhong Yang; R. Michael Roberts

    2005-01-01

    ...? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction...

  3. Endangered wolves cloned from adult somatic cells.

    Science.gov (United States)

    Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Yuda, Fibrianto; Kim, Hye Jin; Hwang, Woo Suk; Hossein, Mohammad Shamim; Kim, Joung Joo; Shin, Nam Shik; Kang, Sung Keun; Lee, Byeong Chun

    2007-01-01

    Over the world, canine species, including the gray wolf, have been gradually endangered or extinct. Many efforts have been made to recover and conserve these canids. The aim of this study was to produce the endangered gray wolf with somatic cell nuclear transfer (SCNT) for conservation. Adult ear fibroblasts from a female gray wolf (Canis lupus) were isolated and cultured in vitro as donor cells. Because of limitations in obtaining gray wolf matured oocytes, in vivo matured canine oocytes obtained by flushing the oviducts from the isthmus to the infundibulum were used. After removing the cumulus cells, the oocyte was enucleated, microinjected, fused with a donor cell, and activated. The reconstructed cloned wolf embryos were transferred into the oviducts of the naturally synchronized surrogate mothers. Two pregnancies were detected by ultrasonography at 23 days of gestation in recipient dogs. In each surrogate dog, two fetal sacs were confirmed by early pregnancy diagnosis at 23 days, but only two cloned wolves were delivered. The first cloned wolf was delivered by cesarean section on October 18, 2005, 60 days after embryo transfer. The second cloned wolf was delivered on October 26, 2005, at 61 days postembryo transfer. Microsatellite analysis was performed with genomic DNA from the donor wolf, the two cloned wolves, and the two surrogate female recipients to confirm the genetic identity of the cloned wolves. Analysis of 19 microsatellite loci confirmed that the cloned wolves were genetically identical to the donor wolf. In conclusion, we demonstrated live birth of two cloned gray wolves by nuclear transfer of wolf somatic cells into enucleated canine oocyte, indicating that SCNT is a practical approach for conserving endangered canids.

  4. Variability in Growth, Physiological, and Biochemical Characteristics among Various Clones of Dalbergia sissoo in a Clonal Seed Orchard

    Directory of Open Access Journals (Sweden)

    Arvind Sharma

    2014-01-01

    Full Text Available Growth and physiological variability among clones of Dalbergia sissoo growing in a CSO revealed maximum height and GBH in Gonda clones (C196 and C198 and minimum growth attributes in Rajasthan clones. All biochemical constituents except sugar were also maximum in Gonda clones. Maximum chl. a, total chl., and chlorofluorescence (CF were recorded in C235 and C123 while chl. b was maximum in C198. Among tested clones, sugar content was maximum in C60 (Chhachhrauli while C198 (Gonda revealed maximum protein content. Heritability estimates of 8 characters at 99% revealed strong genetic control of total chls., sugars, proteins, and chl. b; however, maximum genetic gains of 117% and 80% were recorded for sugar and protein content, respectively. Correlation matrix revealed a positive correlation between height and GBH and CF. Among biochemical constituents, chl. a, and chl. b, chl. b, and total chl. were correlated significantly at 0.1% level. Regarding contribution of different parameters to variability, height and GBH were the greatest contributors. Clustering of clones on the basis of all three parameters separated clones in one major and six minor clusters. Average distance from centroid was found to be 22.61 whereas the maximum distance from centroid was 50.75.

  5. Diversity of leaf traits related to productivity in 31 Populus deltoides x Populus nigra clones.

    Science.gov (United States)

    Marron, Nicolas; Villar, Marc; Dreyer, Erwin; Delay, Didier; Boudouresque, Eric; Petit, Jean-Michel; Delmotte, Francis M; Guehl, Jean-Marc; Brignolas, Franck

    2005-04-01

    To test if some leaf parameters are predictors of productivity in a range of Populus deltoides (Bartr.) Marsh. x P. nigra L. clones, we assessed leaf traits and productivity in 2-month-old rooted cuttings from 31 clones growing in 4-l pots in a greenhouse, under conditions of controlled temperature and optimal irrigation. We evaluated four groups of variables describing (1) productivity (total biomass), (2) leaf growth (total leaf number increment and total leaf area increment rate), (3) leaf structure (specific leaf area and nitrogen and carbon contents) and (4) carbon isotope discrimination (delta), which is negatively correlated with time-integrated water-use efficiency. High-yielding clones did not necessarily display high leaf growth rates, but they displayed a larger total leaf area, lower specific leaf area and lower leaf nitrogen concentration than clones with low productivity. Total leaf area was mainly controlled by maximal individual leaf area and total leaf area increment rate (r = 0.51 and 0.56, respectively). Carbon isotope discrimination did not correlate with total biomass, but it was associated with total number of leaves and total leaf area increment rate (r = 0.39 and 0.45, respectively). Therefore, leaf area and specific leaf area were better indicators of productivity than leaf growth traits. The observed independence of delta from biomass production provides opportunities for selecting poplar clones combining high productivity and high water-use efficiency.

  6. Growth dynamics of rubber tree clones in northwestern Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Maria Isabela da Costa Terra

    2016-06-01

    Full Text Available ABSTRACT: This study aimed to evaluate the growth of rubber tree clones ( Hevea brasiliensis (Willd.exA.Juss. Müll. Arg. in the Northwest region of Minas Gerais, Brazil. Randomized blocks, constituted of three treatments (rubber tree clones and four repetitions were used, and evaluated as plots subdivided in time. Subplots represent the evaluations carried out at the ages 62, 77, 87, 99, 113 and 128 months after planting. It was evaluated survival, stem circumference and total plant height. Current annual increment and average annual increment of circumference and also the number of trees fit for rubber tapping were calculated. Results obtained indicate that clones GT 1, IAN 2880, IAC 15, RRIM 600, PB 235 and IAN 3156 have potential to establish under local edaphoclimatic conditions. Increase in plant height and stem circumference throughout the evaluation period makes it evident that the clones have different growing paces in the studied region. Clones GT 1, IAN 3087, IAN 3156 and PR 255 had larger stalk circumference at 128 months, and PR 255 was the only one fit for rubber tapping at the age of 128 months after planting.

  7. Emotional reactions to human reproductive cloning.

    Science.gov (United States)

    May, Joshua

    2016-01-01

    Extant surveys of people's attitudes towards human reproductive cloning focus on moral judgements alone, not emotional reactions or sentiments. This is especially important given that some (especially Leon Kass) have argued against such cloning on the ground that it engenders widespread negative emotions, like disgust, that provide a moral guide. To provide some data on emotional reactions to human cloning, with a focus on repugnance, given its prominence in the literature. This brief mixed-method study measures the self-reported attitudes and emotions (positive or negative) towards cloning from a sample of participants in the USA. Most participants condemned cloning as immoral and said it should be illegal. The most commonly reported positive sentiment was by far interest/curiosity. Negative emotions were much more varied, but anxiety was the most common. Only about a third of participants selected disgust or repugnance as something they felt, and an even smaller portion had this emotion come to mind prior to seeing a list of options. Participants felt primarily interested and anxious about human reproductive cloning. They did not primarily feel disgust or repugnance. This provides initial empirical evidence that such a reaction is not appropriately widespread. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  8. Reproductive cloning combined with genetic modification.

    Science.gov (United States)

    Strong, C

    2005-11-01

    Although there is widespread opposition to reproductive cloning, some have argued that its use by infertile couples to have genetically related children would be ethically justifiable. Others have suggested that lesbian or gay couples might wish to use cloning to have genetically related children. Most of the main objections to human reproductive cloning are based on the child's lack of unique nuclear DNA. In the future, it may be possible safely to create children using cloning combined with genetic modifications, so that they have unique nuclear DNA. The genetic modifications could be aimed at giving such children genetic characteristics of both members of the couple concerned. Thus, cloning combined with genetic modification could be appealing to infertile, lesbian, or gay couples who seek genetically related children who have genetic characteristics of both members. In such scenarios, the various objections to human reproductive cloning that are based on the lack of genetic uniqueness would no longer be applicable. The author argues that it would be ethically justifiable for such couples to create children in this manner, assuming these techniques could be used safely.

  9. CloneAssistant 1.0: a stand-alone software for automated cloning primer design.

    Science.gov (United States)

    Shao, Chaogang; Meng, Yijun; Lv, Shaolei; Zhong, Wei; Wang, Zheyu; Chen, Ming

    2010-11-01

    "CloneAssistant 1.0" is a stand-alone software compatible with the current Windows operating systems, which can automatically design cloning primers with full consideration of the sequence information of vectors and genes, cloning strategies, the principles of primer design, reading frames, position effects, and enzymatic reaction conditions for users. Five internal XML (extensible markup language) databases [restriction enzymes, plasmids, universal buffers, PCR (polymerase chain reaction) protection bases, and an MCS (multiple cloning site) double digest interference database] were established to serve as the basic support for "CloneAssistant 1.0". The primer pairs designed are sorted according to the difficulty of the follow-up experiments. Once a primer pair is selected by the user, detailed experimental guidance for this primer pair will be provided. In addition, "CloneAssistant 1.0" can be used for restriction map analysis, ORF (open reading frame) finding, sequence alignment and complementary analysis, translation, restriction enzyme and universal buffer queries, and isocaudamer analysis. "CloneAssistant 1.0" makes gene clone design much easier, and it can be freely downloaded from http://bis.zju.edu.cn/clone. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Birth of viable puppies derived from breeding cloned female dogs with a cloned male.

    Science.gov (United States)

    Park, J E; Hong, S G; Kang, J T; Oh, H J; Kim, M K; Kim, M J; Kim, H J; Kim, D Y; Jang, G; Lee, B C

    2009-09-15

    Since the establishment of production of viable cloned dogs by somatic cell nucleus transfer, great concern has been given to the reproductive abilities of these animals (Canis familiaris). Therefore, we investigated reproductive activity of cloned dogs by (1) performing sperm analysis using computer-assisted sperm analysis and early embryonic development, (2) assessing reproductive cycling by measuring serum progesterone (P4) levels and performing vaginal cytology, and (3) breeding cloned dogs using artificial insemination. Results showed that most parameters of sperm motility in a cloned male dog were within the reference range, and in vivo-matured oocytes from a noncloned female were successfully fertilized by spermatozoa from a cloned male dog and develop normally to the 8-cell stage. Three cloned female dogs displayed normal patterns of P4 levels and morphologic changes of the vaginal epithelium. Two cloned female dogs became pregnant using semen from a cloned male dog and successfully delivered 10 puppies by natural labor. In conclusion, these data demonstrated that both cloned male and female dogs are fertile, and their puppies are currently alive and healthy with normal growth patterns.

  11. Do Clones Dream of Love? Images of Clones in Popular Culture

    Directory of Open Access Journals (Sweden)

    Dragana Antonijević

    2016-02-01

    Full Text Available Fantasies about clones, cyborgs and androids have become part and parcel of the mythology of modern times – the mythologies of the biotechnological era in which the achievements of genetic engineering have inflamed fears of possible abuse of scientific knowledge and the consequences of such abuse. The paper considers the phenomenon of reproductive cloning of human beings as it is represented in popular culture, especially film as it is one of the most important sources of representations and constructions of ideas about clones. After the introductory consideration of this phenomenon in scientific, ethical and media debates which are imbued with rejection of reproductive cloning, I have analyzed the different uses of the clone motif in selected movies. I have examined the structure and content of the genre formula of "social melodrama" which is present in films about clones, and have analyzed the mythical patterns pertaining to the topic of cloning, such as the myth of immortality, the myth of twins, the myth of the uniqueness of human kind etc. Ultimately, the nature and origins of the fear of clones and disgust that clones cause have been examined, and it has been shown that they mostly boil down to the fear of the dehumanization of human beings, the fear of the loss of difference and the transgression of biological, sociocultural and metaphysical boundaries.

  12. Cloning higher plants from aseptically cultured tissues and cells

    Science.gov (United States)

    Krikorian, A. D.

    1982-01-01

    A review of aseptic culture methods for higher plants is presented, which focuses on the existing problems that limit or prevent the full realization of cloning plants from free cells. It is shown that substantial progress in clonal multiplication has been made with explanted stem tips or lateral buds which can be stimulated to produce numerous precocious axillary branches. These branches can then be separated or subdivided and induced to root in order to yield populations of genetically and phenotypically uniorm plantlets. Similarly, undifferentiated calluses can sometimes be induced to form shoots and/or roots adventitiously. Although the cell culture techniques required to produce somatic embryos are presently rudimentary, steady advances are being made in learning how to stimulate formation of somatic or adventive embryos from totipotent cells grown in suspension cultures. It is concluded that many problems exist in the producing and growing of totipotent or morphogenetically competent cell suspensions, but the potential benefits are great.

  13. Popular Theatre for Science Engagement: Audience Engagement with Human Cloning Following a Production of Caryl Churchill's "A Number"

    Science.gov (United States)

    Donkers, Martina; Orthia, Lindy A.

    2016-01-01

    Research into the role of fiction in engaging people with science is a growing area, but a little studied medium in this respect is "popular theatre," or non-pedagogic theatre that exists primarily as a work of art. This study investigated audience engagement with human cloning issues after seeing a performance of Caryl Churchill's 2002…

  14. Public perceptions of farm animal cloning in Europe

    DEFF Research Database (Denmark)

    Lassen, Jesper

    This report presents a picture of European opinion on farm animal cloning. In the report, both agricultural and biomedical applications of farm animal cloning are considered. With the arrival of Dolly, animal cloning became an integral part of the biotech debate, but this debate did not isolate...... animal cloning as a single issue....

  15. Cloning: Past, Present, and the Exciting Future. Breakthroughs in Bioscience.

    Science.gov (United States)

    Di Berardino, Marie A.

    This document explores the history of cloning by focusing on Dolly the Sheep, one of the first large animal clonings. The disadvantages and advantages of transgenic clones are discussed as well as the future implications of cloning from the perspective of human health. (Contains 10 resources.) (YDS)

  16. AN APPROACH FOR CLONE DETECTION IN DOCUMENTATION REUSE

    Directory of Open Access Journals (Sweden)

    D. V. Lutsiv

    2014-07-01

    Full Text Available The paper focuses on the searching method for repetitions in DocBook/DRL or plain text documents. An algorithm has been designed based on software clone detection. The algorithm supports filtering results: clones are rejected if clone length in the group is less than 5 symbols, intersection of clone groups is eliminated, meaningfulness clones are removed, the groups containing clones consisting only of XML are eliminated. Remaining search is supported: found clones are extracted from the documentation, and clone search is repeated. One step is proved to be enough. Adaptive reuse technique of Paul Bassett – Stan Jarzabek has been implemented. A software tool has been developed on the basis of the algorithm. The tool supports setting parameters for repetitions detection and visualization of the obtained results. The tool is integrated into DocLine document development environment, and provides refactoring of documents using found clones. The Clone Miner clone detection utility is used for clones search. The method has been evaluated for Linux Kernel Documentation (29 documents, 25000 lines. Five semantic kinds of clones have been selected: terms (abbreviations, one word and two word terms, hyperlinks, license agreements, functionality description, and code examples. 451 meaningful clone groups have been found, average clone length is 4.43 tokens, and average number of clones in a group is 3.56.

  17. Biotechnology and genetic optimization of fast-growing hardwoods

    Energy Technology Data Exchange (ETDEWEB)

    Garton, S.; Syrkin-Wurtele, E.; Griffiths, H.; Schell, J.; Van Camp, L.; Bulka, K. (NPI, Salt Lake City, UT (United States))

    1991-02-01

    A biotechnology research program was initiated to develop new clones of fast-growing Populus clones resistant to the herbicide glyphosate and resistant to the leaf-spot and canker disease caused by the fungus Septoria musiva. Glyphosate-resistant callus was selected from stem segments cultured in vitro on media supplemented with the herbicide. Plants were regenerated from the glyphosate-resistant callus tissue. A portion of plants reverted to a glyphosate susceptible phenotype during organogenesis. A biologically active filtrate was prepared from S. musiva and influenced fresh weight of Populus callus tissue. Disease-resistant plants were produced through somaclonal variation when shoots developed on stem internodes cultured in vitro. Plantlets were screened for disease symptoms after spraying with a suspension of fungal spores. A frequency of 0.83 percent variant production was observed. Genetically engineered plants were produced after treatment of plant tissue with Agrobacterium tumefasciens strains carrying plasmid genes for antibiotic resistance. Transformers were selected on media enriched with the antibiotic, kanamycin. Presence of foreign DNA was confirmed by Southern blot analysis. Protoplasts of popular were produced but did not regenerate into plant organs. 145 refs., 12 figs., 36 tabs.

  18. Drought tolerance is associated with rooting depth and stomatal control of water use in clones of Coffea canephora.

    Science.gov (United States)

    Pinheiro, Hugo A; Damatta, Fábio M; Chaves, Agnaldo R M; Loureiro, Marcelo E; Ducatti, Carlos

    2005-07-01

    Drought is a major environmental constraint affecting growth and production of Coffea canephora. Selection of C. canephora clones has been largely empirical as little is known about how clones respond physiologically to drought. Using clones previously shown to differ in drought tolerance, this study aimed to identify the extent of variation of water use and the mechanisms responsible, particularly those associated morphological traits. * Clones (14 and 120, drought-tolerant; 46 and 109A, drought-sensitive, based on their abilities to yield under drought) were grown in 120-L pots until they were 12-months old, when an irrigation and a drought treatment were applied; plants were droughted until the pressure potential (psi(x)) before dawn (pre-dawn) reached -3.0 MPa. Throughout the drought period, psi(x) and stomatal conductance (g(s)) were measured. At the end of the experiment, carbon isotope ratio and parameters from pressure-volume curves were estimated. Morphological traits were also assessed. * With irrigation, plant hydraulic conductance (K(L)), midday psi(x) and total biomass were all greater in clones 109A and 120 than in the other clones. Root mass to leaf area ratio was larger in clone 109A than in the others, whereas rooting depth was greater in drought-tolerant than in drought-sensitive clones. Predawn psi(x) of -3.0 MPa was reached fastest by 109A, followed progressively by clones 46, 120 and 14. Decreases in g(s) with declining psi(x), or increasing evaporative demand, were similar for clones 14, 46, and 120, but lower in 109A. Carbon isotope ratio increased under drought; however, it was lower in 109A than in other clones. For all clones, psi(x), g(s) and K(L) recovered rapidly following re-watering. Differences in root depth, K(L) and stomatal control of water use, but not osmotic or elastic adjustments, largely explained the differences in relative tolerance to drought stress of clones 14 and 120 compared with clones 46 and 109A.

  19. Development and Characterization of Recombinant Virus Generated from a New World Zika Virus Infectious Clone.

    Science.gov (United States)

    Weger-Lucarelli, James; Duggal, Nisha K; Bullard-Feibelman, Kristen; Veselinovic, Milena; Romo, Hannah; Nguyen, Chilinh; Rückert, Claudia; Brault, Aaron C; Bowen, Richard A; Stenglein, Mark; Geiss, Brian J; Ebel, Gregory D

    2017-01-01

    Zika virus (ZIKV; family Flaviviridae, genus Flavivirus) is a rapidly expanding global pathogen that has been associated with severe clinical manifestations, including devastating neurological disease in infants. There are currently no molecular clones of a New World ZIKV available that lack significant attenuation, hindering progress toward understanding determinants of transmission and pathogenesis. Here we report the development and characterization of a novel ZIKV reverse genetics system based on a 2015 isolate from Puerto Rico (PRVABC59). We generated a two-plasmid infectious clone system from which infectious virus was rescued that replicates in human and mosquito cells with growth kinetics representative of wild-type ZIKV. Infectious clone-derived virus initiated infection and transmission rates in Aedes aegypti mosquitoes comparable to those of the primary isolate and displayed similar pathogenesis in AG129 mice. This infectious clone system provides a valuable resource to the research community to explore ZIKV molecular biology, vaccine development, antiviral development, diagnostics, vector competence, and disease pathogenesis. ZIKV is a rapidly spreading mosquito-borne pathogen that has been linked to Guillain-Barré syndrome in adults and congenital microcephaly in developing fetuses and infants. ZIKV can also be sexually transmitted. The viral molecular determinants of any of these phenotypes are not well understood. There is no reverse genetics system available for the current epidemic virus that will allow researchers to study ZIKV immunity, develop novel vaccines, or develop antiviral drugs. Here we provide a novel infectious clone system generated from a recent ZIKV isolated from a patient infected in Puerto Rico. This infectious clone produces virus with in vitro and in vivo characteristics similar to those of the primary isolate, providing a critical tool to study ZIKV infection and disease. Copyright © 2016 American Society for Microbiology.

  20. Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system.

    Science.gov (United States)

    Krebber, A; Bornhauser, S; Burmester, J; Honegger, A; Willuda, J; Bosshard, H R; Plückthun, A

    1997-02-14

    A prerequisite for the use of recombinant antibody technologies starting from hybridomas or immune repertoires is the reliable cloning of functional immunoglobulin genes. For this purpose, a standard phage display system was optimized for robustness, vector stability, tight control of scFv-delta geneIII expression, primer usage for PCR amplification of variable region genes, scFv assembly strategy and subsequent directional cloning using a single rare cutting restriction enzyme. This integrated cloning, screening and selection system allowed us to rapidly obtain antigen binding scFvs derived from spleen-cell repertoires of mice immunized with ampicillin as well as from all hybridoma cell lines tested to date. As representative examples, cloning of monoclonal antibodies against a his tag, leucine zippers, the tumor marker EGP-2 and the insecticide DDT is presented. Several hybridomas whose genes could not be cloned in previous experimental setups, but were successfully obtained with the present system, expressed high amounts of aberrant heavy and light chain mRNAs, which were amplified by PCR and greatly exceeded the amount of binding antibody sequences. These contaminating variable region genes were successfully eliminated by employing the optimized phage display system, thus avoiding time consuming sequencing of non-binding scFv genes. To maximize soluble expression of functional scFvs subsequent to cloning, a compatible vector series to simplify modification, detection, multimerization and rapid purification of recombinant antibody fragments was constructed.

  1. 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.

  2. Phenomenon of Cloning and specificity of its usage

    OpenAIRE

    Yusubova, Zyliya; Zeremskaya, Yuliya Aleksandrovna

    2017-01-01

    Cloning is studied by different branches of science. Medicine is interested in cloning because of its ability to transplant special tissues and organs, genetics - with the purpose of studying heredity and succession, sociology deals with moral and ethic aspects of the phenomenon. The paper is devoted to the study of cloning, its special features and usage in different spheres of social life. The article represents main types of cloning, specificity of vegetative and animal cloning and problem...

