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  • 1
    In: eLife, eLife Sciences Publications, Ltd, Vol. 12 ( 2023-07-20)
    Abstract: Fruit flies are widely used in the life and biomedical sciences as models of animal biology. They are small in size and easy to care for in a laboratory, making them ideal for studying how the body works. There are, however, some experiments that are difficult to perform on whole flies and it would be advantageous to use populations of fruit fly cells grown in the laboratory – known as cell cultures – instead. Unlike studies in humans and other mammals, which – for ethical and practical reasons –heavily rely on cell cultures, few studies have used fruit fly cell cultures. Recent work has shown that having an always active version of a gene called Ras in fruit fly cells helps the cells to survive and grow in cultures, making it simpler to generate new fruit fly cell lines compared with traditional methods. However, the methods used to express activated Ras result in cell lines that can be a mixture of many different types of cell, which limits how useful they are for research. Here, Coleman-Gosser, Hu, Raghuvanshi, Stitzinger et al. aimed to use Ras to generate a collection of cell lines from specific types of fruit fly cells in the muscle, nervous system, blood and other parts of the body. The experiments show that selectively expressing activated Ras in an individual type of cell enables them to outcompete other cells in culture to generate a cell line consisting only of the cell type of interest. The new cell lines offer models for experiments that more closely reflect their counterparts in flies. For example, the team were able to recapitulate how fly muscles develop by treating one of the cell lines with a hormone called ecdysone, which triggered the cells to mature into active muscle cells that spontaneously contract and relax. In the future, the new cell lines could be used for various experiments including high throughput genetic screening or testing the effects of new drugs and other compounds. The method used in this work may also be used by other researchers to generate more fruit fly cell lines.
    Type of Medium: Online Resource
    ISSN: 2050-084X
    Language: English
    Publisher: eLife Sciences Publications, Ltd
    Publication Date: 2023
    detail.hit.zdb_id: 2687154-3
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  • 2
    Online Resource
    Online Resource
    Wiley ; 2019
    In:  Journal of Paediatrics and Child Health Vol. 55, No. 4 ( 2019-04), p. 468-471
    In: Journal of Paediatrics and Child Health, Wiley, Vol. 55, No. 4 ( 2019-04), p. 468-471
    Type of Medium: Online Resource
    ISSN: 1034-4810 , 1440-1754
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2019
    detail.hit.zdb_id: 2007577-7
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  • 3
    In: G3 Genes|Genomes|Genetics, Oxford University Press (OUP), Vol. 12, No. 2 ( 2022-02-04)
    Abstract: Drosophila cell lines are used by researchers to investigate various cell biological phenomena. It is crucial to exercise good cell culture practice. Poor handling can lead to both inter- and intra-species cross-contamination. Prolonged culturing can lead to introduction of large- and small-scale genomic changes. These factors, therefore, make it imperative that methods to authenticate Drosophila cell lines are developed to ensure reproducibility. Mammalian cell line authentication is reliant on short tandem repeat (STR) profiling; however, the relatively low STR mutation rate in Drosophila melanogaster at the individual level is likely to preclude the value of this technique. In contrast, transposable elements (TEs) are highly polymorphic among individual flies and abundant in Drosophila cell lines. Therefore, we investigated the utility of TE insertions as markers to discriminate Drosophila cell lines derived from the same or different donor genotypes, divergent sub-lines of the same cell line, and from other insect cell lines. We developed a PCR-based next-generation sequencing protocol to cluster cell lines based on the genome-wide distribution of a limited number of diagnostic TE families. We determined the distribution of five TE families in S2R+, S2-DRSC, S2-DGRC, Kc167, ML-DmBG3-c2, mbn2, CME W1 Cl.8+, and ovarian somatic sheath Drosophila cell lines. Two independent downstream analyses of the next-generation sequencing data yielded similar clustering of these cell lines. Double-blind testing of the protocol reliably identified various Drosophila cell lines. In addition, our data indicate minimal changes with respect to the genome-wide distribution of these five TE families when cells are passaged for at least 50 times. The protocol developed can accurately identify and distinguish the numerous Drosophila cell lines available to the research community, thereby aiding reproducible Drosophila cell culture research.
