Publications by authors named "Moyle P"

107 Publications

YY-11, a camel milk-derived peptide, inhibits TGF-β-mediated atherogenic signaling in human vascular smooth muscle cells.

J Food Biochem 2021 Jul 27:e13882. Epub 2021 Jul 27.

School of Pharmacy, University of Queensland, Pharmacy Australia Centre of Excellence, Woolloongabba, QLD, Australia.

Atherosclerosis, the major underlying pathology of cardiovascular disease, commences with the binding and trapping of lipids on modified proteoglycans, with hyperelongated glycosaminoglycan chains. Transforming growth factor (TGF)-β stimulates glycosaminoglycan elongation in vascular smooth muscle cells. We have recently shown that this TGF-β signaling pathway involves reactive oxygen species (ROS). YY-11 is a dodecapeptide derived from camel milk and it has antioxidant activity. We have investigated the role of YY-11 in blocking ROS signaling and downstream atherogenic responses. YY-11 inhibited TGF-β stimulated ROS production and inhibited the expression of genes for glycosaminoglycan chain elongation as a component of an in vitro model of atherosclerosis. This study provides a biochemical mechanism for the role of camel milk as a potential nutritional product to contribute to the worldwide amelioration of cardiovascular disease. PRACTICAL APPLICATIONS: The identification of readily accessible foods with antioxidant properties would provide a convenient and cost-effective approach community wide reducing oxidative stress induced pathologies such as atherosclerosis. We demonstrate that camel milk-derived peptide is an antioxidant that can inhibit growth factor-mediated proteoglycan modification in vitro. As proteoglycan modification is being recognized as one of the earliest atherogenic responses, these data support the notion of camel milk as a suitable nutritional product to contribute to the prevention of early stage of atherosclerosis development.
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http://dx.doi.org/10.1111/jfbc.13882DOI Listing
July 2021

Sebaceous carcinoma of the chest wall: A case report.

Radiol Case Rep 2021 Jul 25;16(7):1870-1873. Epub 2021 May 25.

Consultant Breast Surgeon Cambridge University Hospital NHS Trust, Cambridge and Anglia Ruskin School of Medicine, Cambridge, UK.

Sebaceous carcinoma is a rare, malignant tumor of the sebaceous glands. This is a case report of a 75-year-old man who presented with a right lower axillary mass which initially was considered to be a benign sebaceous cyst from sonography. The lesion rapidly changed in clinical and sonographic appearances and on histology was a sebaceous carcinoma. This unusual tumor accounts for less than 1% of all cutaneous malignant tumors, and the chest wall is a rare extraocular site with only nine cases described in the literature. Immunohistochemistry was negative for MSH2 and MSH6, and positive for MLH1 and PMS2, raising the possibility of Lynch or Muir-Torre Syndrome which is a known association with this rare tumor.
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http://dx.doi.org/10.1016/j.radcr.2021.04.060DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8166907PMC
July 2021

Developing GLP-1 Conjugated Self-Assembling Nanofibers Using Copper-Catalyzed Alkyne-Azide Cycloaddition and Evaluation of Their Biological Activity.

Bioconjug Chem 2021 04 12;32(4):810-820. Epub 2021 Apr 12.

School of Pharmacy, The University of Queensland, Woolloongabba 4102, Australia.

Glucagon-like peptide-1 GLP-1 is a gut-derived peptide secreted from pancreatic β-cells that reduces blood glucose levels and body weight; however, native GLP-1 (GLP-1(7-36)-NH and GLP-1(7-37)) have short circulation half-lives (∼2 min) due to proteolytic degradation and rapid renal clearance due to its low molecular weight (MW; 3297.7 Da). This study aimed to improve the proteolytic stability and delivery properties of glucagon-like peptide-1 (GLP-1) through modifications that form nanostructures. For this purpose, N- () and C-terminal (), and Lys26 side chain () alkyne-modified GLP-1 analogues were conjugated to an azide-modified lipidic peptide () to give , , and , respectively; or was conjugated with a fibrilizing self-assembling peptide (SAP) (AEAEAKAK) to yield , using copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC). demonstrated the best serum stability ( > 48 h) compared to (44 h), (20 h), (27 h), and the parental GLP-1(7-36;A8G)-NH () (19 h) peptides. Each conjugate demonstrated subnanomolar hGLP-1RA potency, and none demonstrated toxicity toward PC-3 cells at concentrations up to 1 μM. Each analogue was observed by transmission electron microscopy to form fibrils in solution. demonstrated among the best human serum stability ( = 44 h) and similar hGLP-1RA potency (EC 48 pM) to . In conclusion, this study provided an alternative to lipid modification, i.e., fibrillizing peptides, that could improve pharmacokinetic parameters of GLP-1.
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http://dx.doi.org/10.1021/acs.bioconjchem.1c00091DOI Listing
April 2021