  3. 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...

  4. 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.

  5. 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...

  6. 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.

  7. 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.

  8. Cloning mice and men: prohibiting the use of iPS cells for human reproductive cloning.

    Science.gov (United States)

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2010-01-08

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation. Copyright 2010 Elsevier Inc. All rights reserved.

  9. Update on the First Cloned Dog and Outlook for Canine Cloning.

    Science.gov (United States)

    Jang, Goo; Lee, ByeongChun

    2015-10-01

    As man's best friend, dogs have an important position in human society. Ten years ago, we reported the first cloned dog, and his birth has raised various scientific issues, such as those related to health, reproduction, and life span. He has developed without any unique health issues. In this article, we summarize and present perspectives on canine cloning.

  10. Variations and voids: the regulation of human cloning around the world.

    Science.gov (United States)

    Pattinson, Shaun D; Caulfield, Timothy

    2004-12-13

    No two countries have adopted identical regulatory measures on cloning. Understanding the complexity of these regulatory variations is essential. It highlights the challenges associated with the regulation of a controversial and rapidly evolving area of science and sheds light on a regulatory framework that can accommodate this reality. Using the most reliable information available, we have performed a survey of the regulatory position of thirty countries around the world regarding the creation and use of cloned embryos (see Table 1). We have relied on original and translated legislation, as well as published sources and personal communications. We have examined the regulation of both reproductive cloning (RC) and non-reproductive cloning (NRC). While most of the countries studied have enacted national legislation, the absence of legislation in seven of these countries should not be equated with the absence of regulation. Senator Morin was not correct in stating that the majority of recent legislation bans both RC and NRC. Recent regulatory moves are united only with regard to the banning of RC. While NRC is not permitted in seventeen of the countries examined, it could be permitted in up to thirteen countries. There is little consensus on the various approaches to cloning laws and policies, and the regulatory position in many countries remains uncertain.

  11. Optical Barcoding for Single-Clone Tracking to Study Tumor Heterogeneity.

    Science.gov (United States)

    Mohme, Malte; Maire, Cecile L; Riecken, Kristoffer; Zapf, Svenja; Aranyossy, Tim; Westphal, Manfred; Lamszus, Katrin; Fehse, Boris

    2017-03-01

    Intratumoral heterogeneity has been identified as one of the strongest drivers of treatment resistance and tumor recurrence. Therefore, investigating the complex clonal architecture of tumors over time has become a major challenge in cancer research. We developed a new fluorescent "optical barcoding" technique that allows fast tracking, identification, and quantification of live cell clones in vitro and in vivo using flow cytometry (FC). We optically barcoded two cell lines derived from malignant glioma, an exemplary heterogeneous brain tumor. In agreement with mathematical combinatorics, we demonstrate that up to 41 clones can unambiguously be marked using six fluorescent proteins and a maximum of three colors per clone. We show that optical barcoding facilitates sensitive, precise, rapid, and inexpensive analysis of clonal composition kinetics of heterogeneous cell populations by FC. We further assessed the quantitative contribution of multiple clones to glioblastoma growth in vivo and we highlight the potential to recover individual viable cell clones by fluorescence-activated cell sorting. In summary, we demonstrate that optical barcoding is a powerful technique for clonal cell tracking in vitro and in vivo, rendering this approach a potent tool for studying the heterogeneity of complex tissues, in particular, cancer. Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

  12. Efficient four fragment cloning for the construction of vectors for targeted gene replacement in filamentous fungi

    Directory of Open Access Journals (Sweden)

    Kristensen Matilde B

    2008-08-01

    Full Text Available Abstract Background The rapid increase in whole genome fungal sequence information allows large scale functional analyses of target genes. Efficient transformation methods to obtain site-directed gene replacement, targeted over-expression by promoter replacement, in-frame epitope tagging or fusion of coding sequences with fluorescent markers such as GFP are essential for this process. Construction of vectors for these experiments depends on the directional cloning of two homologous recombination sequences on each side of a selection marker gene. Results Here, we present a USER Friendly cloning based technique that allows single step cloning of the two required homologous recombination sequences into different sites of a recipient vector. The advantages are: A simple experimental design, free choice of target sequence, few procedures and user convenience. The vectors are intented for Agrobacterium tumefaciens and protoplast based transformation technologies. The system has been tested by the construction of vectors for targeted replacement of 17 genes and overexpression of 12 genes in Fusarium graminearum. The results show that four fragment vectors can be constructed in a single cloning step with an average efficiency of 84% for gene replacement and 80% for targeted overexpression. Conclusion The new vectors designed for USER Friendly cloning provided a fast reliable method to construct vectors for targeted gene manipulations in fungi.

  13. A one pot, one step, precision cloning method with high throughput capability.

    Directory of Open Access Journals (Sweden)

    Carola Engler

    Full Text Available Current cloning technologies based on site-specific recombination are efficient, simple to use, and flexible, but have the drawback of leaving recombination site sequences in the final construct, adding an extra 8 to 13 amino acids to the expressed protein. We have devised a simple and rapid subcloning strategy to transfer any DNA fragment of interest from an entry clone into an expression vector, without this shortcoming. The strategy is based on the use of type IIs restriction enzymes, which cut outside of their recognition sequence. With proper design of the cleavage sites, two fragments cut by type IIs restriction enzymes can be ligated into a product lacking the original restriction site. Based on this property, a cloning strategy called 'Golden Gate' cloning was devised that allows to obtain in one tube and one step close to one hundred percent correct recombinant plasmids after just a 5 minute restriction-ligation. This method is therefore as efficient as currently used recombination-based cloning technologies but yields recombinant plasmids that do not contain unwanted sequences in the final construct, thus providing precision for this fundamental process of genetic manipulation.

  14. GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.

    Directory of Open Access Journals (Sweden)

    Athanasios Lampropoulos

    Full Text Available Building expression constructs for transgenesis is one of the fundamental day-to-day tasks in modern biology. Traditionally it is based on a multitude of type II restriction endonucleases and T4 DNA ligase. Especially in case of long inserts and applications requiring high-throughput, this approach is limited by the number of available unique restriction sites and the need for designing individual cloning strategies for each project. Several alternative cloning systems have been developed in recent years to overcome these issues, including the type IIS enzyme based Golden Gate technique. Here we introduce our GreenGate system for rapidly assembling plant transformation constructs, which is based on the Golden Gate method. GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules (plant promoter, N-terminal tag, coding sequence, C-terminal tag, plant terminator and plant resistance cassette, but at the same time allows assembling several expression cassettes in one binary destination vector from a collection of pre-cloned building blocks. The system is cheap and reliable and when combined with a library of modules considerably speeds up cloning and transgene stacking for plant transformation.

  15. GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.

    Science.gov (United States)

    Lampropoulos, Athanasios; Sutikovic, Zoran; Wenzl, Christian; Maegele, Ira; Lohmann, Jan U; Forner, Joachim

    2013-01-01

    Building expression constructs for transgenesis is one of the fundamental day-to-day tasks in modern biology. Traditionally it is based on a multitude of type II restriction endonucleases and T4 DNA ligase. Especially in case of long inserts and applications requiring high-throughput, this approach is limited by the number of available unique restriction sites and the need for designing individual cloning strategies for each project. Several alternative cloning systems have been developed in recent years to overcome these issues, including the type IIS enzyme based Golden Gate technique. Here we introduce our GreenGate system for rapidly assembling plant transformation constructs, which is based on the Golden Gate method. GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules (plant promoter, N-terminal tag, coding sequence, C-terminal tag, plant terminator and plant resistance cassette), but at the same time allows assembling several expression cassettes in one binary destination vector from a collection of pre-cloned building blocks. The system is cheap and reliable and when combined with a library of modules considerably speeds up cloning and transgene stacking for plant transformation.

  16. Functional annotation of 19,841 Populus nigra full-length enriched cDNA clones

    Science.gov (United States)

    Nanjo, Tokihiko; Sakurai, Tetsuya; Totoki, Yasushi; Toyoda, Atsushi; Nishiguchi, Mitsuru; Kado, Tomoyuki; Igasaki, Tomohiro; Futamura, Norihiro; Seki, Motoaki; Sakaki, Yoshiyuki; Shinozaki, Kazuo; Shinohara, Kenji

    2007-01-01

    Background Populus is one of favorable model plants because of its small genome. Structural genomics of Populus has reached a breakpoint as nucleotides of the entire genome have been determined. Reaching the post genome era, functional genomics of Populus is getting more important for well-comprehended plant science. Development of bioresorce serving functional genomics is making rapid progress. Huge efforts have achieved deposits of expressed sequence tags (ESTs) in various plant species consequently accelerating functional analysis of genes. ESTs from full-length cDNA clones are especially powerful for accurate molecular annotation. We promoted collection and annotation of the ESTs from Populus full-length enriched cDNA clones as part of functional genomics of tree species. Results We have been collecting the full-length enriched cDNA of the female poplar (Populus nigra var. italica) for years. By sequencing P. nigra full-length (PnFL) cDNA libraries, we generated about 116,000 5'-end or 3'-end ESTs corresponding to 19,841 nonredundant PnFL clones. Population of PnFL cDNA clones represents 44% of the predicted genes in the Populus genome. Conclusion Our resource of P. nigra full-length enriched clones is expected to provide valuable tools to gain further insight into genome annotation and functional genomics in Populus. PMID:18053163

  17. Functional annotation of 19,841 Populus nigra full-length enriched cDNA clones

    Directory of Open Access Journals (Sweden)

    Seki Motoaki

    2007-12-01

    Full Text Available Abstract Background Populus is one of favorable model plants because of its small genome. Structural genomics of Populus has reached a breakpoint as nucleotides of the entire genome have been determined. Reaching the post genome era, functional genomics of Populus is getting more important for well-comprehended plant science. Development of bioresorce serving functional genomics is making rapid progress. Huge efforts have achieved deposits of expressed sequence tags (ESTs in various plant species consequently accelerating functional analysis of genes. ESTs from full-length cDNA clones are especially powerful for accurate molecular annotation. We promoted collection and annotation of the ESTs from Populus full-length enriched cDNA clones as part of functional genomics of tree species. Results We have been collecting the full-length enriched cDNA of the female poplar (Populus nigra var. italica for years. By sequencing P. nigra full-length (PnFL cDNA libraries, we generated about 116,000 5'-end or 3'-end ESTs corresponding to 19,841 nonredundant PnFL clones. Population of PnFL cDNA clones represents 44% of the predicted genes in the Populus genome. Conclusion Our resource of P. nigra full-length enriched clones is expected to provide valuable tools to gain further insight into genome annotation and functional genomics in Populus.

  18. Pathogenicity of a Very Virulent Strain of Marek's Disease Herpesvirus Cloned as Infectious Bacterial Artificial Chromosomes

    Directory of Open Access Journals (Sweden)

    Lorraine P. Smith

    2011-01-01

    Full Text Available Bacterial artificial chromosome (BAC vectors containing the full-length genomes of several herpesviruses have been used widely as tools to enable functional studies of viral genes. Marek's disease viruses (MDVs are highly oncogenic alphaherpesviruses that induce rapid-onset T-cell lymphomas in chickens. Oncogenic strains of MDV reconstituted from BAC clones have been used to examine the role of viral genes in inducing tumours. Past studies have demonstrated continuous increase in virulence of MDV strains. We have previously reported on the UK isolate C12/130 that showed increased virulence features including lymphoid organ atrophy and enhanced tropism for the central nervous system. Here we report the construction of the BAC clones (pC12/130 of this strain. Chickens were infected with viruses reconstituted from the pC12/130 clones along with the wild-type virus for the comparison of the pathogenic properties. Our studies show that BAC-derived viruses induced disease similar to the wild-type virus, though there were differences in the levels of pathogenicity between individual viruses. Generation of BAC clones that differ in the potential to induce cytolytic disease provide the opportunity to identify the molecular determinants of increased virulence by direct sequence analysis as well as by using reverse genetics approaches on the infectious BAC clones.

  19. Variations and voids: the regulation of human cloning around the world

    Directory of Open Access Journals (Sweden)

    Caulfield Timothy

    2004-12-01

    Full Text Available Abstract Background No two countries have adopted identical regulatory measures on cloning. Understanding the complexity of these regulatory variations is essential. It highlights the challenges associated with the regulation of a controversial and rapidly evolving area of science and sheds light on a regulatory framework that can accommodate this reality. Methods Using the most reliable information available, we have performed a survey of the regulatory position of thirty countries around the world regarding the creation and use of cloned embryos (see Table 1. We have relied on original and translated legislation, as well as published sources and personal communications. We have examined the regulation of both reproductive cloning (RC and non-reproductive cloning (NRC. Results While most of the countries studied have enacted national legislation, the absence of legislation in seven of these countries should not be equated with the absence of regulation. Senator Morin was not correct in stating that the majority of recent legislation bans both RC and NRC. Recent regulatory moves are united only with regard to the banning of RC. While NRC is not permitted in seventeen of the countries examined, it could be permitted in up to thirteen countries. Conclusions There is little consensus on the various approaches to cloning laws and policies, and the regulatory position in many countries remains uncertain.

  20. Benefits and problems with cloning animals.

    Science.gov (United States)

    Smith, L C; Bordignon, V; Babkine, M; Fecteau, G; Keefer, C

    2000-01-01

    Animal cloning is becoming a useful technique for producing transgenic farm animals and is likely to be used to produce clones from valuable adults. Other applications will also undoubtedly be discovered in the near future, such as for preserving endangered breeds and species. Although cloning promises great advantages for commerce and research alike, its outcome is not always certain due to high pregnancy losses and high morbidity and mortality during the neonatal period. Research into the mechanisms involved in the reprogramming of the nucleus is being conducted throughout the world in an attempt to better understand the molecular and cellular mechanisms involved in correcting these problems. Although the cause of these anomalies remains mostly unknown, similar phenotypes have been observed in calves derived through in vitro fertilization, suggesting that culture conditions are involved in these phenomena. In the meantime, veterinarians and theriogenologists have an important role to play in improving the efficiency of cloning by finding treatments to assure normal gestation to term and to develop preventative and curative care for cloned neonates. Images Figure 1. PMID:11143925

  1. Elephant grass clones for silage production

    Directory of Open Access Journals (Sweden)

    Rerisson José Cipriano dos Santos

    2013-02-01

    Full Text Available Ensiling warm-season grasses often requires wilting due to their high moisture content, and the presence of low-soluble sugars in these grasses usually demands the use of additives during the ensiling process. This study evaluated the bromatological composition of the fodder and silage from five Pennisetum sp. clones (IPA HV 241, IPA/UFRPE Taiwan A-146 2.114, IPA/UFRPE Taiwan A-146 2.37, Elephant B, and Mott. The contents of 20 Polyvinyl chloride (PVC silos, which were opened after 90 days of storage, were used for the bromatological analysis and the evaluation of the pH, nitrogen, ammonia, buffer capacity, soluble carbohydrates, and fermentation coefficients. The effluent losses, gases and dry matter recovery were also calculated. Although differences were observed among the clones (p < 0.05 for the concentrations of dry matter, insoluble nitrogen in acid detergents, insoluble nitrogen in neutral detergents, soluble carbohydrates, fermentation coefficients, and in vitro digestibility in the forage before ensiling, no differences were observed for most of these variables after ensiling. All of the clones were efficient in the fermentation process. The IPA/UFRPE TAIWAN A-146 2.37 clone, however, presented a higher dry matter concentration and the best fermentation coefficient, resulting in a better silage quality, compared to the other clones.

  2. 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,

  3. Cloning and Heterologous Expression of the Grecocycline Biosynthetic Gene Cluster.

    Directory of Open Access Journals (Sweden)

    Oksana Bilyk

    Full Text Available Transformation-associated recombination (TAR in yeast is a rapid and inexpensive method for cloning and assembly of large DNA fragments, which relies on natural homologous recombination. Two vectors, based on p15a and F-factor replicons that can be maintained in yeast, E. coli and streptomycetes have been constructed. These vectors have been successfully employed for assembly of the grecocycline biosynthetic gene cluster from Streptomyces sp. Acta 1362. Fragments of the cluster were obtained by PCR and transformed together with the "capture" vector into the yeast cells, yielding a construct carrying the entire gene cluster. The obtained construct was heterologously expressed in S. albus J1074, yielding several grecocycline congeners. Grecocyclines have unique structural moieties such as a dissacharide side chain, an additional amino sugar at the C-5 position and a thiol group. Enzymes from this pathway may be used for the derivatization of known active angucyclines in order to improve their desired biological properties.

  4. Beginnings of fruit growing in the old world.

    Science.gov (United States)

    Zohary, D; Spiegel-Roy, P

    1975-01-31

    The article reviews the available information on the start of fruit tree cultivation in the Old World. On the basis of (i) evaluation of the available archeological remains and (ii) examination of the wild relatives of the cultivated crops, it was concluded that olive, grape, date, and fig were the first important horticultural additions to the Mediterranean grain agriculture. They were most likely domesticated in the Near East in protohistoric time (fourth and third millennia B.C.) and they emerge as important food elements in the early Bronze Age. Domestication of all four fruit trees was based on a shift from sexual reproduction (in the wild) to vegetative propagation of clones (under domestication). Olive, grape, date, and fig can be vegetatively propagated by simple techniques (cuttings, basal knobs, suckers) and were thus preadapted for domestication early in the development of agriculture. The shift to clonal propagation placed serious limitations on selection and on fruit set under cultivation. We have examined the consequences of this shift in terms of the genetic makeup of the cultivars and traced the various countermeasures that evolved to ensure fruit set. Finally, it was pointed out that in each of these classic fruit trees we are confronted with a variable complex of genuinely wild types, secondary weedy derivatives and feral plants, and groups of the domesticated clones, which are all interfertile and interconnected by occasional hybridization. It was concluded that introgression from the diversified wild gene pool facilitated the rapid buildup of variation in the domesticated crops.

  5. Scientific hazards of human reproductive 'cloning'.

    Science.gov (United States)

    Young, Lorraine E

    2003-05-01

    The scientific and clinical professional societies and associations covering the remit of Human Fertility are unanimously opposed to human reproductive 'cloning'. This article describes the main scientific objections to human reproductive 'cloning'. Data collected from numerous studies in a range of animal species indicate a high incidence of fetal defects, a stillbirth rate typically of more than 90% and a lack of adequate information on postnatal development. These concerns are exacerbated by misconceptions about the current ability to screen preimplantation embryos for 'cloning-induced' defects. Scientists and clinicians are sometimes treated with mistrust in the eyes of the public and media over such issues, perhaps because scientific information is not as well communicated as it might be. The duty of reproductive specialists is to convey the limits of their knowledge on this issue to the public and policymakers.

  6. Human reproductive cloning and reasons for deprivation.

    Science.gov (United States)

    Jensen, D A

    2008-08-01

    Human reproductive cloning provides the possibility of genetically related children for persons for whom present technologies are ineffective. I argue that the desire for genetically related children is not, by itself, a sufficient reason to engage in human reproductive cloning. I show this by arguing that the value underlying the desire for genetically related children implies a tension between the parent and the future child. This tension stems from an instance of a deprivation and violates a general principle of reasons for deprivation. Alternative considerations, such as a right to procreative autonomy, do not appear helpful in making the case for human reproductive cloning merely on the basis of the desire for genetically related children.

  7. Quantum Nonadiabatic Cloning of Entangled Coherent States.

    Science.gov (United States)

    Izmaylov, Artur F; Joubert-Doriol, Loïc

    2017-04-20

    We propose a systematic approach to the basis set extension for nonadiabatic dynamics of entangled combination of nuclear coherent states (CSs) evolving according to the time-dependent variational principle (TDVP). The TDVP provides a rigorous framework for fully quantum nonadiabatic dynamics of closed systems; however, the quality of results strongly depends on available basis functions. Starting with a single nuclear CS replicated vertically on all electronic states, our approach clones this function when replicas of the CS on different electronic states experience increasingly different forces. Created clones move away from each other (decohere), extending the basis set. To determine a moment for cloning, we introduce generalized forces based on derivatives that maximally contribute to a variation of the total quantum action and thus account for entanglement of all basis functions.

  8. [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.

  9. Ultra-low background DNA cloning system.

    Directory of Open Access Journals (Sweden)

    Kenta Goto

    Full Text Available Yeast-based in vivo cloning is useful for cloning DNA fragments into plasmid vectors and is based on the ability of yeast to recombine the DNA fragments by homologous recombination. Although this method is efficient, it produces some by-products. We have developed an "ultra-low background DNA cloning system" on the basis of yeast-based in vivo cloning, by almost completely eliminating the generation of by-products and applying the method to commonly used Escherichia coli vectors, particularly those lacking yeast replication origins and carrying an ampicillin resistance gene (Amp(r. First, we constructed a conversion cassette containing the DNA sequences in the following order: an Amp(r 5' UTR (untranslated region and coding region, an autonomous replication sequence and a centromere sequence from yeast, a TRP1 yeast selectable marker, and an Amp(r 3' UTR. This cassette allowed conversion of the Amp(r-containing vector into the yeast/E. coli shuttle vector through use of the Amp(r sequence by homologous recombination. Furthermore, simultaneous transformation of the desired DNA fragment into yeast allowed cloning of this DNA fragment into the same vector. We rescued the plasmid vectors from all yeast transformants, and by-products containing the E. coli replication origin disappeared. Next, the rescued vectors were transformed into E. coli and the by-products containing the yeast replication origin disappeared. Thus, our method used yeast- and E. coli-specific "origins of replication" to eliminate the generation of by-products. Finally, we successfully cloned the DNA fragment into the vector with almost 100% efficiency.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. [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.

  16. Cloning arbuscule-related genes from mycorrhizas

    DEFF Research Database (Denmark)

    Burleigh, Stephen

    2000-01-01

    Until recently little was known about the identity of the genes expressed in the arbuscules of mycorrhizas, due in part to problems associated with cloning genes from the tissues of an obligate symbiont. However, the combination of advanced molecular techniques, innovative use of the materials...... available and fortuitous cloning has resulted in the recent identification of a number of arbuscule-related genes. This article provides a brief summary of the genes involved in arbuscule development, function and regulation, and the techniques used to study them. Molecular techniques include differential...

  17. Reproduction: widespread cloning in echinoderm larvae.

    Science.gov (United States)

    Eaves, Alexandra A; Palmer, A Richard

    2003-09-11

    Asexual reproduction by free-living invertebrate larvae is a rare and enigmatic phenomenon and, although it is known to occur in sea stars and brittle stars, it has not been detected in other echinoderms despite more than a century of intensive study. Here we describe spontaneous larval cloning in three species from two more echinoderm classes: a sea cucumber (Holothuroidea), a sand dollar and a sea urchin (Echinoidea). Larval cloning may therefore be an ancient ability of echinoderms and possibly of deutero-stomes - the group that includes echinoderms, acorn worms, sea squirts and vertebrates.