    Type of Medium: Online Resource
    ISSN: 2160-1836
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2022
    detail.hit.zdb_id: 2629978-1
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  • 4
    In: BIO-PROTOCOL, Bio-Protocol, LLC, Vol. 11, No. 15 ( 2021)
    Type of Medium: Online Resource
    ISSN: 2331-8325
    Language: English
    Publisher: Bio-Protocol, LLC
    Publication Date: 2021
    detail.hit.zdb_id: 2833269-6
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  • 5
    In: Pediatric Infectious Disease Journal, Ovid Technologies (Wolters Kluwer Health), Vol. 41, No. 1 ( 2022-01), p. 1-5
    Abstract: The North Queensland region of Australia has a high incidence of pediatric thoracic empyema (pTE). We describe the management of empyema at the Townsville University Hospital which is the regional referral center for these children. The impact of a newly developed institutional guideline is also discussed. Methods: This retrospective audit included children under the age of 16 years treated for empyema between 1 Jan 2007 and 31 December 2018. Demographic and management-related variables were correlated to outcomes. A local guideline was introduced at the beginning of 2017 and patient outcomes characteristics pre, and post introduction of this guideline are compared. Results: There were 153 children with pTE (123 before and 30 after the introduction of a local guideline). Nonsurgical management was associated with a higher treatment failure rate. Median length of stay (LOS) was 11.8 (IQR 9.3–16) days. Longer hospital LOS was associated with younger age (r 2 −0.16, P = 0.04), Aboriginal and/or Torres Strait (ATSI) ancestry (13.8 vs. 10.5 days, P = 0.002) and concomitant respiratory viral infections (14.4 vs. 10.9 days, P = 0.003). The introduction of local guideline was associated with significant decrease in the use of empirical chest CT scans (54.4% before vs. 6.7% after, P 〈 0.001) and duration of intravenous antibiotics (14 days before vs. 10 days after, P = 0.02). There was no significant change in the hospital LOS (12.1 days pre and 11.7 post, P = 0.8). Conclusions: Younger age, concomitant viral respiratory infections and ATSI ancestry were identified as potential risk factors for increase LOS. Hospital LOS following the adoption of an institutional guideline was unchanged. However, such a guideline may identify populations at risk for an unfavorable course and avoid unnecessary antibiotic treatment and radiation exposure.
    Type of Medium: Online Resource
    ISSN: 0891-3668
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 2020216-7
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  • 6
    Online Resource
    Online Resource
    Oxford University Press (OUP) ; 2020
    In:  G3 Genes|Genomes|Genetics Vol. 10, No. 12 ( 2020-12-01), p. 4541-4551
    In: G3 Genes|Genomes|Genetics, Oxford University Press (OUP), Vol. 10, No. 12 ( 2020-12-01), p. 4541-4551
    Abstract: Successful Drosophila cell culture relies on media containing xenogenic components such as fetal bovine serum to support continuous cell proliferation. Here, we report a serum-free culture condition that supports the growth and proliferation of Drosophila S2R+ and Kc167 cell lines. Importantly, the gradual adaptation of S2R+ and Kc167 cells to a media lacking serum was supported by supplementing the media with adult Drosophila soluble extract, commonly known as fly extract. The utility of these adapted cells lines is largely unchanged. The adapted cells exhibited robust proliferative capacity and a transfection efficiency that was comparable to control cells cultured in serum-containing media. Transcriptomic data indicated that the S2R+ cells cultured with fly extract retain their hemocyte-specific transcriptome profile, and there were no global changes in the transcriptional output of cell signaling pathways. Our metabolome studies indicate that there were very limited metabolic changes. In fact, the cells were likely experiencing less oxidative stress when cultured in the serum-free media supplemented with fly extract. Overall, the Drosophila cell culture conditions reported here consequently provide researchers with an alternative and physiologically relevant resource to address cell biological research questions.