Sortase A (SrtA) inhibitors as an alternative treatment for superbug infections.

Drug Discov Today 2021 Mar 27. Epub 2021 Mar 27.

Institute for Molecular Bioscience, The University of Queensland, St Lucia, Qld 4067, Australia. Electronic address:

Virulence factor, sortase A (SrtA), has crucial roles in the pathogenesis of Gram-positive superbugs. SrtA is a bacterial cell membrane enzyme that anchors crucial virulence factors to the cell wall surface of Gram-positive bacteria. SrtA is not necessary for bacterial growth and viability and is conveniently accessible in the cell membrane; therefore, it is an ideal target for antivirulence drug development. In this review, we focus on antimicrobial resistance (AMR)-expressing bacteria and SrtA as a potential target for overcoming AMR. The mechanism of action of SrtA and its inhibition by various types of inhibitors, such as synthetic small molecules, peptides, and natural products, are provided. Future SrtA research perspectives for alternative drug development to antibiotics are also proposed.
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http://dx.doi.org/10.1016/j.drudis.2021.03.019DOI Listing
March 2021

Introduction of an abbreviated breast MRI service in the UK as part of the BRAID trial: practicalities, challenges, and future directions.

Clin Radiol 2021 Jun 9;76(6):427-433. Epub 2021 Mar 9.

Department of Radiology, University of Cambridge School of Clinical Medicine, Box 218, Level 5, Cambridge Biomedical Campus, Cambridge, CB2 0QQ, UK. Electronic address:

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http://dx.doi.org/10.1016/j.crad.2021.01.020DOI Listing
June 2021

Recurrent myoid hamartoma of the breast mimicking malignancy.

Radiol Case Rep 2021 Feb 30;16(2):295-299. Epub 2020 Nov 30.

Cambridge University Hospitals NHS Foundation Trust, United Kingdom.

Myoid (muscular) hamartoma is a rare form of benign breast hamartoma composed of differentiated mammary glandular and stromal structures, fatty tissue and areas of smooth muscle from which its name originates. It is considered to be a variant of a mammary hamartoma. We report the clinical presentation, imaging appearances and treatment of the initial and recurrent presentation of this rare tumour in a 61year old female, which mimicked malignancy.  Although rare, myoid hamartoma's can reoccur and when they do they imaging appearances of benign and malignant tumours can overlap tend to mimic malignancy and histological diagnosis is mandatory.
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http://dx.doi.org/10.1016/j.radcr.2020.11.007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7710508PMC
February 2021

Achieving margin clearance following oncoplastic breast surgery in comparison with simple wide local excision: a three-dimensional specimen assessment.

ANZ J Surg 2020 11 17;90(11):2346-2352. Epub 2020 Aug 17.

Breast Surgery, Cambridge University Hospitals, Cambridge, UK.

Background: Pre-operative breast tumour radial dimensions often determine the choice between simple wide local excision (WLE) and oncoplastic breast surgery (OBS). We reviewed the three-dimensional interplay between tumour and surgical specimen dimensions in the two cohorts.

Methods: Demographic, tumour and treatment data were collected for all patients undergoing OBS by a single surgeon and compared with a randomly selected cohort of WLE patients treated. The relationship between tumour and specimen medio-lateral, supero-inferior and antero-posterior dimensions were explored in both groups. Subgroup analyses were performed in the OBS cohort (parenchymal displacement versus replacement).