  18. Cloning and expression profiling of the VLDLR gene associated with egg performance in duck (Anas platyrhynchos

    Directory of Open Access Journals (Sweden)

    Peng Xiu-li

    2011-08-01

    Full Text Available Abstract Background The very low density lipoprotein receptor gene (VLDLR, a member of the low density lipoprotein receptor (LDLR gene family, plays a crucial role in the synthesis of yolk protein precursors in oviparous species. Differential splicing of this gene has been reported in human, rabbit and rat. In chicken, studies showed that the VLDLR protein on the oocyte surface mediates the uptake of yolk protein precursors into growing oocytes. However, information on the VLDLR gene in duck is still scarce. Methods Full-length duck VLDLR cDNA was obtained by comparative cloning and rapid amplification of cDNA ends (RACE. Tissue expression patterns were analysed by semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR. Association between the different genotypes and egg performance traits was investigated with the general linear model (GLM procedure of the SAS® software package. Results In duck, two VLDLR transcripts were identified, one transcript (variant-a containing an O-linked sugar domain and the other (variant-b not containing this sugar domain. These transcripts share ~70 to 90% identity with their counterparts in other species. A phylogenetic tree based on amino acid sequences showed that duck VLDLR proteins were closely related with those of chicken and zebra finch. The two duck VLDLR transcripts are differentially expressed i.e. VLDLR-a is mainly expressed in muscle tissue and VLDLR-b in reproductive organs. We have localized the duck VLDLR gene on chromosome Z. An association analysis using two completely linked SNP sites (T/C at position 2025 bp of the ORF and G/A in intron 13 and records from two generations demonstrated that the duck VLDLR gene was significantly associated with egg production (P Conclusions Duck and chicken VLDLR genes probably perform similar function in the development of growing oocytes and deposition of yolk lipoprotein. Therefore, VLDLR could be a candidate gene for duck egg performance

  19. Implications of extreme life span in clonal organisms: millenary clones in meadows of the threatened seagrass Posidonia oceanica.

    Science.gov (United States)

    Arnaud-Haond, Sophie; Duarte, Carlos M; Diaz-Almela, Elena; Marbà, Núria; Sintes, Tomas; Serrão, Ester A

    2012-01-01

    The maximum size and age that clonal organisms can reach remains poorly known, although we do know that the largest natural clones can extend over hundreds or thousands of metres and potentially live for centuries. We made a review of findings to date, which reveal that the maximum clone age and size estimates reported in the literature are typically limited by the scale of sampling, and may grossly underestimate the maximum age and size of clonal organisms. A case study presented here shows the occurrence of clones of slow-growing marine angiosperm Posidonia oceanica at spatial scales ranging from metres to hundreds of kilometres, using microsatellites on 1544 sampling units from a total of 40 locations across the Mediterranean Sea. This analysis revealed the presence, with a prevalence of 3.5 to 8.9%, of very large clones spreading over one to several (up to 15) kilometres at the different locations. Using estimates from field studies and models of the clonal growth of P. oceanica, we estimated these large clones to be hundreds to thousands of years old, suggesting the evolution of general purpose genotypes with large phenotypic plasticity in this species. These results, obtained combining genetics, demography and model-based calculations, question present knowledge and understanding of the spreading capacity and life span of plant clones. These findings call for further research on these life history traits associated with clonality, considering their possible ecological and evolutionary implications.

  20. Implications of extreme life span in clonal organisms: millenary clones in meadows of the threatened seagrass Posidonia oceanica.

    Directory of Open Access Journals (Sweden)

    Sophie Arnaud-Haond

    Full Text Available The maximum size and age that clonal organisms can reach remains poorly known, although we do know that the largest natural clones can extend over hundreds or thousands of metres and potentially live for centuries. We made a review of findings to date, which reveal that the maximum clone age and size estimates reported in the literature are typically limited by the scale of sampling, and may grossly underestimate the maximum age and size of clonal organisms. A case study presented here shows the occurrence of clones of slow-growing marine angiosperm Posidonia oceanica at spatial scales ranging from metres to hundreds of kilometres, using microsatellites on 1544 sampling units from a total of 40 locations across the Mediterranean Sea. This analysis revealed the presence, with a prevalence of 3.5 to 8.9%, of very large clones spreading over one to several (up to 15 kilometres at the different locations. Using estimates from field studies and models of the clonal growth of P. oceanica, we estimated these large clones to be hundreds to thousands of years old, suggesting the evolution of general purpose genotypes with large phenotypic plasticity in this species. These results, obtained combining genetics, demography and model-based calculations, question present knowledge and understanding of the spreading capacity and life span of plant clones. These findings call for further research on these life history traits associated with clonality, considering their possible ecological and evolutionary implications.

  1. cDNA cloning and mRNA expression of heat shock protein 70 gene ...

    African Journals Online (AJOL)

    In this study, the full-length heat shock protein 70 of Tegillarca granosa was cloned from cDNA library by rapid amplification of cDNA end (RACE). The open reading frame (ORF) of heat shock protein 70 was 1968 bp, and it encoded a protein of 655 amino acids with a predicted molecular weight of 71.48 kDa and an ...

  2. Efficient four fragment cloning for the construction of vectors for targeted gene replacement in filamentous fungi

    DEFF Research Database (Denmark)

    Frandsen, Rasmus John Normand; Andersson, Jens A.; Kristensen, Matilde Bylov

    2008-01-01

    Background: The rapid increase in whole genome fungal sequence information allows large scale functional analyses of target genes. Efficient transformation methods to obtain site-directed gene replacement, targeted over-expression by promoter replacement, in-frame epitope tagging or fusion of cod...... with an average efficiency of 84% for gene replacement and 80% for targeted overexpression. Conclusion: The new vectors designed for USER Friendly cloning provided a fast reliable method to construct vectors for targeted gene manipulations in fungi....

  3. Growth and nutrition of eucalyptus clones seedlings inoculated with mycorrhizal fungi

    OpenAIRE

    Francisco de Sousa Lima; Carla da Silva Sousa

    2014-01-01

    Eucalyptus is one of the most planted forest species, in Brazil, due to its rapid growth and high economic yield. Arbuscular mycorrhizal fungi improve the seedlings nutritional and phytosanitary status, besides increasing their resistance to biotic and abiotic stress. This study aimed to evaluate the effect of inoculation with arbuscular mycorrhizal fungi species on the growth and nutrition of different eucalyptus clones seedlings. The experiment was conducted under greenhouse conditions, in ...

  4. 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....

  5. 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

  6. Changes in the gut microbiota of cloned and non-cloned control pigs during development of obesity: gut microbiota during development of obesity in cloned pigs

    DEFF Research Database (Denmark)

    Pedersen, Rebecca; Andersen, Anders Daniel; Mølbak, Lars

    2013-01-01

    suitable for a diet-intervention study. Cloning of pigs may be an attractive way to reduce genetic influences when investigating the effect of diet and obesity on different physiological sites. The aim of this study was to assess and compare the changes in the composition of the gut microbiota of cloned vs.......0001) and in non-cloned control pigs (r=0.9, Pcomposition or variability of the fecal microbiota between the cloned pigs or between cloned and non-cloned control......Background Obesity induced by a high-caloric diet has previously been associated with changes in the gut microbiota in mice and in humans. In this study, pigs were cloned to minimize genetic and biological variation among the animals with the aim of developing a controlled metabolomic model...

  7. Cloning and characterization of functional keratinassociated protein ...

    African Journals Online (AJOL)

    Keratin-associated proteins (KAPs) 5-4 which belongs to keratin-associated protein (KRTAP) type 5 family has two major groups: high/ultrahigh cysteine (HS) and high glycine-tyrosine (HGT). Based on bioinformatic prediction, we experimentally cloned a fragment containingan open reading frame of 1849 bp from maize, ...

  8. Cloning and molecular characterization of glyceraldehyde-3 ...

    African Journals Online (AJOL)

    USER

    2010-05-31

    May 31, 2010 ... gene encoding this enzyme has been cloned and charac- terized in several filamentous fungi such as Cryphonectria parasitica (Choi and Nuss, 1990), L. edodes (Hirano et al.,. 1999), M. circinelloides (Wolff and Arnau, 2002), Phaeo- sphaeria nodorum (Ueng et al., 2003), Pleurotus sajor- caju (Jeong et al., ...

  9. Information cloning of harmonic oscillator coherent states

    Indian Academy of Sciences (India)

    Abstract. We show that in the case of unknown harmonic oscillator coherent states it is possible to achieve what we call perfect information cloning. By this we mean that it is still possible to make arbitrary number of copies of a state which has exactly the same information content as the original unknown coherent state.

  10. Cloning, expression, purification and antigenic evaluation of ...

    African Journals Online (AJOL)

    Streptococcus pyogenes produce an extracellular hyaluronidase which is associated with the spread of the organism during infection. Enzyme hyaluronidase is capable of degrading hyaluronic acid. The aim of the present study was to clone and express antigenic regions of the hylA of S.pyogenes in Escherichia coli.

  11. Cloning and characterization of functional keratin- associated ...

    African Journals Online (AJOL)

    Yomi

    2012-04-10

    Apr 10, 2012 ... Full Length Research Paper. Cloning and characterization of functional keratin- associated protein 5-4 gene in maize. Lin Yang, Feng-Ling Fu, Long-Qun Deng, Shu-Feng Zhou, Tai-Ming Yong and Wan-Chen Li*. Maize Research Institute, Sichuan Agricultural University, Chengdu, Sichuan 611130, P.R. ...

  12. No-cloning of quantum steering

    Science.gov (United States)

    Chiu, Ching-Yi; Lambert, Neill; Liao, Teh-Lu; Nori, Franco; Li, Che-Ming

    2016-06-01

    Einstein-Podolsky-Rosen (EPR) steering allows two parties to verify their entanglement, even if one party’s measurements are untrusted. This concept has not only provided new insights into the nature of non-local spatial correlations in quantum mechanics, but also serves as a resource for one-sided device-independent quantum information tasks. Here, we investigate how EPR steering behaves when one-half of a maximally entangled pair of qudits (multidimensional quantum systems) is cloned by a universal cloning machine. We find that EPR steering, as verified by a criterion based on the mutual information between qudits, can only be found in one of the copy subsystems but not both. We prove that this is also true for the single-system analogue of EPR steering. We find that this restriction, which we term ‘no-cloning of quantum steering’, elucidates the physical reason why steering can be used to secure sources and channels against cloning-based attacks when implementing quantum communication and quantum computation protocols.

  13. Information cloning of harmonic oscillator coherent states

    Indian Academy of Sciences (India)

    We show that in the case of unknown harmonic oscillator coherent statesit is possible to achieve what we call perfect information cloning. By this we mean that it is still possible to make arbitrary number of copies of a state which has exactly the same information content as the original unknown coherent state. By making use ...

  14. Cloning of partial putative gonadotropin hormone receptor ...

    Indian Academy of Sciences (India)

    Keywords. Glycoprotein hormone receptor; gonadotropin receptor; Labeo rohita; luteinizing hormone receptor; mariner transposon; PCR cloning. Abstract. A search for the presence of mariner-like elements in the Labeo rohita genome by polymerase chain reaction led to the amplification of a partial DNA sequence coding ...

  15. Cloning, expression, purification and antigenic evaluation of ...

    African Journals Online (AJOL)

    Yomi

    2012-01-31

    Jan 31, 2012 ... Key word: Hyaluronidase gene, cloning, expression of recombinant gene, antigenic region. INTRODUCTION. Group A streptococcus (Streptococcus pyogenes) is an important species of gram-positive pathogenic extra- cellular bacteria. This bacteria can produce wide range of infectious diseases like ...

  16. Cloning and expression of Phanerochaete chrysosporium ...

    African Journals Online (AJOL)

    Owner

    The Phanerochaete chrysosporium ME446 cbhI.2 cDNA was cloned and expressed in Escherichia coli ... demonstrated that two types of mRNA are produced for ... and Technology, Manchester, UK. A 2000-fold-diluted antiserum was able to detect 10 pg of the synthetic CBH peptide. cbhI.2 gene. The sequence data for P.

  17. Metabolomic phenotyping of a cloned pig model

    DEFF Research Database (Denmark)

    Clausen, Morten Rahr; Christensen, Kirstine Lykke; Hedemann, Mette Skou

    2011-01-01

    Background Pigs are widely used as models for human physiological changes in intervention studies, because of the close resemblance between human and porcine physiology and the high degree of experimental control when using an animal model. Cloned animals have, in principle, identical genotypes...

  18. Hot water and dilute acid pretreatment of high and low specific gravity Populus deltoides clones.

    Science.gov (United States)

    Martin, Elizabeth M; Bunnell, Kris A; Lau, Ching-Shuan; Pelkki, Matthew H; Patterson, David W; Clausen, Edgar C; Smith, James A; Carrier, Danielle Julie

    2011-02-01

    Populus sp. are hardwood feedstocks that grow in forest management areas that are logged for softwoods; however, they are also being considered as an energy-destined feedstock. The objective of this work was to determine the effect of xylose yield from dilute acid and hot water pretreatments performed in unstirred batch stainless steel reactors at temperatures ranging from 140 to 200°C. Populus deltoides clones S13C20 and S7C15 used in this study originated from Eastern Texas and were cultivated for 14 years in Pine Tree, AR. P. deltoides clones S13C20 and S7C15 had specific gravities of 0.48 and 0.40, respectively. Bark and wood were examined separately. As expected, hot water pretreatments, in the tested temperature range, resulted in very little direct xylose recovery. However, the 140°C dilute acid pretreatment of the lower specific gravity clone, S7C15, wood yielded the highest average xylose recovery of 56%. This condition also yielded the highest concentration of furfural, 9 mg/g sample, which can be inhibitory to the fermentation step. The highest xylose recovery from bark samples, 31%, was obtained with clone S7C15, using the 160°C dilute acid pretreatment for 60 min.

  19. An innovative strategy to clone positive modifier genes of defects caused by mtDNA mutations: MRPS18C as suppressor gene of m.3946G>A mutation in MT-ND1 gene.

    Science.gov (United States)

    Rodríguez-García, María Elena; Cotrina-Vinagre, Francisco Javier; Carnicero-Rodríguez, Patricia; Martínez-Azorín, Francisco

    2017-07-01

    We have developed a new functional complementation approach to clone modifier genes which overexpression is able to suppress the biochemical defects caused by mtDNA mutations (suppressor genes). This strategy consists in transferring human genes into respiratory chain-deficient fibroblasts, followed by a metabolic selection in a highly selective medium. We used a normalized expression cDNA library in an episomal vector (pREP4) to transfect the fibroblasts, and a medium with glutamine and devoid of any carbohydrate source to select metabolically. Growing the patient's fibroblasts in this selective medium, the deficient cells rapidly disappear unless they are rescued by the cDNA of a suppressor gene. The use of an episomal vector allows us to carry out several rounds of transfection/selection (cyclical phenotypic rescue) to enrich the rescue with true clones of suppressor genes. Using fibroblasts from a patient with epileptic encephalopathy with the m.3946G>A (p.E214K) mutation in the MT-ND1 gene, several candidate genes were identified and one of them was characterized functionally. Thus, overexpression of MRPS18C gene (that encode for bS18m protein) suppressed the molecular defects produced by this mtDNA mutation, recovering the complex I activity and reducing the ROS produced by this complex to normal levels. We suggest that modulation of bS18m expression may be an effective therapeutic strategy for the patients with this mutation.

  20. 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.

  1. A dual-site gateway cloning system for simultaneous cloning of two genes for plant transformation.

    Science.gov (United States)

    Aboulela, Mostafa; Tanaka, Yuji; Nishimura, Kohji; Mano, Shoji; Kimura, Tetsuya; Nakagawa, Tsuyoshi

    2017-07-01

    Analyses of the subcellular localization of proteins and protein-protein interaction networks are essential to uncover the molecular basis of diverse biological processes in plants. To this end, we have created a Gateway cloning-compatible vector system, named dual-site (DS) Gateway cloning system to allow simple cloning of two expression cassettes in a binary vector and to express them simultaneously in plant cells. In the DS Gateway cloning system, (i) a moderate constitutive nopaline synthase promoter (Pnos), which is much suitable for localization analysis, is used to guide each expression cassette, (ii) four series of vectors with different plant resistance markers are established, (iii) N-terminal fusion with 6 fluorescent proteins and 7 epitope tags is available, (iv) both N- and C-terminal fusions with split enhanced yellow fluorescent protein (EYFP) are possible for efficient detection of protein-protein interactions using a bimolecular fluorescence complementation (BiFC) assay. The usefulness of the DS Gateway cloning system has been demonstrated by the analysis of the expression and the subcellular localization patterns of two Golgi proteins in stable expression system using A. thaliana, and by the analyses of interactions between subunits of coat protein complex II (COPII) both in transient and stable expression systems using Japanese leek and A. thaliana, respectively. The DS Gateway cloning system provides a multipurpose, efficient expression tool in gene function analyses and especially suitable for investigating interactions and subcellular localization of two proteins in living plant cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system.

    Science.gov (United States)

    Sasaki, Yukari; Sone, Takefumi; Yoshida, Shouhei; Yahata, Kazuhide; Hotta, Junko; Chesnut, Jonathan D; Honda, Takeshi; Imamoto, Fumio

    2004-02-05

    Six types of recombination signal DNA sequences of the Multisite Gateway cloning system were investigated as to their specificity and efficiency in the LR and BP recombination reactions. In the LR reaction to generate an Expression clone by recombination between attL and attR signals which are contained in the Entry clone and the Destination vector, respectively, the cross-reactivity of various attL and attR pairs on six types of respective signal sequences was examined. In the BP reaction to create an Entry clone by transferring the target DNA segment in the Expression clone or the attB-flanked PCR product into a Donor vector, various combinations of attB and attP pairs were tested for their reactivities in recombination. The results obtained indicate a markedly higher specificity and efficiency of cross-reactivity with only the matched att signal pairs, such as attL3-attR3, attB5-attP5, and so on, compared to unmatched signal pairs, such as attL3-attR5, attB5-attP3, and so on, thus verifying a high-throughput production of the positive clones in the Gateway system in which multiple recombination signals exist together in one reaction system. Examples of rapid construction of a three or four DNA-fusion structure in the plasmid are shown.

  3. Assessment of genetic diversity in different clones of Dalbergia ...

    African Journals Online (AJOL)

    Jane

    2011-07-13

    cluster II consisted maximum of twelve clones followed by cluster I (11 clones) and cluster VI (7 clones) whereas clusters. IV, VII and VIII consisted .... Bhatinda Branch Canal 70-71 L/S, Malerkotla, Sangrur, Punjab. 30° 12' N.

  4. Technological Literacy and Human Cloning. Resources in Technology.

    Science.gov (United States)

    Baird, Steven L.

    2002-01-01

    Discusses how technology educators can deal with advances in human genetics, specifically, cloning. Includes a definition and history of cloning, discusses its benefits, and looks at social concerns and arguments for and against human cloning. Includes classroom activities and websites. (Contains 10 references.) (JOW)

  5. Distribution of quantum Fisher information in asymmetric cloning machines.

    Science.gov (United States)

    Xiao, Xing; Yao, Yao; Zhou, Lei-Ming; Wang, Xiaoguang

    2014-12-08

    An unknown quantum state cannot be copied and broadcast freely due to the no-cloning theorem. Approximate cloning schemes have been proposed to achieve the optimal cloning characterized by the maximal fidelity between the original and its copies. Here, from the perspective of quantum Fisher information (QFI), we investigate the distribution of QFI in asymmetric cloning machines which produce two nonidentical copies. As one might expect, improving the QFI of one copy results in decreasing the QFI of the other copy. It is perhaps also unsurprising that asymmetric phase-covariant cloning outperforms universal cloning in distributing QFI since a priori information of the input state has been utilized. However, interesting results appear when we compare the distributabilities of fidelity (which quantifies the full information of quantum states), and QFI (which only captures the information of relevant parameters) in asymmetric cloning machines. Unlike the results of fidelity, where the distributability of symmetric cloning is always optimal for any d-dimensional cloning, we find that any asymmetric cloning outperforms symmetric cloning on the distribution of QFI for d ≤ 18, whereas some but not all asymmetric cloning strategies could be worse than symmetric ones when d > 18.

  6. Molecular cloning, characterization and expression of WAG-2 ...

    Indian Academy of Sciences (India)

    Gel Extraction kit (Qiagen, Shanghai, China). Purified PCR products were cloned in a pMD20T vector following the man- ufacturer's instructions (Takara, Dalian, China). Transfor- mants were plated on LB agar containing ampicillin. Clones. Table 1. Primers and probes for WAG-2 cloning and expression from Ae. tauschii.

  7. Molecular cloning of endochitinase 33 (ECH33) gene from ...

    African Journals Online (AJOL)

    Jane

    2011-09-28

    Sep 28, 2011 ... This study was conducted to screen for the presence of ech33 gene in 80 isolates of Trichoderma. Furthermore, using gene specific primers, ech33 gene were cloned into pTZ57R/T from T. harzanium. IABT1068. The clone was confirmed through PCR amplification and restriction analysis. The clones.

  8. A Gateway MultiSite recombination cloning toolkit.

    Directory of Open Access Journals (Sweden)

    Lena K Petersen

    Full Text Available The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org.

  9. Early selection of Eucalyptus clones in retrospective nursery test ...

    African Journals Online (AJOL)

    Within the framework of the eucalyptus breeding programme in the Congo, two retrospective tests were conducted using mature clones in the field and young cuttings under nursery conditions with two hybrids: 13 clones of Eucalyptus tereticornis* Eucalyptus grandis for the test TC 82-1B and 17 clones of Eucalyptus ...

  10. A Gateway MultiSite recombination cloning toolkit.

    Science.gov (United States)

    Petersen, Lena K; Stowers, R Steven

    2011-01-01

    The generation of DNA constructs is often a rate-limiting step in conducting biological experiments. Recombination cloning of single DNA fragments using the Gateway system provided an advance over traditional restriction enzyme cloning due to increases in efficiency and reliability. Here we introduce a series of entry clones and a destination vector for use in two, three, and four fragment Gateway MultiSite recombination cloning whose advantages include increased flexibility and versatility. In contrast to Gateway single-fragment cloning approaches where variations are typically incorporated into model system-specific destination vectors, our Gateway MultiSite cloning strategy incorporates variations in easily generated entry clones that are model system-independent. In particular, we present entry clones containing insertions of GAL4, QF, UAS, QUAS, eGFP, and mCherry, among others, and demonstrate their in vivo functionality in Drosophila by using them to generate expression clones including GAL4 and QF drivers for various trp ion channel family members, UAS and QUAS excitatory and inhibitory light-gated ion channels, and QUAS red and green fluorescent synaptic vesicle markers. We thus establish a starter toolkit of modular Gateway MultiSite entry clones potentially adaptable to any model system. An inventory of entry clones and destination vectors for Gateway MultiSite cloning has also been established (www.gatewaymultisite.org).