    Type of Medium: Online Resource
    ISSN: 2160-1836
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2020
    detail.hit.zdb_id: 2629978-1
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  • 7
    Online Resource
    Online Resource
    Oxford University Press (OUP) ; 2021
    In:  G3 Genes|Genomes|Genetics Vol. 11, No. 8 ( 2021-08-07)
    In: G3 Genes|Genomes|Genetics, Oxford University Press (OUP), Vol. 11, No. 8 ( 2021-08-07)
    Abstract: The generation of Drosophila stable cell lines has become invaluable for complementing in vivo experiments and as tools for genetic screens. Recent advances utilizing attP/PhiC31 integrase system has permitted the creation of Drosophila cells in which recombination mediated cassette exchange (RMCE) can be utilized to generate stably integrated transgenic cell lines that contain a single copy of the transgene at the desired locus. Current techniques, besides being laborious and introducing extraneous elements, are limited to a handful of cell lines of embryonic origin. Nonetheless, with well over 100 Drosophila cell lines available, including an ever-increasing number CRISPR/Cas9 modified cell lines, a more universal methodology is needed to generate a stably integrated transgenic line from any one of the available Drosophila melanogaster cell lines. Here, we describe a toolkit and procedure that combines CRISPR/Cas9 and theaaa PhiC31 integrase system. We have generated and isolated single cell clones containing an Actin5C::dsRed cassette flanked by attP sites into the genome of Kc167 and S2R+ cell lines that mimic the in vivo attP sites located at 25C6 and 99F8 of the Drosophila genome. Furthermore, we tested the functionality of the attP docking sites utilizing two independent GFP expressing constructs flanked by attB sites that permit RMCE and therefore the insertion of any DNA of interest. Lastly, to demonstrate the universality of our methodology and existing constructs, we have successfully integrated the Actin5C::dsRed cassette flanked by attP sites into two different CNS cell lines, ML-DmBG2-c2 and ML-DmBG3-c2. Overall, the reagents and methodology reported here permit the efficient generation of stable transgenic cassettes with minimal change in the cellular genomes in existing D. melanogaster cell lines.
    Type of Medium: Online Resource
    ISSN: 2160-1836
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2021
    detail.hit.zdb_id: 2629978-1
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  • 8
    Online Resource
    Online Resource
    Wiley ; 2022
    In:  ANZ Journal of Surgery Vol. 92, No. 12 ( 2022-12), p. 3293-3297
    In: ANZ Journal of Surgery, Wiley, Vol. 92, No. 12 ( 2022-12), p. 3293-3297
    Abstract: There is limited literature on renal abscesses in children and therefore no consensus on management. The objectives of this study were to describe renal abscesses in a contemporary paediatric Australian population and present a 20 year review of the literature. Methods An 11 year retrospective comparative study was conducted of paediatric patients with renal abscesses. A literature review of all eight original articles on paediatric renal abscesses from January 2001 to December 2021 was performed. Results Fourteen children with a mean age of 11 years were diagnosed with a renal abscess on ultrasound and/or computed tomography. The most common presenting symptoms were fever ( n  = 13, 93%) and flank or abdominal pain ( n  = 12, 86%). The most common causative organisms were Staphylococcus aureus ( n  = 7, 50%) and Escherichia coli ( n  = 4, 29%). All renal abscesses less than 3 cm were managed with antibiotics alone. Five out of nine abscesses 3–5 cm were managed with percutaenous drainage (56%). Two multi‐loculated abscesses greater than 5 cm required open drainage in theatre (100%). Conclusions The most common causative organism in the North Queensland population was S. aureus , with a higher incidence of MRSA. This should be taken into consideration when prescribing empirical antibiotics. Most renal abscesses in children that are less than 3 cm in size can be managed with antibiotic therapy only. The evidence for management of larger abscesses is less clear, but where clinically appropriate conservative management with antibiotic therapy should be considered in the first instance, with percutaneous drainage in cases of antibiotic failure.
    Type of Medium: Online Resource
    ISSN: 1445-1433 , 1445-2197
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2095927-8
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