Results: We identified 60 OBS patients (63 breasts), comparing them with 60 WLE patients. Pre-operative tumour estimated size was significantly larger in the OBS cohort and concordant with macroscopic tumour radial dimensions and final microscopic tumour size. Surgical specimen weight was more than 3.5 times higher in the OBS group and its radial dimensions were almost double. No significant difference was observed for the antero-posterior dimensions. The rate of margin re-excisions and completion mastectomies were lower in the OBS cohort. WLE patients with positive margins had a lower tumour-to-specimen ratio, whereas, the requirement for further surgery in the OBS cohort was associated with larger tumour dimensions.

Conclusion: Despite larger tumour dimensions, OBS is not inferior to WLE in providing clear surgical margins. Our analysis of the three-dimensional spatial relationship between cancer and surgical specimen, although not completely conclusive, can be helpful in the selection of the most appropriate surgical approach for every patient.
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http://dx.doi.org/10.1111/ans.16222DOI Listing
November 2020

Optimized Methods for the Production and Bioconjugation of Site-Specific, Alkyne-Modified Glucagon-like Peptide-1 (GLP-1) Analogs to Azide-Modified Delivery Platforms Using Copper-Catalyzed Alkyne-Azide Cycloaddition.

Bioconjug Chem 2020 07 25;31(7):1820-1834. Epub 2020 Jun 25.

School of Pharmacy, The University of Queensland, 20 Cornwall Street, Woolloongabba, Queensland 4102, Australia.

This study aimed to develop and optimize chemistries to produce alkyne-modified glucagon-like peptide-1(7-36)-amide (GLP-1(7-36)-NH) libraries, which could be rapidly and efficiently conjugated to other components and screened to identify compounds with the best drug delivery properties, as potential treatments for type 2 diabetes or obesity. For this purpose, the Lys26 (K26) side-chain, and the amino (N)- and carboxy (C)-termini of a dipeptidyl peptidase 4 (DPPIV)-resistant GLP-1 sequence (GLP-1(7-36;A8G)-NH), were modified with an alkyne (4-pentynoic acid or propiolic acid). These analogs were characterized with respect to human GLP-1 receptor (hGLP-1R) agonist activity, effects on cell viability and human serum stability, revealing that these modifications maintained low (N-terminal; EC 1.5 × 10 M) to subnanomolar (C-terminal and K26, ∼4 × 10 M) agonist activity toward hGLP-1, had no effect on cell viability, and for the N-terminal and K26 modifications, increased human serum proteolytic stability ( > 24 h). Copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction conditions were investigated using the C-terminal modified GLP-1 analog and an azide-modified model lipid peptide, with respect to the effects of altering the azide/alkyne ratio, cosolvents, temperature, reducing agents, Cu(I)-stabilizing ligand, copper source, and the concentrations of reagents/reactants, in order to identify general conditions that provide fast reactions and high yields. A 1:2 azide-alkyne (lipid:GLP-1 peptide) and 4:1 sodium ascorbate/copper sulfate molar ratio in 65% v/v DMSO-water at room temperature, in the absence of Cu(I)-stabilizing ligands (THPTA or l-histidine) and buffers (phosphate, pH 7), provided the best yields. This work reports a library of characterized GLP-1 analogs and chemistries for their attachment to other species, providing useful tools to improve GLP-1 delivery and pharmacology (e.g., through conjugation to other species that lower blood glucose, increase the duration of action, or enable delivery via a nonparenteral route).
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http://dx.doi.org/10.1021/acs.bioconjchem.0c00291DOI Listing
July 2020

Neutralisation of adeno-associated virus transduction by human vitreous humour.

Gene Ther 2021 May 15;28(5):242-255. Epub 2020 Jun 15.

LVF Ophthalmology Research Centre, Translational Research Institute, Woolloongabba, QLD, 4102, Australia.