  11. Market share for semen and cloned embryos in dairy herds.

    NARCIS (Netherlands)

    Boer, de I.J.M.; Arendonk, van J.A.M.

    1994-01-01

    Use of cloned embryos from desirable genotypes (commercial clone lines) enables faster dissemination of superior genetics to dauy producers. Under optimal purchasing strategies of milk producers, the annual proportion of replacement cows from commercial clone lines indicates the market share of

  12. 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.

  13. Population genetics of cancer cell clones: possible implications of cancer stem cells

    Directory of Open Access Journals (Sweden)

    Naugler Christopher T

    2010-11-01

    Full Text Available Abstract Background The population dynamics of the various clones of cancer cells existing within a tumour is complex and still poorly understood. Cancer cell clones can be conceptualized as sympatric asexual species, and as such, the application of theoretical population genetics as it pertains to asexual species may provide additional insights. Results The number of generations of tumour cells within a cancer has been estimated at a minimum of 40, but high cancer cell mortality rates suggest that the number of cell generations may actually be in the hundreds. Such a large number of generations would easily allow natural selection to drive clonal evolution assuming that selective advantages of individual clones are within the range reported for free-living animal species. Tumour cell clonal evolution could also be driven by variation in the intrinsic rates of increase of different clones or by genetic drift. In every scenario examined, the presence of cancer stem cells would require lower selection pressure or less variation in intrinsic rates of increase. Conclusions The presence of cancer stem cells may result in more rapid clonal evolution. Specific predictions from theoretical population genetics may lead to a greater understanding of this process.

  14. Productivitatea clonelor de plop hibrid instalate în culturi intensive în nord-estul României [ Poplar clones productivity managed for biomass production in North-Eastern Romania

    Directory of Open Access Journals (Sweden)

    Dănilă I.C.

    2016-08-01

    Full Text Available Poplar (Populus spp. is a fast-growing species in temperate conditions, with potential to substitute fossil fuels by obtaining energy from biomass. The aim of this work was to study the productivity of 6 hybrid poplar clone from a hilly region of NE Romania, after a growing season of 4 and, respectively, 5 years. Rods were used as planting material, planted at a density of 2667 trees per ha, with between-row spacing of 3 m and interior-row distances of 1.25 m. Generally, significant differences appear between clones, for all analysed biometric characteristics (p≤0.05. Results show that, in the fifth growing season, diameter increases on average with 15.1%, reaching 11.89 cm, height increases in average with 13.9%, reaching 11.89 m, and volume increases with 33.3%, reaching 0.172 m3. The total biomass after 4 years vegetation varies from 32.8 t/ha to 39.4 t/ha, and after 5 years from 47.7 t/ha to 60.2 t/ha, having an average increase in the latest growing season of 35% and an average yield of 11.3 t/ha/year. The most productive clones in the given growing conditions and crop characteristics are the clone Pannonia after 4 growing seasons and the clone AF6 after 5 years.

  15. Recent progress and problems in animal cloning.

    Science.gov (United States)

    Tsunoda, Y; Kato, Y

    2002-01-01

    It is remarkable that mammalian somatic cell nuclei can form whole individuals if they are transferred to enucleated oocytes. Advancements in nuclear transfer technology can now be applied for genetic improvement and increase of farm animals, rescue of endangered species, and assisted reproduction and tissue engineering in humans. Since July 1998, more than 200 calves have been produced by nuclear transfer of somatic cell nuclei in Japan, but half of them were stillborn or died within several months of parturition. Morphologic abnormalities have also been observed in cloned calves and embryonic stem cell-derived mice. In this review, we discuss the present situation and problems with animal cloning and the possibility for its application to human medicine.

  16. Micropropagation of an elite Darjeeling tea clone.

    Science.gov (United States)

    Jha, T; Sen, S K

    1992-03-01

    Shoot cultures of Camellia sinensis (L.) O. Kuntz var. T-78, an elite Darjeeling tea clone, were established from cotyledonary nodes and shoot tips of germinated seedlings as well as from nodal explants of field grown plants. Shoot multiplication rate ranged from 4x in nodal explants to 35x in cotyledonary nodes after 18 weeks of culture. Rooting was achieved in 80-90% micro-shoots by either placing them on an inductive medium for 10 d and then transferring shoots to hormone-free medium, or by treating micro-shoots with a chronic dose of IBA (500 mg/l) for 30-40 min. Rooted plants were established in soil under glasshouse condition at 60% frequency after hardening phase of 4-6 weeks. The regenerated plants show a constant chromosome number of 2n=30 and are morphologically true to type. This procedure can be applied for conservation and utilisation of an elite clone of Darjeeling tea.

  17. Gamer's Facial Cloning for Online Interactive Games

    Directory of Open Access Journals (Sweden)

    Abdul Sattar

    2009-01-01

    Full Text Available Virtual illustration of a human face is essential to enhance the mutual interaction in a cyber community. In this paper we propose a solution to solve two bottlenecks in facial analysis and synthesis for an interactive system of human face cloning for non-expert users of computer games. Tactical maneuvers of the gamer make single camera acquisition system unsuitable to analyze and track the face due to its large lateral movements. For an improved facial analysis system, we propose to acquire the facial images from multiple cameras and analyze them by multiobjective 2.5D Active Appearance Model (MOAAM. Facial morphological dissimilarities between a human face and an avatar make the facial synthesis quite complex. To successfully clone or retarget the gamer facial expressions and gestures on to an avatar, we introduce a simple mathematical link between their appearances. Results obtained validate the efficiency, accuracy and robustness achieved.

  18. Generating in vivo cloning vectors for parallel cloning of large gene clusters by homologous recombination.

    Directory of Open Access Journals (Sweden)

    Jeongmin Lee

    Full Text Available A robust method for the in vivo cloning of large gene clusters was developed based on homologous recombination (HR, requiring only the transformation of PCR products into Escherichia coli cells harboring a receiver plasmid. Positive clones were selected by an acquired antibiotic resistance, which was activated by the recruitment of a short ribosome-binding site plus start codon sequence from the PCR products to the upstream position of a silent antibiotic resistance gene in receiver plasmids. This selection was highly stringent and thus the cloning efficiency of the GFPuv gene (size: 0.7 kb was comparable to that of the conventional restriction-ligation method, reaching up to 4.3 × 10(4 positive clones per μg of DNA. When we attempted parallel cloning of GFPuv fusion genes (size: 2.0 kb and carotenoid biosynthesis pathway clusters (sizes: 4 kb, 6 kb, and 10 kb, the cloning efficiency was similarly high regardless of the DNA size, demonstrating that this would be useful for the cloning of large DNA sequences carrying multiple open reading frames. However, restriction analyses of the obtained plasmids showed that the selected cells may contain significant amounts of receiver plasmids without the inserts. To minimize the amount of empty plasmid in the positive selections, the sacB gene encoding a levansucrase was introduced as a counter selection marker in receiver plasmid as it converts sucrose to a toxic levan in the E. coli cells. Consequently, this method yielded completely homogeneous plasmids containing the inserts via the direct transformation of PCR products into E. coli cells.

  19. The potential for modification in cloning and vitrification technology to enhance genetic progress in beef cattle in Northern Australia.

    Science.gov (United States)

    Taylor-Robinson, Andrew W; Walton, Simon; Swain, David L; Walsh, Kerry B; Vajta, Gábor

    2014-08-01

    Recent advances in embryology and related research offer considerable possibilities to accelerate genetic improvement in cattle breeding. Such progress includes optimization and standardization of laboratory embryo production (in vitro fertilization - IVF), introduction of a highly efficient method for cryopreservation (vitrification), and dramatic improvement in the efficiency of somatic cell nuclear transfer (cloning) in terms of required effort, cost, and overall outcome. Handmade cloning (HMC), a simplified version of somatic cell nuclear transfer, offers the potential for relatively easy and low-cost production of clones. A potentially modified method of vitrification used at a centrally located laboratory facility could result in cloned offspring that are economically competitive with elite animals produced by more traditional means. Apart from routine legal and intellectual property issues, the main obstacle that hampers rapid uptake of these technologies by the beef cattle industry is a lack of confidence from scientific and commercial sources. Once stakeholder support is increased, the combined application of these methods makes a rapid advance toward desirable traits (rapid growth, high-quality beef, optimized reproductive performance) a realistic goal. The potential impact of these technologies on genetic advancement in beef cattle herds in which improvement of stock is sought, such as in northern Australia, is hard to overestimate. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Keeping up with the cloneses--issues in human cloning.

    Science.gov (United States)

    Rollin, B E

    1999-01-01

    The advent of cloning animals has created a maelstrom of social concern about the "ethical issues" associated with the possibility of cloning humans. When the "ethical concerns" are clearly examined, however, many of them turn out to be less matters of rational ethics than knee-jerk emotion, religious bias, or fear of that which is not understood. Three categories of real and spurious ethical concerns are presented and discussed: 1) that cloning is intrinsically wrong, 2) that cloning must lead to bad consequences, and 3) that cloning harms the organism generated. The need for a rational ethical framework for discussing biotechnological advances is presented and defended.

  1. Cloning humans? Biological, ethical, and social considerations.

    Science.gov (United States)

    Ayala, Francisco J

    2015-07-21

    There are, in mankind, two kinds of heredity: biological and cultural. Cultural inheritance makes possible for humans what no other organism can accomplish: the cumulative transmission of experience from generation to generation. In turn, cultural inheritance leads to cultural evolution, the prevailing mode of human adaptation. For the last few millennia, humans have been adapting the environments to their genes more often than their genes to the environments. Nevertheless, natural selection persists in modern humans, both as differential mortality and as differential fertility, although its intensity may decrease in the future. More than 2,000 human diseases and abnormalities have a genetic causation. Health care and the increasing feasibility of genetic therapy will, although slowly, augment the future incidence of hereditary ailments. Germ-line gene therapy could halt this increase, but at present, it is not technically feasible. The proposal to enhance the human genetic endowment by genetic cloning of eminent individuals is not warranted. Genomes can be cloned; individuals cannot. In the future, therapeutic cloning will bring enhanced possibilities for organ transplantation, nerve cells and tissue healing, and other health benefits.

  2. Pathogenicity of the highly leukotoxic JP2 clone of Aggregatibacter actinomycetemcomitans and its geographic dissemination and role in aggressive periodontitis

    Directory of Open Access Journals (Sweden)

    Dorte Haubek

    2014-08-01

    Full Text Available For decades, Aggregatibacter actinomycetemcomitans has been associated with aggressive forms of periodontitis in adolescents. In the middle of the 1990s, a specific JP2 clone of A. actinomycetemcomitans, belonging to the cluster of serotype b strains of A. actinomycetemcomitans and having a number of other characteristics, was found to be strongly associated with aggressive forms of periodontitis, particularly in North Africa. Although several longitudinal studies still point to the bacterial species, A. actinomycetemcomitans as a risk factor of aggressive periodontitis, it is now also widely accepted that the highly leukotoxic JP2 clone of A. actinomycetemcomitans is implicated in rapidly progressing forms of aggressive periodontitis. The JP2 clone strains are highly prevalent in human populations living in Northern and Western parts of Africa. These strains are also prevalent in geographically widespread populations that have originated from the Northwest Africa. Only sporadic signs of a dissemination of the JP2 clone strains to non-African populations have been found despite Africans living geographically widespread for hundreds of years. It remains an unanswered question if a particular host tropism exists as a possible explanation for the frequent colonization of the Northwest African population with the JP2 clone. Two exotoxins of A. actinomycetemcomitans are known, leukotoxin (LtxA and cytolethal distending toxin (Cdt. LtxA is able to kill human immune cells, and Cdt can block cell cycle progression in eukaryotic cells and thus induce cell cycle arrest. Whereas the leukotoxin production is enhanced in JP2 clone strains thus increasing the virulence potential of A. actinomycetemcomitans, it has not been possible so far to demonstrate such a role for Cdt. Lines of evidence have led to the understanding of the highly leukotoxic JP2 clone of A. actinomycetemcomitans as an aetiological factor of aggressive periodontitis. Patients, who are

  3. Pathogenicity of the highly leukotoxic JP2 clone of Aggregatibacter actinomycetemcomitans and its geographic dissemination and role in aggressive periodontitis

    Science.gov (United States)

    Haubek, Dorte; Johansson, Anders

    2014-01-01

    For decades, Aggregatibacter actinomycetemcomitans has been associated with aggressive forms of periodontitis in adolescents. In the middle of the 1990s, a specific JP2 clone of A. actinomycetemcomitans, belonging to the cluster of serotype b strains of A. actinomycetemcomitans and having a number of other characteristics, was found to be strongly associated with aggressive forms of periodontitis, particularly in North Africa. Although several longitudinal studies still point to the bacterial species, A. actinomycetemcomitans as a risk factor of aggressive periodontitis, it is now also widely accepted that the highly leukotoxic JP2 clone of A. actinomycetemcomitans is implicated in rapidly progressing forms of aggressive periodontitis. The JP2 clone strains are highly prevalent in human populations living in Northern and Western parts of Africa. These strains are also prevalent in geographically widespread populations that have originated from the Northwest Africa. Only sporadic signs of a dissemination of the JP2 clone strains to non-African populations have been found despite Africans living geographically widespread for hundreds of years. It remains an unanswered question if a particular host tropism exists as a possible explanation for the frequent colonization of the Northwest African population with the JP2 clone. Two exotoxins of A. actinomycetemcomitans are known, leukotoxin (LtxA) and cytolethal distending toxin (Cdt). LtxA is able to kill human immune cells, and Cdt can block cell cycle progression in eukaryotic cells and thus induce cell cycle arrest. Whereas the leukotoxin production is enhanced in JP2 clone strains thus increasing the virulence potential of A. actinomycetemcomitans, it has not been possible so far to demonstrate such a role for Cdt. Lines of evidence have led to the understanding of the highly leukotoxic JP2 clone of A. actinomycetemcomitans as an aetiological factor of aggressive periodontitis. Patients, who are colonized with the JP2

  4. Ethical issues regarding human cloning: a nursing perspective.

    Science.gov (United States)

    Dinç, Leyla

    2003-05-01

    Advances in cloning technology and successful cloning experiments in animals raised concerns about the possibility of human cloning in recent years. Despite many objections, this is not only a possibility but also a reality. Human cloning is a scientific revolution. However, it also introduces the potential for physical and psychosocial harm to human beings. From this point of view, it raises profound ethical, social and health related concerns. Human cloning would have an impact on the practice of nursing because it could result in the creation of new physiological and psychosocial conditions that would require nursing care. The nursing profession must therefore evaluate the ethics of human cloning, in particular the potential role of nurses. This article reviews the ethical considerations of reproductive human cloning, discusses the main reasons for concern, and reflects a nursing perspective regarding this issue.

  5. U.S. consumers attitudes toward farm animal cloning.

    Science.gov (United States)

    Brooks, Kathleen R; Lusk, Jayson L

    2011-10-01

    In January 2008, the United States Food and Drug Administration concluded "meat and milk from cattle, swine, and goat clones or their offspring are as safe to eat as food we eat from those species now" (U.S. FDA, 2010). However, cloning remains a very controversial topic. A web-based survey administered by Knowledge Networks was used to determine U.S. consumers' awareness of and attitudes toward meat and milk from cloned cattle. Findings reveal consumers do not differentiate much between products from cloned animals and products from non-cloned animals. Overall consumers are concerned that animal cloning is an unnatural process and that it will lead to human cloning. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. [DNA methylation and development abnormalities in cloned animals].

    Science.gov (United States)

    Yang, Rong-Rong; Li, Xiang-Yun

    2007-09-01

    Most cloned animals by nuclear transfer were dead before their births, and only a few can develop to their late gestation or adulthood. Although some cloned offsprings can survive, they often have some development disfigurements and abnormal phenotypes in various degrees. DNA methylation is an important modifiable manner of epigenetic dominating the correct expression of gene. It is a main instrument of regulating genome function and plays a prominent part in the embryonic normal development. Through researching the pattern of DNA methylation, we found that there were many abnormal DNA methylation patterns in cloned animals, which might be the primary reasons for inducing premature death of cloned embryos and development abnormalities of cloned animals. This article discusses the function of DNA methylation, the aberrant DNA methylation patterns in cloned animals, and the reasons of inducing abnormal DNA methylation in cloned animals.

  7. The effect of water availability on stand-level productivity, transpiration, water use efficiency and radiation use efficiency of field-grown willow clones

    DEFF Research Database (Denmark)

    Linderson, Maj-Lena; Iritz, Z.; Lindroth, A.

    2007-01-01

    The effect of water availability on stand-level productivity, transpiration, water use efficiency (WUE) and radiation use efficiency (RUE) is evaluated for different willow clones at stand level. The measurements were made during the growing season 2000 in a 3-year-old plantation in Scania...... low compared to other results. Generally, all clones, except for Jorunn, seem to be better off concerning biomass production, WUE and RUE than the reference clone. Jorr, Jorunn and Loden also seem to be able to cope with the reduced water availability with increase in the water use efficiency. Tora...... performs significantly better than the other clones concerning both growth and efficiency in light and water use, but the effect of the dry treatment on stem growth shows sensitivity to water availability. The reduced stem growth could be due to a change in allocation patterns. (C) 2007 Elsevier Ltd. All...

  8. Long-adapter single-strand oligonucleotide probes for the massively multiplexed cloning of kilobase genome regions.

    Science.gov (United States)

    Tosi, Lorenzo; Sridhara, Viswanadham; Yang, Yunlong; Guan, Dongli; Shpilker, Polina; Segata, Nicola; Larman, H Benjamin; Parekkadan, Biju

    2017-01-01

    As the catalogue of sequenced genomes and metagenomes continues to grow, massively parallel approaches for the comprehensive and functional analysis of gene products and regulatory elements are becoming increasingly valuable. Current strategies to synthesize or clone complex libraries of DNA sequences are limited by the length of the DNA targets, throughput and cost. Here, we show that long-adapter single-strand oligonucleotide (LASSO) probes can capture and clone thousands of kilobase DNA fragments in a single reaction. As a proof-of-principle, we simultaneously cloned >3,000 bacterial open reading frames (ORFs) from E. coli genomic DNA (spanning 400-5,000 bp targets). Targets were enriched up to a median of ~60-fold compared to non-targeted genomic regions. At a cutoff of 3 times the median non-target reads per kilobase of genetic element per million reads, ~75% of the targeted ORFs were successfully captured. We also show that LASSO probes can clone human ORFs from complementary DNA, and an ORF library from a human-microbiome sample. LASSO probes could be used for the preparation of long-read sequencing libraries and for massively multiplexed cloning.

  9. 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.

  10. (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, ...

  11. [Genetic polymorphism of clones and their seed progeny in the scotch pine clone plantation].

    Science.gov (United States)

    Korshikov, I I; Demkovich, A E

    2010-01-01

    Genetic variation at 12 allozyme loci (10 of them being polymorphic ones) has been studied in the archive-clone plantation of 23 Pinus sylvestris plus-trees and their seed progeny in the south-east of Ukraine. More than a half of clones had 4-8 heterozygous loci, whereas their seed progeny was marked by a lower variation than maternal trees. Seed progeny was obtained at a high outcrossing rate (t(m) = 95%). The clone progeny was characterized by a high percentage of abnormal allele segregation in megagametophytes. There was also a high frequency of significant deviation in distribution of seed embryo genotypes from the theoretically expected one according to the Hardy-Weinberg law.

  12. Boron mobility in eucalyptus clones Mobilidade de Boro em clones de eucalipto

    Directory of Open Access Journals (Sweden)

    Jackson Freitas Brilhante de São José

    2009-12-01

    Full Text Available Understanding the magnitude of B mobility in eucalyptus may help to select clones that are more efficient for B use and to design new practices of B fertilization. This study consisted of five experiments with three eucalyptus clones (129, 57 and 58 where the response to and mobility of B were evaluated. Results indicated that clone 129 was less sensitive to B deficiency than clones 68 and 57, apparently due to its ability to translocate B previously absorbed via root systems to younger tissues when B in solution became limiting. Translocation also occurred when B was applied as boric acid only once to a single mature leaf, resulting in higher B concentration in roots, stems and younger leaves. The growth of B-deficient plants was also recovere by a single foliar application of B to a mature leaf. This mobility was greater, when foliar-applied B was supplied in complexed (boric acid + manitol than in non-complexed form (boric acid alone. When the root system of clone 129 was split in two solution compartments, B supplied to one root compartment was translocated to the shoot and back to the roots in the other compartment, improving the B status and growth. Thus, it appears that B is relatively mobile in eucalyptus, especially in clone 129, and its higher mobility could be due to the presence of an organic compound such as manitol, able to complex B.O entendimento da magnitude da mobilidade de B no eucalipto é importante, devido a sua utilidade para a seleção de clones mais eficiente para esse micronutriente, além da projeção de novas práticas de fertilização de B. Este estudo envolveu cinco experimentos com três clones de eucalipto (129, 57 e 68, em que foram avaliadas a resposta e a mobilidade de B. Os resultados indicaram que o clone 129 foi menos sensível à deficiência de B do que os clones 68 e 57, aparentemente, em razão da sua maior habilidade de translocar B, previamente absorvido via sistema radicular, para tecidos jovens

  13. 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.

  14. Transgenic and cloned animals in the food chain--are we prepared to tackle it?

    Science.gov (United States)

    Jagadeesan, Premanandh; Bin Salem, Samara

    2015-11-01

    Transgenic and cloned animal production for various purposes has been increasing rapidly in recent times. While the actual impact of these animals in the food chain is unknown, the significance of tracking and monitoring measures to curb accidental and or deliberate release has been discussed. Religious perspectives from different faiths and traditions have been presented. Although the concept of substantial equivalence satisfies the technical and nutritional requirements of these products when assessed against comparators, public opinion and religious concerns should also be considered by the regulators while developing policy regulations. In conclusion, measures to prevent real or perceived risks of transgenic and cloned animals in food production require global coordinated action. It is worthwhile to consider establishing effective tracking systems and analytical procedures as this will be a valuable tool if a global consensus is not reached on policy regulation. © 2015 Society of Chemical Industry.

  15. Apoptosis Gene Hunting Using Retroviral Expression Cloning: Identification of Vacuolar ATPase Subunit E

    Directory of Open Access Journals (Sweden)

    Claire L. Anderson

    2003-01-01

    Full Text Available Over the past 10-15 years there has been an explosion of interest in apoptosis. The delayed realisation that cell death is an essential part of life for any multicellular organism has meant that, despite the recent and rapid developments of the last decade, the precise biochemical pathways involved in apoptosis remain incomplete and potentially novel genes may, as yet, remain undiscovered. The hunt is therefore on to bridge the remaining gaps in our knowledge. Our contribution to this research effort utilises a functional cloning approach to isolate important regulatory genes involved in apoptosis. This mini-review focuses on the use and advantages of a retroviral expression cloning strategy and describes the isolation and identification of one such potential apoptosis regulatory gene, namely that encoding vacuolar ATPase subunit E.

  16. Phylogeography: English elm is a 2,000-year-old Roman clone.

    Science.gov (United States)

    Gil, Luis; Fuentes-Utrilla, Pablo; Soto, Alvaro; Cervera, M Teresa; Collada, Carmen

    2004-10-28

    The outbreak of Dutch elm disease in the 1970s ravaged European elm populations, killing more than 25 million trees in Britain alone; the greatest impact was on Ulmus procera, otherwise known as the English elm. Here we use molecular and historical information to show that this elm derives from a single clone that the Romans transported from Italy to the Iberian peninsula, and from there to Britain, for the purpose of supporting and training vines. Its highly efficient vegetative reproduction and its inability to set seeds have preserved this clone unaltered for 2,000 years as the core of the English elm population--and the preponderance of this susceptible variety may have favoured a rapid spread of the disease.

  17. Should we clone human beings? Cloning as a source of tissue for transplantation.

    Science.gov (United States)

    Savulescu, J

    1999-01-01

    The most publicly justifiable application of human cloning, if there is one at all, is to provide self-compatible cells or tissues for medical use, especially transplantation. Some have argued that this raises no new ethical issues above those raised by any form of embryo experimentation. I argue that this research is less morally problematic than other embryo research. Indeed, it is not merely morally permissible but morally required that we employ cloning to produce embryos or fetuses for the sake of providing cells, tissues or even organs for therapy, followed by abortion of the embryo or fetus. PMID:10226910

  18. Human cloning: category, dignity, and the role of bioethics.

    Science.gov (United States)

    Shuster, Evelyne

    2003-10-01

    Human cloning has been simultaneously a running joke for massive worldwide publicity of fringe groups like the Raelians, and the core issue of an international movement at the United Nations in support of a treaty to ban the use of cloning techniques to produce a child (so called reproductive cloning). Yet, even though debates on human cloning have greatly increased since the birth of Dolly, the clone sheep, in 1997, we continue to wonder whether cloning is after all any different from other methods of medically assisted reproduction, and what exactly makes cloning an 'affront to the dignity of humans.' Categories we adopt matter mightily as they inform but can also misinform and lead to mistaken and unproductive decisions. And thus bioethicists have a responsibility to ensure that the proper categories are used in the cloning debates and denounce those who try to win the ethical debate through well-crafted labels rather than well-reasoned argumentations. But it is as important for bioethicists to take a position on broad issues such as human cloning and species altering interventions. One 'natural question' would be, for example, should there be an international treaty to ban human reproductive cloning?

  19. Procreative liberty, enhancement and commodification in the human cloning debate.

    Science.gov (United States)

    Shapshay, Sandra

    2012-12-01

    The aim of this paper is to scrutinize a contemporary standoff in the American debate over the moral permissibility of human reproductive cloning in its prospective use as a eugenic enhancement technology. I shall argue that there is some significant and under-appreciated common ground between the defenders and opponents of human cloning. Champions of the moral and legal permissibility of cloning support the technology based on the right to procreative liberty provided it were to become as safe as in vitro fertilization and that it be used only by adults who seek to rear their clone children. However, even champions of procreative liberty oppose the commodification of cloned embryos, and, by extension, the resulting commodification of the cloned children who would be produced via such embryos. I suggest that a Kantian moral argument against the use of cloning as an enhancement technology can be shown to be already implicitly accepted to some extent by champions of procreative liberty on the matter of commodification of cloned embryos. It is in this argument against commodification that the most vocal critics of cloning such as Leon Kass and defenders of cloning such as John Robertson can find greater common ground. Thus, I endeavor to advance the debate by revealing a greater degree of moral agreement on some fundamental premises than hitherto recognized.

  20. High-dimensional quantum cloning and applications to quantum hacking.

    Science.gov (United States)

    Bouchard, Frédéric; Fickler, Robert; Boyd, Robert W; Karimi, Ebrahim

    2017-02-01

    Attempts at cloning a quantum system result in the introduction of imperfections in the state of the copies. This is a consequence of the no-cloning theorem, which is a fundamental law of quantum physics and the backbone of security for quantum communications. Although perfect copies are prohibited, a quantum state may be copied with maximal accuracy via various optimal cloning schemes. Optimal quantum cloning, which lies at the border of the physical limit imposed by the no-signaling theorem and the Heisenberg uncertainty principle, has been experimentally realized for low-dimensional photonic states. However, an increase in the dimensionality of quantum systems is greatly beneficial to quantum computation and communication protocols. Nonetheless, no experimental demonstration of optimal cloning machines has hitherto been shown for high-dimensional quantum systems. We perform optimal cloning of high-dimensional photonic states by means of the symmetrization method. We show the universality of our technique by conducting cloning of numerous arbitrary input states and fully characterize our cloning machine by performing quantum state tomography on cloned photons. In addition, a cloning attack on a Bennett and Brassard (BB84) quantum key distribution protocol is experimentally demonstrated to reveal the robustness of high-dimensional states in quantum cryptography.

  1. Human somatic cell nuclear transfer and cloning.

    Science.gov (United States)

    2012-10-01

    This document presents arguments that conclude that it is unethical to use somatic cell nuclear transfer (SCNT) for infertility treatment due to concerns about safety; the unknown impact of SCNT on children, families, and society; and the availability of other ethically acceptable means of assisted reproduction. This document replaces the ASRM Ethics Committee report titled, "Human somatic cell nuclear transfer (cloning)," last published in Fertil Steril 2000;74:873-6. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  2. Insights on bovine genetic engineering and cloning

    Directory of Open Access Journals (Sweden)

    Fabiana F. Bressan

    2013-12-01

    Full Text Available Transgenic technology has become an essential tool for the development of animal biotechnologies, and animal cloning through somatic cell nuclear transfer (SCNT enabled the generation of genetically modified animals utilizing previously modified and selected cell lineages as nuclei donors, assuring therefore the generation of homogeneous herds expressing the desired modification. The present study aimed to discuss the use of SCNT as an important methodology for the production of transgenic herds, and also some recent insights on genetic modification of nuclei donors and possible effects of gene induction of pluripotency on SCNT.

  3. Quantum correlations support probabilistic pure state cloning

    Energy Technology Data Exchange (ETDEWEB)

    Roa, Luis, E-mail: lroa@udec.cl [Departamento de Física, Universidad de Concepción, Casilla 160-C, Concepción (Chile); Alid-Vaccarezza, M.; Jara-Figueroa, C. [Departamento de Física, Universidad de Concepción, Casilla 160-C, Concepción (Chile); Klimov, A.B. [Departamento de Física, Universidad de Guadalajara, Avenida Revolución 1500, 44420 Guadalajara, Jalisco (Mexico)

    2014-02-01

    The probabilistic scheme for making two copies of two nonorthogonal pure states requires two auxiliary systems, one for copying and one for attempting to project onto the suitable subspace. The process is performed by means of a unitary-reduction scheme which allows having a success probability of cloning different from zero. The scheme becomes optimal when the probability of success is maximized. In this case, a bipartite state remains as a free degree which does not affect the probability. We find bipartite states for which the unitarity does not introduce entanglement, but does introduce quantum discord between some involved subsystems.

  4. Effects of donor fibroblast cell type and transferred cloned embryo number on the efficiency of pig cloning.

    Science.gov (United States)

    Li, Zicong; Shi, Junsong; Liu, Dewu; Zhou, Rong; Zeng, Haiyu; Zhou, Xiu; Mai, Ranbiao; Zeng, Shaofen; Luo, Lvhua; Yu, Wanxian; Zhang, Shouquan; Wu, Zhenfang

    2013-02-01

    Currently, cloning efficiency in pigs is very low. Donor cell type and number of cloned embryos transferred to an individual surrogate are two major factors that affect the successful rate of somatic cell nuclear transfer (SCNT) in pigs. This study aimed to compare the influence of different donor fibroblast cell types and different transferred embryo numbers on recipients' pregnancy rate and delivery rate, the average number of total clones born, clones born alive and clones born healthy per litter, and the birth rate of healthy clones (=total number of healthy cloned piglets born /total number of transferred cloned embryos). Three types of donor fibroblasts were tested in large-scale production of cloned pigs, including fetal fibroblasts (FFBs) from four genetically similar Western swine breeds of Pietrain (P), Duroc (D), Landrace (L), and Yorkshire (Y), which are referred to as P,D,LY-FFBs, adult fibroblasts (AFBs) from the same four breeds, which are designated P,D,L,Y-AFBs, and AFBs from a Chinese pig breed of Laiwu (LW), which is referred to as LW-AFBs. Within each donor fibroblast cell type group, five transferred cloned embryo number groups were tested. In each embryo number group, 150-199, 200-249, 250-299, 300-349, or 350-450 cloned embryos were transferred to each individual recipient sow. For the entire experiment, 92,005 cloned embryos were generated from nearly 115,000 matured oocytes and transferred to 328 recipients; in total, 488 cloned piglets were produced. The results showed that the mean clones born healthy per litter resulted from transfer of embryos cloned from LW-AFBs (2.53 ± 0.34) was similar with that associated with P,D,L,Y-FFBs (2.72 ± 0.29), but was significantly higher than that resulted from P,D,L,Y-AFBs (1.47 ± 0.18). Use of LW-AFBs as donor cells for SCNT resulted in a significantly higher pregnancy rate (72.00% vs. 59.30% and 48.11%) and delivery rate (60.00% vs. 45.93% and 35.85%) for cloned embryo recipients, and a

  5. Protective properties of radio-chemoresistant glioblastoma stem cell clones are associated with metabolic adaptation to reduced glucose dependence.

    Directory of Open Access Journals (Sweden)

    Fei Ye

    Full Text Available Glioblastoma stem cells (GSC are a significant cell model for explaining brain tumor recurrence. However, mechanisms underlying their radiochemoresistance remain obscure. Here we show that most clonogenic cells in GSC cultures are sensitive to radiation treatment (RT with or without temozolomide (TMZ. Only a few single cells survive treatment and regain their self-repopulating capacity. Cells re-populated from treatment-resistant GSC clones contain more clonogenic cells compared to those grown from treatment-sensitive GSC clones, and repeated treatment cycles rapidly enriched clonogenic survival. When compared to sensitive clones, resistant clones exhibited slower tumor development in animals. Upregulated genes identified in resistant clones via comparative expression microarray analysis characterized cells under metabolic stress, including blocked glucose uptake, impaired insulin/Akt signaling, enhanced lipid catabolism and oxidative stress, and suppressed growth and inflammation. Moreover, many upregulated genes highlighted maintenance and repair activities, including detoxifying lipid peroxidation products, activating lysosomal autophagy/ubiquitin-proteasome pathways, and enhancing telomere maintenance and DNA repair, closely resembling the anti-aging effects of caloric/glucose restriction (CR/GR, a nutritional intervention that is known to increase lifespan and stress resistance in model organisms. Although treatment-introduced genetic mutations were detected in resistant clones, all resistant and sensitive clones were subclassified to either proneural (PN or mesenchymal (MES glioblastoma subtype based on their expression profiles. Functional assays demonstrated the association of treatment resistance with energy stress, including reduced glucose uptake, fatty acid oxidation (FAO-dependent ATP maintenance, elevated reactive oxygen species (ROS production and autophagic activity, and increased AMPK activity and NAD(+ levels accompanied by

  6. Consumers' attitudes toward consumption of cloned beef. The impact of exposure to technological information about animal cloning.

    Science.gov (United States)

    Aizaki, Hideo; Sawada, Manabu; Sato, Kazuo

    2011-10-01

    Novel food technologies, such as cloning, have been introduced into the meat production sector; however, their use is not widely supported by many consumers. This study was designed to assess whether Japanese consumers' attitudes toward consumption of cloned beef (specifically, beef derived from bovine embryo and somatic cell-cloned cattle) would change after they were provided with technological information on animal cloning through a web-based survey. The results revealed that most respondents did not discriminate between their attitudes toward the consumption of the two types of cloned beef, and that most respondents did not change their attitudes toward cloned beef after receiving the technological information. The respondents' individual characteristics, including their knowledge about the food safety of cloned beef and their basic knowledge about animal cloning, influenced the likelihood of a change in their attitudes after they received the information. In conclusion, some consumers might become less uncomfortable about the consumption of cloned beef by the straightforward provision of technological information about animal cloning; however, most consumers are likely to maintain their attitudes. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy

    Directory of Open Access Journals (Sweden)

    Bi Yanzhen

    2012-07-01

    Full Text Available Abstract Background Widely used restriction-dependent cloning methods are labour-intensive and time-consuming, while several types of ligase-independent cloning approaches have inherent limitations. A rapid and reliable method of cloning native DNA sequences into desired plasmids are highly desired. Results This paper introduces ABI-REC, a novel strategy combining asymmetric bridge PCR with intramolecular homologous recombination in bacteria for native DNA cloning. ABI-REC was developed to precisely clone inserts into defined location in a directional manner within recipient plasmids. It featured an asymmetric 3-primer PCR performed in a single tube that could robustly amplify a chimeric insert-plasmid DNA sequence with homologous arms at both ends. Intramolecular homologous recombination occurred to the chimera when it was transformed into E.coli and produced the desired recombinant plasmids with high efficiency and fidelity. It is rapid, and does not involve any operational nucleotides. We proved the reliability of ABI-REC using a double-resistance reporter assay, and investigated the effects of homology and insert length upon its efficiency. We found that 15 bp homology was sufficient to initiate recombination, while 25 bp homology had the highest cloning efficiency. Inserts up to 4 kb in size could be cloned by this method. The utility and advantages of ABI-REC were demonstrated through a series of pig myostatin (MSTN promoter and terminator reporter plasmids, whose transcriptional activity was assessed in mammalian cells. We finally used ABI-REC to construct a pig MSTN promoter-terminator cassette reporter and showed that it could work coordinately to express EGFP. Conclusions ABI-REC has the following advantages: (i rapid and highly efficient; (ii native DNA cloning without introduction of extra bases; (iii restriction-free; (iv easy positioning of directional and site-specific recombination owing to formulated primer design. ABI

  8. An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy.

    Science.gov (United States)

    Bi, Yanzhen; Qiao, Xianfeng; Hua, Zaidong; Zhang, Liping; Liu, Ximei; Li, Li; Hua, Wenjun; Xiao, Hongwei; Zhou, Jingrong; Wei, Qingxin; Zheng, Xinmin

    2012-07-06

    Widely used restriction-dependent cloning methods are labour-intensive and time-consuming, while several types of ligase-independent cloning approaches have inherent limitations. A rapid and reliable method of cloning native DNA sequences into desired plasmids are highly desired. This paper introduces ABI-REC, a novel strategy combining asymmetric bridge PCR with intramolecular homologous recombination in bacteria for native DNA cloning. ABI-REC was developed to precisely clone inserts into defined location in a directional manner within recipient plasmids. It featured an asymmetric 3-primer PCR performed in a single tube that could robustly amplify a chimeric insert-plasmid DNA sequence with homologous arms at both ends. Intramolecular homologous recombination occurred to the chimera when it was transformed into E.coli and produced the desired recombinant plasmids with high efficiency and fidelity. It is rapid, and does not involve any operational nucleotides. We proved the reliability of ABI-REC using a double-resistance reporter assay, and investigated the effects of homology and insert length upon its efficiency. We found that 15 bp homology was sufficient to initiate recombination, while 25 bp homology had the highest cloning efficiency. Inserts up to 4 kb in size could be cloned by this method. The utility and advantages of ABI-REC were demonstrated through a series of pig myostatin (MSTN) promoter and terminator reporter plasmids, whose transcriptional activity was assessed in mammalian cells. We finally used ABI-REC to construct a pig MSTN promoter-terminator cassette reporter and showed that it could work coordinately to express EGFP. ABI-REC has the following advantages: (i) rapid and highly efficient; (ii) native DNA cloning without introduction of extra bases; (iii) restriction-free; (iv) easy positioning of directional and site-specific recombination owing to formulated primer design. ABI-REC is a novel approach to DNA engineering and gene functional

  9. Preservation and Reproduction of Microminipigs by Cloning Technology.

    Science.gov (United States)

    Enya, Satoko; Kawarasaki, Tatsuo; Otake, Masayoshi; Kangawa, Akihisa; Uenishi, Hirohide; Mikawa, Satoshi; Nishimura, Takashi; Kuwahawa, Yasushi; Shibata, Masatoshi

    Microminipigs have been maintained in small populations of closed colonies, involving risks of inbreeding depression and genetic drift. In order to avoid these risks, we assessed the applicability of cloning technology. Male and female clones were produced from a stock of cryopreserved somatic cells, obtaining offspring by means of natural mating. Phenotypic and genotypic characteristics of original microminipigs, clones and their offspring were analyzed and recorded. Clones presented characteristics similar to those of the cell-stock data. Although the body weight of clones tended to be heavier than that of the cell-stock data, body weights of their offspring were similar to those of previous reports. Thus, cloned microminipigs have the potential to be a valuable genetic resource for reproduction and breeding. Our proposed methodology might be useful to provide a large number of animals with adequate quality from a limited population with sufficient genetic diversity. Copyright © 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  10. Cloning animals by somatic cell nuclear transfer – biological factors

    Science.gov (United States)

    Tian, X Cindy; Kubota, Chikara; Enright, Brian; Yang, Xiangzhong

    2003-01-01

    Cloning by nuclear transfer using mammalian somatic cells has enormous potential application. However, somatic cloning has been inefficient in all species in which live clones have been produced. High abortion and fetal mortality rates are commonly observed. These developmental defects have been attributed to incomplete reprogramming of the somatic nuclei by the cloning process. Various strategies have been used to improve the efficiency of nuclear transfer, however, significant breakthroughs are yet to happen. In this review we will discuss studies conducted, in our laboratories and those of others, to gain a better understanding of nuclear reprogramming. Because cattle are a species widely used for nuclear transfer studies, and more laboratories have succeeded in cloning cattle than any other specie, this review will be focused on somatic cell cloning of cattle. PMID:14614770

  11. Willow yield is highly dependent on clone and site

    DEFF Research Database (Denmark)

    Ugilt Larsen, Søren; Jørgensen, Uffe; Lærke, Poul Erik

    2014-01-01

    , differing considerably in soil type, climatic conditions and management. Compared to the best clone, the yield was up to 36 % lower for other clones across sites and up to 51 % lower within sites. Tordis was superior to other clones with dry matter yields between 5.2 and 10.2 Mg ha−1 year−1 during the first...... to the best site, yield level was up to 51 % lower on other sites across all clones, probably due to combined effects of differences in soil type, climate and management. Thus, willow yield depends both on the use of high-yielding clones and on the combined site effects of soil, climate, and management.......Use of high-yielding genotypes is one of the means to achieve high yield and profitability in willow (Salix spp.) short rotation coppice. This study investigated the performance of eight willow clones (Inger, Klara, Linnea, Resolution, Stina, Terra Nova, Tora, Tordis) on five Danish sites...

  12. Early Evaluation of Promising White Poplar (Populus alba L. Clones in Hungary

    Directory of Open Access Journals (Sweden)

    RÉDEI, Károly

    2010-01-01

    Full Text Available White poplar (Populus alba L. and its natural hybrid, grey poplar (Populus x canescens are native stand-forming tree species in Hungary, covering 3.4 per cent of the forested area (64 000 ha. More than 70 per cent of their stands and plantations can be found on calcareous sandy sites in the Danube–Tisza region, so they play a significant role in the poplar growing of this part of the country. One of the most important tasks ahead of Hungarian poplar growers is to improve the quality of poplar stands and plantations for wood production based on selecting relatively drought-tolerant clones and cultivars. In the paper the juvenile growth and the morphological characteristics of four micropropagated white poplar clones have been evaluated on a marginal site in central Hungary. The clones ‘H-337’ (P. alba x P. grandidentata and ‘H-384‘ (P. alba x P. grandidentata appear to be especially promising for quality wood production under arid hydrological conditions.

  13. Ecological significance of assimilate distribution in Agropyron repens clones under influence of the copper smelter Legnica

    Directory of Open Access Journals (Sweden)

    Teresa Brej

    2014-01-01

    Full Text Available The studies on couch grass (Agropyron repens (L. P. Beauv. populations growing in stress conditions in close vicinity of a copper smelter concern the integrity of clonal structure. The connections of tillers within a clone and the interclonal integrity was investigated by means of assimilate translocation, using 14C. It was found that heavy metal stress affects the phenotypic plasticity of couch grass in regard to clonal growth of the phalanx type. The phalanx type growth is supported by a considerable integration, which allows the redistribution of resources, through internal routes, from places rich in resources to such clone parts, which cover a surface poor in nutritive compounds. In an unpolluted (control couch grass population representing the guerilla type of growth, the translocation of assimilates concerns only the closest (sister ramets. The phalanx type of growth in couch grass subjected to contamination favours also the accumulation of organic matter and macronutrients (N, Ca in zones of occurrence of Agropyron repens clusters, which are in deficit in areas close to the smelter. The irregular accumulation of heavy metals and the deficit of macronutrients in soil, form near the smelter a patchy environment. In this patchy environment couch grass, as one of few plants, finds appropriate conditions for foraging. The whole of factors in the studied polluted area creates a unique dynamic system between couch grass clones and the local ecological conditions.