Neutralising antibodies (NAbs), caused by past adeno-associated virus (AAV) infection, represent a critical challenge for AAV-mediated gene therapy, with even low NAb titres capable of inhibiting gene transfer, however in protein-rich environments such as the vitreous it is expected that other constituents could also interact with the transduction process. Inhibition of AAV2/2, AAV2/5, AAV2/6 and AAV2/8 transduction by human vitreous humour (VH) obtained from 80 post-mortem eye cups was investigated in this report, with clinically relevant vitreous dilutions as low as 1:2. Unexpectedly, the highest prevalence of inhibition of transduction was observed against AAV2/6, with 66% of tested samples displaying neutralisation at a 1:2 VH dilution. Only two samples showed inhibition of AAV2/8, indicating this serotype is an attractive vector for use in non-vitrectomised eyes of unscreened individuals. Levels of anti-AAV NAbs observed in the VH were much lower than previously observed in serum of a similar Australian population. Among ten tested eye cup pairs, we observed only small variation in anti-AAV NAbs levels between the left and right eye cups. Interaction with 1:2 diluted VH had an augmentation effect on AAV2/8 transduction (p = 0.004), a phenomenon which was not due to albumin or transferrin and which, if developed, might benefit the use of AAV2/8 in clinical settings.
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http://dx.doi.org/10.1038/s41434-020-0162-8DOI Listing
May 2021

Supercritical fluid assembly of albendazole liposomes targeting gastrin-releasing peptide receptor overexpressing tumors.

Nanomedicine (Lond) 2020 Jun 2. Epub 2020 Jun 2.

School of Pharmacy, The University of Queensland, Woolloongabba, QLD, 4102, Australia.

To develop albendazole (ABZ)-loaded bombesin(6-14) (BBN(6-14)) functionalized liposomes for targeting GRPR to enhance delivery to cancer cells. ABZ-loaded liposomes were formulated using supercritical CO technology; functionalized with a GRPR-targeted lipid-anchored BBN(6-14) peptide; and evaluated for effects on cell viability, particle size and targeted cell uptake. BBN(6-14)-coated ABZ liposomes decreased cell viability compared with nonfunctionalized ABZ liposomes. The level of GRPR expression positively correlated with intracellular uptake and decreased cell viability. The reduced cell viability, higher cell uptake and GRPR expression were observed in the order PC-3 > Caco-2 > HepG2 cells. BBN(6-14)-functionalized ABZ liposomes showed enhanced reduction in cell viability compared with nonfunctionalized ABZ liposomes.
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http://dx.doi.org/10.2217/nnm-2020-0048DOI Listing
June 2020

Severe meningoencephalitis secondary to calvarial invasion of forma in a dog.

Open Vet J 2020 04 19;10(1):31-38. Epub 2020 Feb 19.

Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611-0880, USA.

Background: The oomycete forma is an uncommon cause of severe dermal and subcutaneous infections in dogs with possible vascular invasion and other fatal sequelae. Infection within the central nervous system of affected dogs has not been previously reported.

Case Description: A 6-year-old spayed female mixed-breed dog was evaluated at a referral institution with a 2-month history of suspected fungal infection in the region of the right mandibular lymph node that was refractory to surgical resection and empiric medical therapy. Physical examination identified a 6-cm fluctuant subcutaneous mass caudoventral to the ramus of the right mandible and a second firm mass in the region of the right caudal maxilla. Lesional punch biopsies were submitted for fungal culture and polymerase chain reaction (PCR), which subsequently identified forma infection. Initial treatment consisted of anti-inflammatory doses of prednisone and hyperbaric oxygen therapy. Four weeks following initial evaluation, the patient was presented with progressive neurological signs consistent with a forebrain lesion. Magnetic resonance imaging revealed soft-tissue, contrast-enhancing lesions ventral to the calvarium adjacent to the site of original surgical resection and throughout the brain. Humane euthanasia was elected, and postmortem examination was consistent with the extension of local disease from the right masseter muscle into the right ventral calvarium. Postmortem DNA sequencing confirmed the identity of the organism as forma .

Conclusion: This is the first reported case of intracranial lagenidiosis in the dog. PCR distinguished this species from other species and from oomycetes of other genera, such as and . Regional extension of cutaneous lagenidiosis should therefore be considered in cases with concurrent or spontaneous neurologic disease.
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http://dx.doi.org/10.4314/ovj.v10i1.6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7193878PMC
April 2020

Development of an Enzyme-Mediated, Site-Specific Method to Conjugate Toll-Like Receptor 2 Agonists onto Protein Antigens: Toward a Broadly Protective, Four Component, Group A Streptococcal Self-Adjuvanting Lipoprotein-Fusion Combination Vaccine.