  14. Productivity, leaf traits and carbon isotope discrimination in 29 Populus deltoides x P. nigra clones.

    Science.gov (United States)

    Monclus, Romain; Dreyer, Erwin; Delmotte, Francis M; Villar, Marc; Delay, Didier; Boudouresque, Eric; Petit, Jean-Michel; Marron, Nicolas; Bréchet, Claude; Brignolas, Franck

    2005-07-01

    Here we tested whether some leaf traits could be used as predictors for productivity in a range of Populus deltoides x P. nigra clones. These traits were assessed in 3-yr-old rooted cuttings from 29 clones growing in an open field trial, in a five randomized complete block design, under optimal irrigation. Variables were assigned to four groups describing productivity (above-ground biomass, total leaf area), leaf growth (total number of leaves increment rate), leaf structure (area of the largest leaf, specific leaf area, carbon and nitrogen contents), and carbon isotope discrimination in the leaves (Delta). High-yielding clones displayed larger total leaf area and individual leaf area, while no correlation could be detected between productivity and either leaf structure or Delta. By contrast, Delta was negatively correlated with number of leaves increment rate and leaf N content. Our study shows that there is a potential to improve water-use efficiency in poplar without necessarily reducing the overall productivity.

  15. Growing America's Energy Future

    Energy Technology Data Exchange (ETDEWEB)

    None

    2016-06-01

    The emerging U.S. bioenergy industry provides a secure and growing supply of transportation fuels, biopower, and bioproducts produced from a range of abundant, renewable biomass resources. Bioenergy can help ensure a secure, sustainable, and economically sound future by reducing U.S. dependence on foreign oil, developing domestic clean energy sources, and generating domestic green jobs. Bioenergy can also help address growing concerns about climate change by reducing greenhouse gas emissions to create a healthier environment for current and future generations.

  16. Rapid cloning of genes in hexaploid wheat using cultivar-specific long-range chromosome assembly

    Czech Academy of Sciences Publication Activity Database

    Thind, A. K.; Wicker, T.; Šimková, Hana; Fossati, D.; Moullet, O.; Brabant, C.; Vrána, Jan; Doležel, Jaroslav; Krattinger, S.G.

    2017-01-01

    Roč. 35, č. 8 (2017), s. 793-796 ISSN 1087-0156 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : adult-plant resistance * leaf rust * sequence capture * genome * arabidopsis * virulence * barley * canada * locus * lr22a Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 41.667, year: 2016

  17. Rapid cloning of disease-resistance genes in plants using mutagenesis and sequence capture

    Science.gov (United States)

    Genetic solutions to protect crops against pests and pathogens are preferable to agrichemicals 1. Wild crop relatives carry immense diversity of disease resistance (R) genes that could enable more sustainable disease control. However, recruiting R genes for crop improvement typically involves long b...

  18. Determining Complementary Properties with Quantum Clones

    Science.gov (United States)

    Thekkadath, G. S.; Saaltink, R. Y.; Giner, L.; Lundeen, J. S.

    2017-08-01

    In a classical world, simultaneous measurements of complementary properties (e.g., position and momentum) give a system's state. In quantum mechanics, measurement-induced disturbance is largest for complementary properties and, hence, limits the precision with which such properties can be determined simultaneously. It is tempting to try to sidestep this disturbance by copying the system and measuring each complementary property on a separate copy. However, perfect copying is physically impossible in quantum mechanics. Here, we investigate using the closest quantum analog to this copying strategy, optimal cloning. The coherent portion of the generated clones' state corresponds to "twins" of the input system. Like perfect copies, both twins faithfully reproduce the properties of the input system. Unlike perfect copies, the twins are entangled. As such, a measurement on both twins is equivalent to a simultaneous measurement on the input system. For complementary observables, this joint measurement gives the system's state, just as in the classical case. We demonstrate this experimentally using polarized single photons.

  19. [Cloning: necessary reflections on the imaginary].

    Science.gov (United States)

    Minahim, María Auxiliadora

    2009-01-01

    The article covers the innumerable reasons given for using cloning for therapeutic and reproductive purposes. The most commonly used argument in favour of the procedure has been that of preserving human dignity, which would include the wide exercising of personal autonomy without restrictions of an ethical nature. This view is countered by questions relating to the use of the technique, namely self-determination and the loss of the integrity of the species, which would include the transformation of a generation through the production of human beings and tissues. It must also be made clear that therapeutic cloning (which is carried out through the use of stem cells) is not yet a reality in the scientific world, with the result that the procedure that is supposedly necessary, which argues in favour of the destruction of the young embryo is misleading, as are also certain discourses used to refer to the theme and the science. Criminal law, on prohibiting this practice is anticipating it becoming a reality, protecting legal rights that affect supra-individual interests, such as the destruction of the young embryo, one of the issues of concern to ADIN (Acción Directa de Inconstitucionalidad en Brasil - Direct Action on Unconstitutionality in Brazil) 3510-0.

  20. Towards an understanding of British public attitudes concerning human cloning.

    Science.gov (United States)

    Shepherd, Richard; Barnett, Julie; Cooper, Helen; Coyle, Adrian; Moran-Ellis, Jo; Senior, Victoria; Walton, Chris

    2007-07-01

    The ability of scientists to apply cloning technology to humans has provoked public discussion and media coverage. The present paper reports on a series of studies examining public attitudes to human cloning in the UK, bringing together a range of quantitative and qualitative methods to address this question. These included a nationally representative survey, an experimental vignette study, focus groups and analyses of media coverage. Overall the research presents a complex picture of attitude to and constructions of human cloning. In all of the analyses, therapeutic cloning was viewed more favourably than reproductive cloning. However, while participants in the focus groups were generally negative about both forms of cloning, and this was also reflected in the media analyses, quantitative results showed more positive responses. In the quantitative research, therapeutic cloning was generally accepted when the benefits of such procedures were clear, and although reproductive cloning was less accepted there was still substantial support. Participants in the focus groups only differentiated between therapeutic and reproductive cloning after the issue of therapeutic cloning was explicitly raised; initially they saw cloning as being reproductive cloning and saw no real benefits. Attitudes were shown to be associated with underlying values associated with scientific progress rather than with age, gender or education, and although there were a few differences in the quantitative data based on religious affiliation, these tended to be small effects. Likewise in the focus groups there was little direct appeal to religion, but the main themes were 'interfering with nature' and the 'status of the embryo', with the latter being used more effectively to try to close down further discussion. In general there was a close correspondence between the media analysis and focus group responses, possibly demonstrating the importance of media as a resource, or that the media reflect

  1. [Human cloning and the protection of women's interests].

    Science.gov (United States)

    Canabes, Marcela Ahumada

    2008-01-01

    The Human Cloning, both therapeutic and full birth cloning, involves and affects women in a special way. The United Nation's Declaration on the Cloning of Human Beings includes a special clause referred to them. Also the Spanish law does it. This works pretend to analyse the meaning of the inclusion of women's interests in this document. At the same time, I will consider the foundations and the importance of the reference to the women.

  2. Growth performance and variability of Dalbergia sissoo Roxb. clones

    OpenAIRE

    K.R. Dhixya Deve* and K.T. Parthiban

    2014-01-01

    The study was undertaken at Forest College and Research Institute, Mettupalayam to ascertain the growth performance and variability of Dalbergia sissoo clones. Twenty clones collected from plus trees in different parts of India were assessed for three characters namely collar diameter, height and volume index. Based on the growth performance at six months after planting, five clones namely FCRIDSC 3, FCRIDSC 4, FCRIDSC 7, FCRIDSC 8 and FCRIDSC 16 were found be superior. GCV, heritability and ...

  3. Grizzly bear corticosteroid binding globulin: Cloning and serum protein expression.

    Science.gov (United States)

    Chow, Brian A; Hamilton, Jason; Alsop, Derek; Cattet, Marc R L; Stenhouse, Gordon; Vijayan, Mathilakath M

    2010-06-01

    Serum corticosteroid levels are routinely measured as markers of stress in wild animals. However, corticosteroid levels rise rapidly in response to the acute stress of capture and restraint for sampling, limiting its use as an indicator of chronic stress. We hypothesized that serum corticosteroid binding globulin (CBG), the primary transport protein for corticosteroids in circulation, may be a better marker of the stress status prior to capture in grizzly bears (Ursus arctos). To test this, a full-length CBG cDNA was cloned and sequenced from grizzly bear testis and polyclonal antibodies were generated for detection of this protein in bear sera. The deduced nucleotide and protein sequences were 1218 bp and 405 amino acids, respectively. Multiple sequence alignments showed that grizzly bear CBG (gbCBG) was 90% and 83% identical to the dog CBG nucleotide and amino acid sequences, respectively. The affinity purified rabbit gbCBG antiserum detected grizzly bear but not human CBG. There were no sex differences in serum total cortisol concentration, while CBG expression was significantly higher in adult females compared to males. Serum cortisol levels were significantly higher in bears captured by leg-hold snare compared to those captured by remote drug delivery from helicopter. However, serum CBG expression between these two groups did not differ significantly. Overall, serum CBG levels may be a better marker of chronic stress, especially because this protein is not modulated by the stress of capture and restraint in grizzly bears. Copyright 2010 Elsevier Inc. All rights reserved.

  4. Fundamental resource-allocating model in colleges and universities based on Immune Clone Algorithms

    Science.gov (United States)

    Ye, Mengdie

    2017-05-01

    In this thesis we will seek the combination of antibodies and antigens converted from the optimal course arrangement and make an analogy with Immune Clone Algorithms. According to the character of the Algorithms, we apply clone, clone gene and clone selection to arrange courses. Clone operator can combine evolutionary search and random search, global search and local search. By cloning and clone mutating candidate solutions, we can find the global optimal solution quickly.

  5. Breeding of fast growing forest tree species for biomass production in Greece

    Energy Technology Data Exchange (ETDEWEB)

    Aravanopoulos, F.A. [Laboratory of Forest Genetics and Tree Breeding, Faculty of Forestry and Natural Environment, PO Box 238, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece)

    2010-11-15

    The purpose of this review is to provide a critical analysis of the literature regarding the breeding of fast growing forest tree species dedicated to biomass production in Greece and to identify future goals. The main genera employed in pertinent research which show the most promising results for operational plantations belong to Populus spp. and Platanus spp. The best poplar clone (P. x euramericana I-455) produced 16.54 t ha{sup -1} y{sup -1} dry weight averaged over different experimental plantations and rotation periods, while the best clone derived from the Greek breeding program (Populus deltoides var. missouriensis x Populus nigra var. pubescens He-X/3) produced 14.23 t ha{sup -1} y{sup -1} respectively. The best results of Platanus species and hybrids were at the vicinity of 10 t ha{sup -1} y{sup -1}. Clones selected under a combination of higher temperature and drier condition regimes reflecting the Mediterranean field conditions will be valuable in future breeding under a changing climate and increased energy needs. The high genetic diversity of the virtually unattached local genetic reserve can provide even from the initial breeding cycles great opportunities for genetic improvement and significant genetic gains. (author)

  6. Knowledge and attitudes toward human cloning in Israel.

    Science.gov (United States)

    Barnoy, Sivia; Ehrenfeld, Malka; Sharon, Rina; Tabak, Nili

    2006-04-01

    The success of mammal cloning in 1997 has brought the issue of human cloning into public discussion. Human cloning has several aspects and potential applications for use in both reproductive and non-reproductive matters. The aim of this study was to evaluate the knowledge and attitudes toward human cloning in Israel. Data from 120 respondents (68 health professionals and 52 non-health professionals), all Jewish, Hebrew speaking with at least 15 years of education each, were collected using two questionnaires that dealt with knowledge and attitudes toward human cloning. Results showed that although health professionals had significantly more knowledge that non-health professionals, all respondents had poor knowledge about cloning. No difference in attitudes was found between the groups. Most respondents opposed human cloning, but more positive attitudes toward non-reproductive cloning were found. The results are discussed in the context of the deficit model. The findings indicate a need to provide information about human cloning to allow people to form their attitudes based on factual knowledge.

  7. The inadequacies of absolute prohibition of reproductive cloning.

    Science.gov (United States)

    Lee, Martin Lishexian

    2004-02-01

    This study reviews debates on human cloning and its benefits, considers international and domestic laws, and argues that the choice of reproductive means is a human right. In exercise of this right, a balanced approach should be adopted, in order to benefit human society while protecting human dignity adequately. The immaturity of cloning techniques indicates that at the present time human reproductive cloning is too risky. Thus a temporary ban on such cloning is appropriate, but the ban on relevant scientific research and animal experimentation is inappropriate as it denies the spirit of freedom of scientific inquiry, and hinders making the benefits of scientific advancement available to human society as a whole.

  8. [Temporal layers of the clone. Remarks on a conceptual history].

    Science.gov (United States)

    Brandt, Christina

    2010-06-01

    This paper aims at a history of the clone concept in 20th-century life science and culture. The first part of the paper is concerned with conceptual history approaches. Here, the idea of 'Zeitschichten' by Reinhart Koselleck is discussed and its implications for the history of science are explored. In the following parts of the paper, I trace the historical dynamics of the clone concept in various fields of 20th-century life sciences. I argue that the clone concept, which originated in plant breeding around 1900, soon developed into a technical tool in a variety of research areas. With this, specific meanings became attached: the idea of standardization, genetic identity, and mass reproduction. A further connotation of the clone was the idea of stagnancy with respect to processes in time: The clone was seen as something that was exempt from evolutionary changes. In the last section of the paper, I trace the shifting meanings of the clone concept in the 1960s and 1970s, when the clone became a widespread metaphor that pointed to future biotechnologically driven possibilities to reshape the nature of human beings. In this regard, the debates of the 1970s are analyzed as a turning point: Whereas utopian and eugenic visions predominated the debates in the 1960s (when the human clone was seen as something which will occur in a distant future), the 1970s discussion focused on the advent of a biotechnological era and the human clone had became a reality.

  9. Counterfactual quantum cloning without transmitting any physical particles

    Science.gov (United States)

    Guo, Qi; Zhai, Shuqin; Cheng, Liu-Yong; Wang, Hong-Fu; Zhang, Shou

    2017-11-01

    We propose a counterfactual 1 →2 economical phase-covariant cloning scheme. Compared with the existing protocols using flying qubits, the main difference of the presented scheme is that the cloning can be achieved without transmitting the photon between the two parties. In addition, this counterfactual scheme does not need to construct controlled quantum gates to perform joint logical operations between the cloned qubit and the blank copy. We also numerically evaluate the performance of the present scheme in the practical experiment, which shows this cloning scheme can be implemented with a high success of probability and the fidelity is close to the optimal value in the ideal asymptotic limit.

  10. Isolation, cloning and molecular characterization of a thermotolerant ...

    African Journals Online (AJOL)

    Isolation, cloning and molecular characterization of a thermotolerant xylanase from Streptomyces sp. THW31. Thayat Sriyapai, Peechapack Somyoonsap, Supatra Areekit, Paisarn Khawsak, Arda Pakpitcharoen, Kosum Chansiri ...

  11. Clone Detection for Graph-Based Model Transformation Languages

    DEFF Research Database (Denmark)

    Strüber, Daniel; Plöger, Jennifer; Acretoaie, Vlad

    2016-01-01

    Cloning is a convenient mechanism to enable reuse across and within software artifacts. On the downside, it is also a practice related to significant long-term maintainability impediments, thus generating a need to identify clones in affected artifacts. A large variety of clone detection techniques...... has been proposed for programming and modeling languages; yet no specific ones have emerged for model transformation languages. In this paper, we explore clone detection for graph-based model transformation languages. We introduce potential use cases for such techniques in the context of constructive...

  12. Human reproductive cloning and biotechnology: Rational, ethical and public concerns

    Directory of Open Access Journals (Sweden)

    M.E.S. (Elbie van den Berg

    2012-11-01

    Full Text Available Previous research indicates a lack of clear international guidelines on the permissibility of embryonic stem cell research and human reproductive cloning. These studies suggest that this is the result of severe criticism from uninformed publics, whose arguments are based on misconceptions influenced by popular literature and science fiction films. However, the current research argues that public cloning attitudes that are based on real social and ethical concerns should be deployed to direct social and legal policy-making on human reproductive cloning. Addressing public concerns about human reproductive cloning is essential in exploring sound avenues for sensible biotechnology and policy-making. The research, on which this article reported, intended to give a critical evaluation of some major arguments for and against human reproductive cloning in order to establish whether or not these arguments hold up well under rational interrogation. Notwithstanding the author’s critical attitude to uninformed opinions, false assumptions and unsound conclusions about the complex issue of human reproductive cloning, the author argued from the perspective that every life phenomenon is inextricably intertwined with everything else, and part of larger complex webs of interactions. Such a perspective recognised that the well-being of other human beings, including future human clones, is not only an existential, social and moral imperative but also epistemological. Against the backdrop of this perspective, critical questions arose that justified the creation of human clones in the face of possible defects and abnormalities in cloned children, as well as the possible harm to societies.

  13. Rapidly- growing firms and their main characteristics: a longitudinal study from United States

    DEFF Research Database (Denmark)

    Keen, Christian; Etemad, Hamid

    2011-01-01

    This paper explores the topic of high-growth phenomenon and its agents. High-growth enterprises are primarily small and medium sized firms that attain very high-growth rates for at least five years. The paper presents a review of the pertinent literature to guide its formulation of hypotheses...... are relatively smaller enterprises and their high growth rates are not restricted to a particular location, industrial region, size or time period. The findings of this analysis point to a population of high-growth enterprises with diverse locations, sizes and times with important implications for scholarly...

  14. THE IMPORTANCE OF LEGAL ORGANISATION OF RAPIDLY GROWING COMPANIES FOR INTERNATIONAL BUSINESS COMPETITIVENESS

    Directory of Open Access Journals (Sweden)

    Riko Novak

    2014-09-01

    Full Text Available This article discusses the importance of a company’s legal form for the process of internationalisation using a sample of 1577 Slovenian companies. We refer to previous studies and on the basis of additional statistical data evaluate whether the choice of corporate legal structure influences a company’s ability to compete internationally. In the domestic market, most companies operate as limited liability companies; this is also the most frequent legal form in which companies enter foreign markets. We conclude that the form by itself does not influence the decision to go international.

  15. [Case report: Rapidly growing abdominal wall giant desmoid tumour during pregnancy].

    Science.gov (United States)

    Palacios-Zertuche, Jorge Tadeo; Cardona-Huerta, Servando; Juárez-García, María Luisa; Valdés-Flores, Everardo; Muñoz-Maldonado, Gerardo Enrique

    Desmoid tumours are one of the rarest tumours worldwide, with an estimated yearly incidence of 2-4 new cases per million people. They are soft tissue monoclonal neoplasms that originate from mesenchymal stem cells. It seems that the hormonal and immunological changes occurring during pregnancy may play a role in the severity and course of the disease. The case is presented on 28-year-old female in her fifth week of gestation, in whom an abdominal wall tumour was found attached to left adnexa and uterus while performing a prenatal ultrasound. The patient was followed up under clinical and ultrasonographic surveillance. When she presented with abnormal uterine activity at 38.2 weeks of gestation, she was admitted and obstetrics decided to perform a caesarean section. Tumour biopsy was taken during the procedure. Histopathology reported a desmoid fibromatosis. A contrast enhanced abdominal computed tomography scan was performed, showing a tumour of 26×20.5×18cm, with well-defined borders in contact with the uterus, left adnexa, bladder and abdominal wall, with no evidence of infiltration to adjacent structures. A laparotomy, with tumour resection, hysterectomy and left salpingo-oophorectomy, components separation techniques, polypropylene mesh insertion, and drainage was performed. The final histopathology report was desmoid fibromatosis. There is no evidence of recurrence after 6 months follow-up. Desmoid tumours are locally aggressive and surgical resection with clear margins is the basis for the treatment of this disease, using radiotherapy, chemotherapy and hormone therapy as an adjunct in the treatment. Copyright © 2016 Academia Mexicana de Cirugía A.C. Publicado por Masson Doyma México S.A. All rights reserved.

  16. 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.

  17. Isolation of rapid growing mycobacteria from soil and water in Iran

    African Journals Online (AJOL)

    USER

    2010-06-14

    Jun 14, 2010 ... However, laboratory protocols that are commonly used to investigate the presence of mycobacteria in clinical specimens are insufficient for isolation of these organisms form soil and natural water samples (Chilima et al., 2006). In spite of large –scale BCG trial conducted in our country during past decades, ...

  18. Case series: Rapidly growing squamous cell carcinoma after cutaneous surgical intervention

    Directory of Open Access Journals (Sweden)

    Abdullah Ibrahim

    2017-12-01

    This case series explores the various mechanisms of de-novo squamous cell carcinoma development in areas of cutaneous surgical intervention, including graft harvest. It also provides recommendations regarding the necessary precautions to avoid implantation of squamous cell carcinoma into distant sites. Lastly it highlights the importance of surveillance for any suspicious lesions arising from areas of previous cutaneous surgical intervention.

  19. Emerging Development Pathways of Urban Livestock Production in Rapidly Growing West Africa Cities

    Directory of Open Access Journals (Sweden)

    Regina Roessler

    2016-11-01

    Full Text Available In this study, we try to capture the degree of specialization or integration, and of intensification or extensification, of (peri- urban livestock production, along with the factors that influence such decisions and their impact on natural resource uses. A total of 181 and 187 structured questionnaires were completed in livestock-keeping households in Ouagadougou (Burkina Faso and Tamale (Ghana. Categorical principal component and two-step cluster analysis were used to identify homogenous groups of livestock-keeping households. Cross tabulation and logistic regression analysis revealed factors that influence livestock husbandry, showing their impacts on resource use by livestock keepers in the two cities. A diversity of livestock species was kept, mostly integrated with crop farming. Yet, some households specialized in either sheep, pig or commercial milk production, and partly intensified their production. The decision to specialize and/or intensify livestock production is site-specific and influenced by the education level of the household head and security of land ownership. Higher inputs in livestock systems do not necessarily lead to higher outputs, and specialization inevitably leads to higher manure wastages. Therefore, links of livestock producers to crop farmers and markets for livestock manure must be strengthened to enable recycling of resources and limit negative externalities of specialized livestock production. Strategies need to be identified to improve livestock productivity by enhancing outputs as input use increases.

  20. Scar sarcoidosis on a finger mimicking a rapidly growing soft tissue tumour: a case report

    Directory of Open Access Journals (Sweden)

    Henrichs Marcel-Philipp

    2012-10-01

    Full Text Available Abstract Background Scar sarcoidosis is a rare and uncommon but specific cutaneous manifestation of sarcoidosis. In general it arises in pre-existing scars deriving from mechanical traumas. As most surgeons dealing with scars might not be aware of cutaneous sarcoidosis and its different types of appearance the appropriate staging and treatment might be missed or at least delayed. To our knowledge this is the first case in literature of scar sarcoidosis on a finger. Case presentation We present a case of a 33-year-old carpenter who developed scar sarcoidosis on his right index finger 4 years after the tendon of the long digital flexor got accidentally cut by an angle grinder. He was referred due to a swelling of the finger suspected to be a malignant soft tissue tumour. The circumference of the affected finger had almost doubled, adding up to 94 mm. Incision biopsy revealed typical noncaseating granulomas. Further investigation showed a systemic extent of the disease with involvement of the lung. A systemic treatment with oral steroids led to an almost full regression of the swelling with restoration of function and resolution of lung infiltrates. Conclusion In case of a suspicious and/or progressive swelling a definite diagnosis should be achieved by biopsy within a short time to enable a proper treatment. If scar sarcoidosis is proven further investigation is necessary to exclude a systemical involvement. A surgical treatment of the swelling is not indicated.

  1. Scar sarcoidosis on a finger mimicking a rapidly growing soft tissue tumour: a case report.

    Science.gov (United States)

    Henrichs, Marcel-Philipp; Streitbürger, Arne; Gosheger, Georg; Surke, Carsten; Dierkes, Christian; Hardes, Jendrik

    2012-10-02

    Scar sarcoidosis is a rare and uncommon but specific cutaneous manifestation of sarcoidosis. In general it arises in pre-existing scars deriving from mechanical traumas. As most surgeons dealing with scars might not be aware of cutaneous sarcoidosis and its different types of appearance the appropriate staging and treatment might be missed or at least delayed. To our knowledge this is the first case in literature of scar sarcoidosis on a finger. We present a case of a 33-year-old carpenter who developed scar sarcoidosis on his right index finger 4 years after the tendon of the long digital flexor got accidentally cut by an angle grinder. He was referred due to a swelling of the finger suspected to be a malignant soft tissue tumour. The circumference of the affected finger had almost doubled, adding up to 94 mm. Incision biopsy revealed typical noncaseating granulomas. Further investigation showed a systemic extent of the disease with involvement of the lung. A systemic treatment with oral steroids led to an almost full regression of the swelling with restoration of function and resolution of lung infiltrates. In case of a suspicious and/or progressive swelling a definite diagnosis should be achieved by biopsy within a short time to enable a proper treatment. If scar sarcoidosis is proven further investigation is necessary to exclude a systemical involvement. A surgical treatment of the swelling is not indicated.

  2. PET/MR - a rapidly growing technique of imaging in oncology and neurology.

    Science.gov (United States)

    Sałyga, Alicja; Guzikowska-Ruszkowska, Izabela; Czepczyński, Rafał; Ruchała, Marek

    2016-01-01

    The combination of positron emission tomography (PET) and magnetic resonance (MR) has become a subject of interest for researchers in the recent several years. Positron emission tomography in combination with magnetic resonance (PET/MR) is the most recent imaging technique classified in the so called hybrid systems category. This review briefly discusses the development history of PET/MR scanners, the principle of their operation, of tandem systems, as well as fully integrated devices. Further, it summarizes recent reports on the application of PET/MR scans and their possible future role in oncological and non-oncological diagnostics. Recent reports regarding the application of PET/MR scanners show huge potential of simultaneously received images, which exceed the advantages of either of those scans used separately. However, the results so far remain uncertain and require further investigations, especially in terms of clinical studies, not only for scientific purposes.

  3. A rapidly growing electronic publishing trend: audiobooks for leisure and education

    OpenAIRE

    Engelen, J J

    2008-01-01

    This contribution focuses on the relatively new phenomenon of the purely commercial availability of audiobooks, sometimes also called “spoken books”, “talking books” or “narrated books”. Having the text of a book read aloud and recorded has been for a very long time the favourite solution to make books and other texts accessible for persons with a serious reading impairment such as blindness or low vision. Specialised production centres do exist in most countries of the world for producing th...

  4. Isolation of rapid growing mycobacteria from soil and water in Iran ...

    African Journals Online (AJOL)

    A total of 350 soil samples were collected from different part of Uremia city and suburbs. We used 3% sodium lauryl sulfate and 1% NaOH for decontamination of soil samples. Of 350 samples, mycobacteria were isolated from 65 (18.3%) specimens. Mycobacterium fortuitum with 18(5.14) strains yielded the highest ...

  5. 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.

  6. Isolation of rapid growing mycobacteria from soil and water in Iran

    African Journals Online (AJOL)

    USER

    2010-06-14

    Jun 14, 2010 ... A total of 350 soil samples were collected from different part of Uremia city and suburbs. We used 3% sodium lauryl sulfate and 1% NaOH for decontamination of soil samples. Of 350 samples, mycobacteria were isolated from 65 (18.3%) specimens. Mycobacterium fortuitum with 18(5.14) strains yielded the.

  7. Mycobacterium aquaticum sp. nov., a rapidly growing species isolated from haemodialysis water.

    Science.gov (United States)

    Hashemi Shahraki, Abdolrazagh; Trovato, Alberto; Droz, Sara; Haidarieh, Parvin; Borroni, Emanuele; Mirsaeidi, Mehdi; Mannino, Roberta; Hashemzadeh, Mohamad; Mariottini, Alessandro; Cirillo, Daniela Maria; Tortoli, Enrico

    2017-09-01

    The characterization of five Iranian isolates, four from hospital haemodialysis water and one from the sputum of a patient, led to the detection of a novel mycobacterium species. The strains were characterized by mucoid colonies developing in 3-5 days at temperatures ranging from 25 to 37 °C. The biochemical test pattern was unremarkable while the HPLC profile of mycolic acids resembled that of Mycobacterium fortuitum. The sequences of three major housekeeping genes (16S rRNA, hsp65 and rpoB) were unique and differed from those of any other mycobacterium. Mycobacterium brisbanense, which is the species that shared the highest 16S rRNA gene sequence similarity (99.03 %), was distinct, as shown by the average nucleotide identity and by the genome to genome distance values (91.05 and 43.10 %, respectively). The strains are thus considered to represent a novel species of the genus Mycobacterium, for which the name Mycobacterium aquaticum sp. nov. is proposed. The type strain is RW6T (=DSM 104277T=CIP111198T).

  8. Predictors of Postpartum Depression in Dubai, a Rapidly Growing Multicultural Society in the United Arab Emirates.

    Science.gov (United States)

    Alhammadi, Salwa M; Hashem, Lien Abou; Abusbeih, Zainah R; Alzaabi, Fatima S; Alnuaimi, Salama N; Jalabi, Ala F; Nair, Satish C; Carrick, Frederick R; Abdulrahman, Mahera

    2017-09-01

    Postpartum depression (PPD) is a significant public health problem adversely affecting mothers, their newborns, and other members of the family. Although PPD is common and potentially dangerous, only a minority of the cases are identified in primary health care settings during routine care, and the majority of depressed mothers in the community lies unrecognized and therefore untreated. In this study, a total of 1500 mothers were approached randomly, 808 accepted to participate, and 504 were within the inclusion criteria (women who had a birth of a singleton full-term healthy infant, had an uncomplicated pregnancy, and were within their one week to six months postpartum). The participants completed the Edinburgh Postnatal Depression Scale. A total of 168 women had an EPDS score ≥10, yielding a crude prevalence rate of 33%. The prevalence of suicidal ideation was 14 out of 504 (3%), among which 11 (79%) had EPDS score of ≥10. We fitted multiple linear regression models to evaluate the predictors of variables measured on the EPDS scale. This model was statistically significant ppredictors. The findings of this study are anticipated to entail the government and policy makers in the region to pay more attention to the apparently high prevalence of unrevealed PPD in the community. It is crucial to enhance screening mechanisms for early detection, providing interventions to manage symptoms, and at the same time mandating local guidelines to address the PPD pathology as a high priority for the UAE population.

  9. Metal-organic frameworks: a rapidly growing class of versatile nanoporous materials.

    Science.gov (United States)

    Meek, Scott T; Greathouse, Jeffery A; Allendorf, Mark D

    2011-01-11

    Metal-organic frameworks (MOFs) represent a new class of hybrid organic-inorganic supramolecular materials comprised of ordered networks formed from organic electron donor linkers and metal cations. They can exhibit extremely high surface areas, as well as tunable pore size and functionality, and can act as hosts for a variety of guest molecules. Since their discovery, MOFs have enjoyed extensive exploration, with applications ranging from gas storage to drug delivery to sensing. This review covers advances in the MOF field from the past three years, focusing on applications, including gas separation, catalysis, drug delivery, optical and electronic applications, and sensing. We also summarize recent work on methods for MOF synthesis and computational modeling.

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

    African Journals Online (AJOL)

    rate is attributed to the influx rate, campaign design and implementation, and factors related to child and career mobi- lity. Alternative immunisation strategies, with social awareness playing a key role, are being urgently investigated. S Air Med J 1989; 76: 157-159. Measles became a notifiable disease in the RSA in 1979.1.

  11. Contralateral trigeminal neuralgia in a rapidly growing vestibular schwannoma: A case report.

    Science.gov (United States)

    Seo, Youngbeom; Kim, Dong Gyu

    2018-01-01

    We present a rare case of vestibular schwannoma (VS) with contralateral trigeminal neuralgia. A 59 year-old woman presented with right-sided trigeminal neuralgia and a small VS in the left cerebellopontine angle (CPA) that was found via magnetic resonance imaging. The patient was administered medication; however, her symptoms gradually aggravated, and the size of the tumor increased. Two years after the initial diagnosis, the patient underwent surgical resection. After surgical resection, the patient's facial pain was improved. The trigeminal neuralgia appeared to be related to brainstem displacement caused by the contralateral VS. For VS with contralateral trigeminal neuralgia, surgical resection of the tumor should be considered as a possible treatment strategy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Local circulating clones of Staphylococcus aureus in Ecuador.

    Science.gov (United States)

    Zurita, Jeannete; Barba, Pedro; Ortega-Paredes, David; Mora, Marcelo; Rivadeneira, Sebastián

    The spread of pandemic Staphylococcus aureus clones, mainly methicillin-resistant S. aureus (MRSA), must be kept under surveillance to assemble an accurate, local epidemiological analysis. In Ecuador, the prevalence of the USA300 Latin American variant clone (USA300-LV) is well known; however, there is little information about other circulating clones. The aim of this work was to identify the sequence types (ST) using a Multiple-Locus Variable number tandem repeat Analysis 14-locus genotyping approach. We analyzed 132 S. aureus strains that were recovered from 2005 to 2013 and isolated in several clinical settings in Quito, Ecuador. MRSA isolates composed 46.97% (62/132) of the study population. Within MRSA, 37 isolates were related to the USA300-LV clone (ST8-MRSA-IV, Panton-Valentine Leukocidin [PVL] +) and 10 were related to the Brazilian clone (ST239-MRSA-III, PVL-). Additionally, two isolates (ST5-MRSA-II, PVL-) were related to the New York/Japan clone. One isolate was related to the Pediatric clone (ST5-MRSA-IV, PVL-), one isolate (ST45-MRSA-II, PVL-) was related to the USA600 clone, and one (ST22-MRSA-IV, PVL-) was related to the epidemic UK-EMRSA-15 clone. Moreover, the most prevalent MSSA sequence types were ST8 (11 isolates), ST45 (8 isolates), ST30 (8 isolates), ST5 (7 isolates) and ST22 (6 isolates). Additionally, we found one isolate that was related to the livestock associated S. aureus clone ST398. We conclude that in addition to the high prevalence of clone LV-ST8-MRSA-IV, other epidemic clones are circulating in Quito, such as the Brazilian, Pediatric and New York/Japan clones. The USA600 and UK-EMRSA-15 clones, which were not previously described in Ecuador, were also found. Moreover, we found evidence of the presence of the livestock associated clone ST398 in a hospital environment. Copyright © 2016 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  13. Genotype-environment interaction and phenotypic stability for girth growth and rubber yield of Hevea clones in São Paulo State, Brazil

    Directory of Open Access Journals (Sweden)

    Gonçalves Paulo de Souza

    2003-01-01

    Full Text Available The best-yielding, best vigour and most stable Hevea clones are identified by growing clones in different environments. However, research on the stability in Hevea brasiliensis (Willd. Adr. ex Juss. Muell.-Arg. is scarce. The objectives of this work were to assess genotype-environment interaction and determine stable genotypes. Stability analysis were performed on results for girth growth and rubber yield of seven clones from five comparative trials conducted over 10 years (girth growth and four years (rubber yield in São Paulo State, Brazil. Stability was estimated using the Eberhart and Russell (1966 method. Year by location and location variability were the dominant sources of interactions. The stability analysis identified GT 1 and IAN 873 as the most stable clones for girth growth and rubber yield respectively since their regression coefficients were almost the unity (b = 1 and they had one of the lowest deviations from regressions (S2di. Their coefficient of determination (R² was as high as 89.5% and 89.8% confirming their stability. In contrast, clones such as PB 235, PR 261, and RRIM 701 for girth growth and clones such as GT 1 for rubber yield with regression coefficients greater than one were regarded as sensitive to environment changes.

  14. A simple, flexible and efficient PCR-fusion/Gateway cloning procedure for gene fusion, site-directed mutagenesis, short sequence insertion and domain deletions and swaps

    Directory of Open Access Journals (Sweden)

    Etchells J Peter

    2009-10-01

    Full Text Available Abstract Background The progress and completion of various plant genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating binary vectors containing: gene fusions, variants from site-directed mutagenesis, addition of protein tags together with domain swaps and deletions. Furthermore, efficient cloning procedures, ideally high throughput, are essential for pyramiding of multiple gene constructs. Results Here, we present a simple, flexible and efficient PCR-fusion/Gateway cloning procedure for construction of binary vectors for a range of gene fusions or variants with single or multiple nucleotide substitutions, short sequence insertions, domain deletions and swaps. Results from selected applications of the procedure which include ORF fusion, introduction of Cys>Ser mutations, insertion of StrepII tag sequence and domain swaps for Arabidopsis secondary cell wall AtCesA genes are demonstrated. Conclusion The PCR-fusion/Gateway cloning procedure described provides an elegant, simple and efficient solution for a wide range of diverse and complicated cloning tasks. Through streamlined cloning of sets of gene fusions and modification variants into binary vectors for systematic functional studies of gene families, our method allows for efficient utilization of the growing sequence and expression data.

  15. A simple, flexible and efficient PCR-fusion/Gateway cloning procedure for gene fusion, site-directed mutagenesis, short sequence insertion and domain deletions and swaps

    Science.gov (United States)

    Atanassov, Ivan I; Atanassov, Ilian I; Etchells, J Peter; Turner, Simon R

    2009-01-01

    Background The progress and completion of various plant genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating binary vectors containing: gene fusions, variants from site-directed mutagenesis, addition of protein tags together with domain swaps and deletions. Furthermore, efficient cloning procedures, ideally high throughput, are essential for pyramiding of multiple gene constructs. Results Here, we present a simple, flexible and efficient PCR-fusion/Gateway cloning procedure for construction of binary vectors for a range of gene fusions or variants with single or multiple nucleotide substitutions, short sequence insertions, domain deletions and swaps. Results from selected applications of the procedure which include ORF fusion, introduction of Cys>Ser mutations, insertion of StrepII tag sequence and domain swaps for Arabidopsis secondary cell wall AtCesA genes are demonstrated. Conclusion The PCR-fusion/Gateway cloning procedure described provides an elegant, simple and efficient solution for a wide range of diverse and complicated cloning tasks. Through streamlined cloning of sets of gene fusions and modification variants into binary vectors for systematic functional studies of gene families, our method allows for efficient utilization of the growing sequence and expression data. PMID:19863796

  16. [Cloning and expression analysis of HSP70 gene from Dendrobium officinale under low temperature stress].

    Science.gov (United States)

    Li, Dong-Bin; Gao, Han-Hui; Si, Jin-Ping; Zhu, Yu-Qiu

    2013-10-01

    To investigate HSP70 gene expression from Dendrobium officinale under low temperature stress, which will provide the molecular biological foundation for breeding the low temperature resistant strain. HSP70 gene full length cDNA was cloned by rapid amplification of cDNA ends (RACE) on the basis of HSP70 gene fragment sequences, and the structure and function of HSP70 gene were deduced. The expression of HSP70 under low temperature stress was detected by RT-PCR. The full length of HSP70 gene cDNA was 2 296 bp containing a 1 944 bp open reading frame (ORF) that encoded a protein of 647 amino acids. Its amino acids sequence had typical HSP70 characteristics and high homology with other plant's HSP70. Cold stress expression analysis showed that expression of the HSP70 gene could be induced by low temperature. The HSP70 gene of D. officinale was successfully cloned and reported for the first time which proved that the expression could be induced by low temperature. The cloning of HSP70 gene provides a stable foundation for further study of D. officinale cultivation and the breeding of the cold resistance strains.

  17. A massively parallel pipeline to clone DNA variants and examine molecular phenotypes of human disease mutations.

    Directory of Open Access Journals (Sweden)

    Xiaomu Wei

    2014-12-01

    Full Text Available Understanding the functional relevance of DNA variants is essential for all exome and genome sequencing projects. However, current mutagenesis cloning protocols require Sanger sequencing, and thus are prohibitively costly and labor-intensive. We describe a massively-parallel site-directed mutagenesis approach, "Clone-seq", leveraging next-generation sequencing to rapidly and cost-effectively generate a large number of mutant alleles. Using Clone-seq, we further develop a comparative interactome-scanning pipeline integrating high-throughput GFP, yeast two-hybrid (Y2H, and mass spectrometry assays to systematically evaluate the functional impact of mutations on protein stability and interactions. We use this pipeline to show that disease mutations on protein-protein interaction interfaces are significantly more likely than those away from interfaces to disrupt corresponding interactions. We also find that mutation pairs with similar molecular phenotypes in terms of both protein stability and interactions are significantly more likely to cause the same disease than those with different molecular phenotypes, validating the in vivo biological relevance of our high-throughput GFP and Y2H assays, and indicating that both assays can be used to determine candidate disease mutations in the future. The general scheme of our experimental pipeline can be readily expanded to other types of interactome-mapping methods to comprehensively evaluate the functional relevance of all DNA variants, including those in non-coding regions.

  18. Expression cloning and production of Human Heavy Chain Only antibodies from murine transgenic plasma cells

    Directory of Open Access Journals (Sweden)

    Dubravka Drabek

    2016-12-01

    Full Text Available Several technologies have been developed to isolate human antibodies against different target antigens as a source of potential therapeutics, including hybridoma technology, phage and yeast display systems. For conventional antibodies this involves either random pairing of VH and VL domains in combinatorial display libraries, or isolation of cognate pairs of VH and VL domains from human B cells or from transgenic mice carrying human immunoglobulin loci followed by single cell sorting, single cell RT-PCR and bulk cloning of isolated natural VH-VL pairs. Heavy chain only antibodies (HCAbs that naturally occur in camelids require only heavy immunoglobulin chain cloning. Here, we present an automatable novel, high-throughput technology, for rapid direct cloning and production of fully human HCAbs from sorted population of transgenic mouse plasma cells carrying a human HCAb locus. Utility of the technique is demonstrated by isolation of diverse sets of sequence unique, soluble, high affinity influenza A strain X-31 hemagglutinin (HA specific HCAbs

  19. Molecular cloning and characterization of Saccharomyces cerevisiae RAD28, the yeast homolog of the human Cockayne syndrome A (CSA) gene.

    OpenAIRE

    P.K Bhatia; Verhage, R.A.; Brouwer, J; Friedberg, E C

    1996-01-01

    Cockayne syndrome patients exhibit severe developmental and neurological abnormalities. Cells derived from these patients are sensitive to killing by UV radiation and do not support the rapid repair of the transcribed strand of transcriptionally active genes observed in cells from normal individuals. We report the cloning of the Saccharomyces cerevisiae homolog of the Cockayne syndrome A (CSA) gene, which we designate as RAD28. A rad28 null mutant does not manifest increased sensitivity to ki...

  20. Automated Cell-Cutting for Cell Cloning

    Science.gov (United States)

    Ichikawa, Akihiko; Tanikawa, Tamio; Matsukawa, Kazutsugu; Takahashi, Seiya; Ohba, Kohtaro

    We develop an automated cell-cutting technique for cell cloning. Animal cells softened by the cytochalasin treatment are injected into a microfluidic chip. The microfluidic chip contains two orthogonal channels: one microchannel is wide, used to transport cells, and generates the cutting flow; the other is thin and used for aspiration, fixing, and stretching of the cell. The injected cell is aspirated and stretched in the thin microchannel. Simultaneously, the volumes of the cell before and after aspiration are calculated; the volumes are used to calculate the fluid flow required to aspirate half the volume of the cell into the thin microchannel. Finally, we apply a high-speed flow in the orthogonal microchannel to bisect the cell. This paper reports the cutting process, the cutting system, and the results of the experiment.

  1. Cloned images and the optical unconscious

    DEFF Research Database (Denmark)

    Romic, Bojana

    In this presentation I would discuss the term optical unconscious as defined in the writings of Walter Benjamin and Rosalind Krauss, and propose a broadened definition of this term, taking into account the new media spreadability (Jenkins et al., 2013) and 'image cloning' (W.J.T. Mitchell, 2008...... contribute its special status. When Walter Benjamin coined the term optical unconscious, he was primarily referring to the psychoanalytical perspective: photography opened a new realm of experience that was not accessible to the naked eye – the same way that psychoanalysis provided an access to the physic...... unconscious. The camera conveys the virtuality of vision, through which the eye learns about the spatio-temporal arrangements of the photographic object 'with its devices of slow motion and enlargement' (Benjamin, 1931, pp. 510). In reference to this, Rosalind Krauss wrote in her book Optical unconscious...