ACS Infect Dis 2020 07 1;6(7):1770-1782. Epub 2020 Jun 1.

School of Pharmacy, The University of Queensland, 20 Cornwall Street, Woolloongabba, Queensland 4102, Australia.

Subunit vaccines composed of protein antigens covalently attached to Toll-like receptor (TLR) agonists elicit superior immune responses compared to mixtures of antigens and TLR agonists. Among different conjugation approaches, enzyme-mediated ligation is one of the few that provides an opportunity for the generation of homogeneous, molecularly defined products in which protein antigens are maintained with native structures, which is most critical to elicit protective immune responses upon vaccination. Four highly conserved protein antigens from Group A (GAS) have the potential to be safe and efficacious vaccine candidates. After a TLR2 agonist fibroblast-stimulating lipopeptide-1 (FSL-1) was successfully attached onto each antigen using sortase A and techniques for their purification were developed, a combination vaccine containing interleukin 8 (IL-8) protease ( cell envelope proteinase [SpyCEP]), Group A Streptococcal C5a peptidase (SCPA), anchorless virulence factor arginine deiminase (ADI), and trigger factor (TF)-TLR2 conjugates was produced. This combination was assessed for immunity in mice and compared with mixtures of the four antigens with FSL-1 or alum. High titer antigen-specific IgG antibodies were detected from all vaccine groups, with antibodies elicited from FSL-1 conjugates around 10-fold higher compared to the FSL-1 mixture group. Furthermore, the FSL-1 conjugates afforded a more balanced T1/T2 immune response than the alum-adjuvanted group, suggesting that this combination vaccine represents a promising candidate for the prevention of GAS diseases. Thus, we established a conjugation platform that allows for the production of defined, site-specific antigen-adjuvant conjugates, which maintain the native three-dimensional structure of antigens and can be potentially applied to a variety of protein antigens.
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http://dx.doi.org/10.1021/acsinfecdis.0c00047DOI Listing
July 2020

A Self-Adjuvanting Vaccine Platform: Optimization of Site-Specific Sortase A Mediated Conjugation of Toll-Like Receptor 2 Ligands onto the Carboxyl or Amino terminus of Recombinant Protein Antigens.

Chempluschem 2020 01;85(1):227-236

Pharmacy Australia Centre of Excellence School of Pharmacy, The University of Queensland, 20 Cornwall St, Woolloongabba, QLD 4102, Australia.

Self-adjuvanting vaccines, consisting of recombinant protein antigens and covalently attached Toll-like receptor (TLR) agonists, have the ability to simultaneously and efficiently deliver antigen and TLR adjuvant to antigen presenting cells (APCs). Here, an enzyme-mediated ligation approach was used to overcome difficulties in producing homogeneous, molecularly defined self-adjuvanting vaccine products under native conditions. This process was optimized to allow the incorporation of the lipopeptide TLR2 agonist fibroblast-stimulating lipopeptide (FSL)-1 onto the N- or C-termini of recombinant protein antigens, employing the enzyme Staphylococcus aureus sortase A (SrtAsa) penta mutant. In addition, because SrtAsa-mediated ligations are reversible, a tryptophan zipper derived sequence was introduced into both reactants, which was demonstrated to improve ligation efficiency through the formation of a β-hairpin structure that hinders the reverse reaction. Finally, it was demonstrated that N- or C-terminal conjugation, and the incorporation of the β-hairpin structure, did not affect the TLR2-agonist activities of protein antigen TLR agonist conjugates. Overall, this SrtAsa-mediated ligation platform enabled production of antigen TLR2 agonist conjugates with enhanced ligation efficiency, with the conjugates demonstrating potent TLR2 signaling activation (EC <1nM).
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http://dx.doi.org/10.1002/cplu.201900687DOI Listing
January 2020

Semisynthetic, self-adjuvanting vaccine development: Efficient, site-specific sortase A-mediated conjugation of Toll-like receptor 2 ligand FSL-1 to recombinant protein antigens under native conditions and application to a model group A streptococcal vaccine.