  2. Avaliação de clones de banana Cavendish Evaluation of cavendish banana clones

    Directory of Open Access Journals (Sweden)

    Sebastião de Oliveira e Silva

    2006-10-01

    Full Text Available Na bananeira ocorrem variações somaclonais em taxa muito superior ao que se observa na maioria das outras culturas, provavelmente em função da instabilidade mitótica. Objetivou-se com o presente trabalho avaliar clones de bananeira Cavendish coletados em diferentes locais. Os clones Grande Naine (G.N. Taperão, G.N. Rossete, G.N. Williams, G.N. Magário, G.N. SC-074 e Nanicão (N. IAC Abóbora Verde, N. Rossete, N. SC-0008 e N. SC-063 coletados em São Paulo, Santa Catarina e Bahia foram avaliados no Lote 54-P da Thelo Produção Agropecuária (Grupo Plena, no Projeto Jaíba, Etapa 1, no município de Matias Cardoso-MG. Avaliaram-se os caracteres altura da planta, circunferência do pseudocaule, número de folhas vivas no florescimento e na colheita, número de brotos, peso do cacho e das pencas, número de frutos e de pencas por cacho, comprimento e circunferência do fruto e número de dias do plantio ao florescimento e à colheita. Observou-se grande similaridade nas características dos clones. No entanto, os resultados obtidos permitem a recomendação dos clones N. IAC Abóbora Verde e G.N. Williams.Somaclonal variations occur in bananas at greater rates compared to other crops, probably due to mitotic instability. The objective of the present research was to evaluate Cavendish banana clones collected from different sites. The 'Grand Naine' clones (G.N. Taperão, G.N. Rossete, G.N. Williams, G.N. Magário, G.N. SC-074 and 'Nanicão' (N. IAC Abóbora Verde, N. Rossete, N. SC-0008 and N. SC-063 collected from the states of São Paulo, Santa Catarina and Bahia, were evaluated at the Station 54-P of the Thelo Agricultural Production (Plena Group, in the Jaíba Project, Stage 1, in the city of Matias Cardoso-MG. The characteristics of the plant as height and pseudostem circumference, number of live leaves at flowering and harvesting, number of shoots, weight of bunch and hand, number of fruits and hands per bunch, fruit length and

  3. Cloning, recombinant expression and characterization of a new ...

    African Journals Online (AJOL)

    A new amylase gene APGA1 was cloned from Aureobasidium pullulans NRRL 12974 and expressed in Pichia pastoris. This is the first report on cloning and expression of amylolytic gene from the industrially important microorganism A. pullulans. The purified recombinant protein with MW of 66 kDa and specific activity of ...

  4. Therapeutic and reproductive cloning--implications and recommendations.

    Science.gov (United States)

    Ng, S C; Ng, Y; Liow, S L; Chen, N

    2003-03-01

    The announcement of Dolly's birth took the world by storm, mainly because what was thought impossible has become possible. Optimism that new approaches in agriculture and medicine abound, as much as fear and imagination leading to Frankenstein-like scenarios. Scientifically, cloning refers to replicating an animal with the same nuclear genetic material; whilst it may refer to embryos as the source material, the current storm refers to differentiated ("somatic") cells. Cloning technology is useful in the following areas: agriculture, to produce animals of superior or specific qualities; endangered animals, to increase genetic diversity through widening the gene pool; understanding fundamental questions in developmental embryology, through the use of laboratory animals; and in human therapy, to produce cells and possibly tissues for repair and regeneration. In the first 2 instances, it is reproductive cloning. In the last instance, it is therapeutic cloning, as no individuals are "produced". Human reproductive cloning is not allowed by all governments that have deliberated on it, and therapeutic cloning is allowed by some under certain circumstances. As therapeutic cloning has great potential in cures of many diseases, it should be allowed but with safeguards to prevent abuse and reproductive cloning in the human.

  5. Cloning and differential expression of 1- aminocyclopropane-1 ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-04-12

    Apr 12, 2010 ... Cloning of the DenACS from Dendrobium hybrid cultivar Anna was performed by RT-PCR and ..... The ACS gene of the Dendrobium hybrid cultivar Anna ..... Cloning of a cDNA encoding 1-aminocyclopropane-1- carboxylate synthase and expression of its mRNA in ripening apple fruit. Planta. 185: 38-45.

  6. Molecular cloning, expression analysis and sequence prediction of ...

    African Journals Online (AJOL)

    ajl yemi

    2011-11-28

    Nov 28, 2011 ... CCAAT/enhancer-binding protein beta as an essential transcriptional factor, regulates the differentiation of adipocytes and the deposition of fat. Herein, we cloned the whole open reading frame. (ORF) of bovine C/EBPβ gene and analyzed its putative protein structures via DNA cloning and sequence ...

  7. Evaluation of flooding tolerance in cuttings of Populus clones used ...

    African Journals Online (AJOL)

    We analysed the responses to flooding of 14 poplar clones used for forestation at the Paraná River Delta, Argentina. Some are commercial clones planted in the area, and others belong to a poplar breeding program from the National Institute of Agricultural Technology (INTA) in Argentina. Potted plants of 60 cm high ...

  8. Cloning, sequencing and expression of cDNA encoding growth ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Biosciences; Volume 26; Issue 3. Cloning, sequencing ... The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5′ and 3′ ...

  9. A set of BAC clones spanning the human genome.

    NARCIS (Netherlands)

    Krzywinski, M.; Bosdet, I.; Smailus, D.; Chiu, R.; Mathewson, C.; Wye, N.; Barber, S.; Brown-John, M.; Chan, S.; Chand, S.; Cloutier, A.; Girn, N.; Lee, D.; Masson, A.; Mayo, M.; Olson, T.; Pandoh, P.; Prabhu, A.L.; Schoenmakers, E.F.P.M.; Tsai, M.Y.; Albertson, D.; Lam, W.W.; Choy, C.O.; Osoegawa, K.; Zhao, S.; Jong, P.J. de; Schein, J.; Jones, S.; Marra, M.A.

    2004-01-01

    Using the human bacterial artificial chromosome (BAC) fingerprint-based physical map, genome sequence assembly and BAC end sequences, we have generated a fingerprint-validated set of 32 855 BAC clones spanning the human genome. The clone set provides coverage for at least 98% of the human

  10. MEANS AND METHODS FOR CLONING NUCLEIC ACID SEQUENCES

    NARCIS (Netherlands)

    Geertsma, Eric Robin; Poolman, Berend

    2008-01-01

    The invention provides means and methods for efficiently cloning nucleic acid sequences of interest in micro-organisms that are less amenable to conventional nucleic acid manipulations, as compared to, for instance, E.coli. The present invention enables high-throughput cloning (and, preferably,

  11. Cloning and transformation of SCMV CP gene and regeneration of ...

    African Journals Online (AJOL)

    The coated protein gene of sugarcane mosaic virus ( SCMV CP gene) was cloned from maize (Zea mays L.) leaves showing dwarf mosaic symptoms by reverse-transcription polymerase chain reaction (RTPCR) with degraded primers. The results of sequencing and homologous comparison indicated that the cloned gene ...

  12. Cloning and characterization of cDNA encoding xyloglucan ...

    African Journals Online (AJOL)

    We cloned complete cDNA of an XET from pearl millet (Pennisetum glaucum L.) and characterized it using in silico comparative genomics and activity assays. The cloned cDNA was 1266 bp in length, encoding a protein with 291 amino acids having signal peptide targeting it to the cell wall. The protein showed xyloglucan ...

  13. Cloning and expression of Toxoplasma gondii tachyzoite P22 protein

    African Journals Online (AJOL)

    Recombinant plasmid was transformed in E. coli (Bl21 DE3) and induced by 1 mM IPTG and analyzed by 12% SDS-PAGE. Expressd protein was purified by affinity chromatography and confirmed by western blot analysis. We successfully cloned and expressed T. gondii P22 protein. Key words: Toxoplasma gondii, cloning, ...

  14. Cloning and characterization of promoter and 5 '-UTR of the NMDA receptor subunit epsilon(2) : evidence for alternative splicing of 5 '-non-coding exon

    NARCIS (Netherlands)

    Klein, M; Pieri, [No Value; Uhlmann, F; Pfizenmaier, K; Eisel, U

    1998-01-01

    Using rapid amplification of cDNA ends (RACE), we have cloned the 5'-untranslated region (5'-UTR) of the N-methyl-D-aspartate receptor subunit epsilon(2) from murine forebrain-derived mRNA. We identified two distinct types of cDNA species differing in the presence or absence of one exon sequence.

  15. Cloning and characterization of promoter and 5'-UTR of the NMDA receptor subunit ε2 : evidence for alternative splicing of 5'-non-coding exon

    NARCIS (Netherlands)

    Klein, Matthias; Pieri, Isabelle; Uhlmann, Frank; Pfizenmaier, Klaus; Eisel, Ulrich

    1998-01-01

    Using rapid amplification of cDNA ends (RACE), we have cloned the 5'-untranslated region (5'-UTR) of the N-methyl-D-aspartate receptor subunit ε2 from murine forebrain-derived mRNA. We identified two distinct types of cDNA species differing in the presence or absence of one exon sequence. Sequencing

  16. 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.

  17. Growing pioneer plants for a lunar base

    Science.gov (United States)

    Kozyrovska, N. O.; Lutvynenko, T. L.; Korniichuk, O. S.; Kovalchuk, M. V.; Voznyuk, T. M.; Kononuchenko, O.; Zaetz, I.; Rogutskyy, I. S.; Mytrokhyn, O. V.; Mashkovska, S. P.; Foing, B. H.; Kordyum, V. A.

    A precursory scenario of cultivating the first plants in a lunar greenhouse was elaborated in frames of a conceptual study to grow plants for a permanently manned lunar base. A prototype plant growth system represents an ornamental plant Tagetes patula L. for growing in a lunar rock anorthosite as a substrate. Microbial community anticipated to be in use to support a growth and development of the plant in a substrate of low bioavailability and provide an acceptable growth and blossoming of T. patula under growth limiting conditions.

  18. Using adult cloned trees grown under natural conditions to characterize BVOC emission variation

    Science.gov (United States)

    Persson, Ylva; Schurgers, Guy; Ekberg, Anna; Rinnan, Riikka; Holst, Thomas

    2015-04-01

    Biogenic Volatile Organic Compounds (BVOCs) are diverse chemical species produced and emitted from the vegetation as trace gases. BVOCs are commonly grouped into isoprene, monoterpenes and sesquiterpenes, where isoprene is mainly emitted by deciduous trees and monoterpenes and sesquiterpenes by coniferous trees. BVOCs are known to have a considerable impact on atmospheric chemistry and are precursors for secondary organic aerosol, which in turn are important for the aerosol feedback on the Earth's climate. Recently, Bäck et al. (2012) reported a high diversity of the chemical composition of emitted compounds from pine trees growing at the same stand due to genetic variation. This study here uses cloned trees growing naturally in a transect in Europe in order to exclude genetic variation and to assess emission variation between and within selected tree species grown at different climatic conditions. The International Phenological Garden (IPG) network, where cloned trees are used to monitor the long-term phenological observations of representative tree species for Europe provides a specific, cloned set of important tree species, which had been planted throughout Europe starting in 1957. This gives a unique opportunity to study the adaptation to various climatic conditions and field conditions in genetically identical plants in relation to BVOC emissions. During a field campaign in 2013 at the IPG site in Taastrup, Denmark (55°40' N, 12°18' E), seven trees were measured at three heights within the canopy. Measured trees were two English oaks (Quercus robur), one European beech (Fagus sylvatica) and four Norway spruces (Picea abies) of two provenances. For oak and one provenance of spruce, measurements were performed twice, both in June and in August in order to examine any emission pattern change with the progression of the summer. Measurements were performed using a gas-exchange cuvette of a photosynthesis system combined with BVOC adsorbent tubes, which were

  19. USER-derived cloning methods and their primer design.

    Science.gov (United States)

    Salomonsen, Bo; Mortensen, Uffe H; Halkier, Barbara A

    2014-01-01

    Uracil excision-based cloning through USER™ (Uracil-Specific Excision Reagent) is an efficient ligase-free cloning technique that comprises USER cloning, USER fusion, and USER cassette-free (UCF) USER fusion. These USER-derived cloning techniques enable seamless assembly of multiple DNA fragments in one construct. Though governed by a few simple rules primer design for USER-based fusion of PCR fragments can prove time-consuming for inexperienced users. The Primer Help for USER (PHUSER) software is an easy-to-use primer design tool for USER-based methods. In this chapter, we present a PHUSER software protocol for designing primers for USER-derived cloning techniques.

  20. pPCV, a versatile vector for cloning PCR products.

    Science.gov (United States)

    Janner, Christiane R; Brito, Ana Lívia P; Moraes, Lidia Maria P; Reis, Viviane Cb; Torres, Fernando Ag

    2013-01-01

    The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3' adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3' overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that purpose, a minimal polylinker was designed to include restriction sites for EcoRV and XcmI which enable direct cloning of amplicons bearing blunt-ends or A-overhangs, respectively, still offering blue/white selection. When tested, the resulting vector, pPCV, presented high efficiency cloning of both types of amplicons.

  1. [TSA improve transgenic porcine cloned embryo development and transgene expression].

    Science.gov (United States)

    Kong, Qing-Ran; Zhu, Jiang; Huang, Bo; Huan, Yan-Jun; Wang, Feng; Shi, Yong-Qian; Liu, Zhong-Feng; Wu, Mei-Ling; Liu, Zhong-Hua

    2011-07-01

    Uncompleted epigenetic reprogramming is attributed to the low efficiency of producing transgenic cloned animals. Histone modification associated with epigenetics can directly influence the embryo development and transgene expression. Trichostatin A (TSA), as an inhibitor of histone deacetylase, can change the status of histone acetylation, improve somatic cell reprogramming, and enhance cloning efficiency. TSA prevents the chromatin structure from being condensed, so that transcription factor could binds to DNA sequence easily and enhance transgene expression. Our study established the optimal TSA treatment on porcine donor cells and cloned embryos, 250 nmol/L, 24 h and 40 nmol/L, 24 h, respectively. Furthermore, we found that both the cloned embryo and the donor cell treated by TSA resulted in the highest development efficiency. Meanwhile, TSA can improve transgene expression in donor cell and cloned embryo. In summary, TSA can significantly improve porcine reconstructed embryo development and transgene expression.

  2. The science and technology of farm animal cloning

    DEFF Research Database (Denmark)

    Gjerris, Mickey; Vajta, Gábor

    . The major breakthrough in terms of public recognition has happened when Ian Wilmut et al. [Wilmut, I., Schnieke, A.E., McWhir, J., Kind, A.J., Campbell, K.H., 1997. Viable offspring derived from fetal és adult mammalian cells. Nature 385, 810-813] described the successful application of almost exactly......, goats, horses, cats, etc. have been cloned with the somatic cell nuclear transfer technique. Although the technology still has relatively low success rates and there seems to be substantial problems with the welfare of some of the cloned animals, cloning is used both within basic research...... and the biomedical sector. The next step seems to be to implement cloning in the agricultural production system and several animals have been developed in this direction. This article reviews the current state of the art of farm animal cloning from a scientific and technological perspective, describes the animal...

  3. A new trilocus sequence-based multiplex-PCR to detect major Acinetobacter baumannii clones.

    Science.gov (United States)

    Martins, Natacha; Picão, Renata Cristina; Cerqueira-Alves, Morgana; Uehara, Aline; Barbosa, Lívia Carvalho; Riley, Lee W; Moreira, Beatriz Meurer

    2016-08-01

    A collection of 163 Acinetobacter baumannii isolates detected in a large Brazilian hospital, was potentially related with the dissemination of four clonal complexes (CC): 113/79, 103/15, 109/1 and 110/25, defined by University of Oxford/Institut Pasteur multilocus sequence typing (MLST) schemes. The urge of a simple multiplex-PCR scheme to specify these clones has motivated the present study. The established trilocus sequence-based typing (3LST, for ompA, csuE and blaOXA-51-like genes) multiplex-PCR rapidly identifies international clones I (CC109/1), II (CC118/2) and III (CC187/3). Thus, the system detects only one (CC109/1) out of four main CC in Brazil. We aimed to develop an alternative multiplex-PCR scheme to detect these clones, known to be present additionally in Africa, Asia, Europe, USA and South America. MLST, performed in the present study to complement typing our whole collection of isolates, confirmed that all isolates belonged to the same four CC detected previously. When typed by 3LST-based multiplex-PCR, only 12% of the 163 isolates were classified into groups. By comparative sequence analysis of ompA, csuE and blaOXA-51-like genes, a set of eight primers was designed for an alternative multiplex-PCR to distinguish the five CC 113/79, 103/15, 109/1, 110/25 and 118/2. Study isolates and one CC118/2 isolate were blind-tested with the new alternative PCR scheme; all were correctly clustered in groups of the corresponding CC. The new multiplex-PCR, with the advantage of fitting in a single reaction, detects five leading A. baumannii clones and could help preventing the spread in healthcare settings. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Rapid Prototyping Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — The ARDEC Rapid Prototyping (RP) Laboratory was established in December 1992 to provide low cost RP capabilities to the ARDEC engineering community. The Stratasys,...

  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. Reproductive cloning in humans and therapeutic cloning in primates: is the ethical debate catching up with the recent scientific advances?

    Science.gov (United States)

    Camporesi, S; Bortolotti, L

    2008-09-01

    After years of failure, in November 2007 primate embryonic stem cells were derived by somatic cellular nuclear transfer, also known as therapeutic cloning. The first embryo transfer for human reproductive cloning purposes was also attempted in 2006, albeit with negative results. These two events force us to think carefully about the possibility of human cloning which is now much closer to becoming a reality. In this paper we tackle this issue from two sides, first summarising what scientists have achieved so far, then discussing some of the ethical arguments in favour and against human cloning which are debated in the context of policy making and public consultation. Therapeutic cloning as a means to improve and save lives has uncontroversial moral value. As to human reproductive cloning, we consider and assess some common objections and failing to see them as conclusive. We do recognise, though, that there will be problems at the level of policy and regulation that might either impair the implementation of human reproductive cloning or make its accessibility restricted in a way that could become difficult to justify on moral grounds. We suggest using the time still available before human reproductive cloning is attempted successfully to create policies and institutions that can offer clear directives on its legitimate applications on the basis of solid arguments, coherent moral principles, and extensive public consultation.

  7. Growing up Female: As Six become One

    Science.gov (United States)

    Balto, Karen; Balto, Sylvia

    1971-01-01

    Growing up Female: As Six Become One" is an excellent film for people of any age and appropriate for any group. The film explores the lives of six American women showing how they are socialized and what their roles in society are. (Author)

  8. Growing Languages with Metamorphic Syntax Macros

    DEFF Research Database (Denmark)

    Brabrand, Claus; Schwartzbach, Michael Ignatieff

    2002-01-01

    simultaneously on multiple parse trees at once. The result is a highly flexible mechanism for growing new language constructs without resorting to compile-time programming. In fact, whole new languages can be defined at surprisingly low cost.This work is fully implemented as part of the bigwig system...

  9. Pueblo Girls: Growing Up in Two Worlds.

    Science.gov (United States)

    Keegan, Marcia

    This book portrays San Ildefonso Pueblo on the east bank of the Rio Grande river in New Mexico through the lives of Sonja, age 10, and her sister Desiree, age 8. Growing up in San Ildefonso Pueblo, the girls enjoy the same activities as other American girls, such as basketball, cheerleading, playing video games, and sending e-mail. But they also…

  10. What a Pain! Kids and Growing Pains

    Science.gov (United States)

    ... Steven Dowshen, MD Date reviewed: June 2015 For Teens For Kids For Parents MORE ON THIS TOPIC Why Do I Have Pain? How Do Pain Relievers Work? Growth Disorders I'm Growing Up - But Am I Normal? What Medicines Are and What They Do Feeling Too Tall or Too Short Contact Us Print Resources Send ...

  11. Growing Income Inequality Threatens American Education

    Science.gov (United States)

    Duncan, Greg J.; Murnane, Richard J.

    2014-01-01

    The first of two articles in consecutive months describes the origins and nature of growing income inequality, and some of its consequences for American children. It documents the increased family income inequality that's occurred over the past 40 years and shows that the increased income disparity has been more than matched by an expanding…

  12. Growing Up in Germany: A National Report.

    Science.gov (United States)

    Krappmann, Lothar

    1999-01-01

    Summarizes a Federal Ministry of Youth report on the conditions under which children grow up in Germany. Notes manifold problems that children face under today's living conditions. Presents recommendations and suggestions for providing a network of measures, relationships, and institutions to support children's development and education in family,…

  13. Determination of responses of growing pigs to

    African Journals Online (AJOL)

    1998-03-09

    Mar 9, 1998 ... The responses in growing pigs to balanced diets at different dietary energy levels are estimated from published data after recalculation of digestible energy (DE) iev- els using standard tables. Although responses in live weight gain (ADG), food intake (Fl), digestible energy intake (DEI) and food conversion ...

  14. How fast to northern hardwoods grow?

    Science.gov (United States)

    Rapheal Zon; H.F. Scholz

    1929-01-01

    The knowledge of the rate at which trees grow in virgin forests, after clear cutting and under selective logging, is indispensable in any forest calculations or forest practice. The enactment of the Forest Crop Law, which brought under its operation about 175,000 acres of cut-over land during the first year, the example set by several progressive lumbermen in selective...

  15. Growing Up in an Alcoholic Family.

    Science.gov (United States)

    Abbott, Stephanie

    1993-01-01

    Discusses problems faced by children growing up in an alcoholic family. Reviews four survivor roles of children of alcoholics (COAs): super-coper, scapegoat, lost child, and family mascot. Describes alcoholism as a disease of denial. Reviews the Children of Alcoholics movement begun by adult COAs to become advocates for COAs. (NB)

  16. Growing up as a Young Artist

    Science.gov (United States)

    Szekely, George

    2012-01-01

    "Growing Up as a Young Artist" is an illustrated book assignment that involves researching family scrapbooks, photo albums and films, and inquiring about family anecdotes for clues to one's artistic roots. Students creatively reflect on their early memories of imaginative events, as each page is filled with memories of creative activities they…

  17. Play Games to Grow up Bilingual

    DEFF Research Database (Denmark)

    Valente, Andrea; Marchetti, Emanuela

    2007-01-01

    A new kind of computer game is proposed, to support the linguistic development of primary school children, growing in multilingual environments: with it players will be able to simultaneously learn multiple languages. The novel idea is to treat words in different languages as physical items, that...

  18. Play Games to Grow up Bilingual

    DEFF Research Database (Denmark)

    Valente, Andrea; Marchetti, Emanuela

    2008-01-01

    A new kind of computer game is proposed, to support the linguistic development of primary school children, growing in multilingual environments: with it players will be able to simultaneously learn multiple languages. The novel idea is to treat words in different languages as physical items, that...

  19. Preliminary characterization of slow growing rhizobial strains ...

    African Journals Online (AJOL)

    In this paper, we did some preliminary characterization of six slow growing rhizobial strains, isolated from Retama monosperma (L.) Boiss. root nodules sampled from 3 sites along the coast of Oran (CapeFalcon, Bousfer and MersElHadjadj) in Northwestern Algeria. Results of this study showed that all strains had a very ...

  20. growing African giant rats Cricetomys gambianus

    African Journals Online (AJOL)

    Thermoregulation and evaporative water loss in growing African giant rats Cricetomys gambianus. M.H. Knight. Mammal Research Institute, University of Pretoria, Pretoria. With an increase in mass, weaned giant rat pups. Cricetomys gambianus, showed a corresponding decline in mass specific metabolism, conductance ...