J Control Release 2020 01 21;317:96-108. Epub 2019 Nov 21.

School of Pharmacy, The University of Queensland, Woolloongabba 4102, QLD, Australia; Australian Infectious Diseases Research Centre and School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD, Australia. Electronic address:

Protein antigens are, in general, weakly immunogenic, and therefore require co-delivery with adjuvants to stimulate potent immune responses. The fusion of (poly)peptide antigens to immunostimulatory adjuvants (e.g. Toll-like receptor (TLR) agonists) has been demonstrated to greatly improve vaccine potency compared to mixtures of antigen and adjuvant. Chemical approaches, to enable the rapid, site-specific and high-yielding linkage of TLR2 ligands to recombinant protein antigens, have been previously optimized. These approaches require the use of denaturing conditions to ensure high reaction yields, which limits their application, as maintenance of native protein folding is necessary to elicit antibodies against conformational epitopes. Here, this work aimed to optimize an alternative method, to ensure the efficient bioconjugation of TLR2 ligands onto folded protein antigens. An enzyme-mediated approach, using Staphylococcus aureus sortase A (or a penta mutant with enhanced efficiency), was optimized for reaction yield and time, as well as enzyme type and amount. This approach enabled the site-specific conjugation of the TLR2-agonist fibroblast-stimulating lipopeptide-1 (FSL-1) onto a model group A Streptococcus (GAS) recombinant polytope antigen under conditions that maintain protein folding, yielding a homogeneous, molecularly-defined product, with ligation yields as high as 90%. Following intramuscular (IM) administration of the ligation product to humanized plasminogen AlbPLG1 mice, high-titer, antigen-specific IgG antibodies were observed, which conferred protection against subcutaneous challenge with GAS strain 5448. In comparison, mixtures of the GAS antigen with aluminum hydroxide or FSL-1 failed to provide protection, with the FSL-1 mixture yielding ~1000-fold lower antigen-specific IgG antibody titers, and the mixture with alum yielding a Th2-biased response compared to the more balanced Th1/Th2 responses observed with the FSL-1 conjugate. Overall, a FSL-1 bioconjugation method for the efficient production of antigen-TLR2 agonist conjugates, which maintain protein folding, was produced, with broad utility for the development of self-adjuvanting vaccines against subunit protein antigens.
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http://dx.doi.org/10.1016/j.jconrel.2019.11.018DOI Listing
January 2020

Phenotypic Analysis of Human Lymph Nodes in Subjects With New-Onset Type 1 Diabetes and Healthy Individuals by Flow Cytometry.

Front Immunol 2019 31;10:2547. Epub 2019 Oct 31.

Department of Immunobiology, School of Immunology & Microbial Sciences (SIMS), King's College London, London, United Kingdom.

Ultrasound guided sampling of human lymph node (LN) combined with advanced flow cytometry allows phenotypic analysis of multiple immune cell subsets. These may provide insights into immune processes and responses to immunotherapies not apparent from analysis of the blood. Ultrasound guided inguinal LN samples were obtained by both fine needle aspiration (FNA) and core needle biopsy in 10 adults within 8 weeks of diagnosis of type 1 diabetes (T1D) and 12 age-matched healthy controls at two study centers. Peripheral blood mononuclear cells (PBMC) were obtained on the same occasion. Samples were transported same day to the central laboratory and analyzed by multicolour flow cytometry. LN sampling was well-tolerated and yielded sufficient cells for analysis in 95% of cases. We confirmed the segregation of CD69 cells into LN and the predominance of CD8 Temra cells in blood previously reported. In addition, we demonstrated clear enrichment of CD8 naïve, FOXP3 Treg, class-switched B cells, CD56 NK cells and plasmacytoid dendritic cells (DC) in LNs as well as CD4 T cells of the Th2 phenotype and those expressing Helios and Ki67. Conventional NK cells were virtually absent from LNs as were Th22 and Th1Th17 cells. Paired correlation analysis of blood and LN in the same individuals indicated that for many cell subsets, especially those associated with activation: such as CD25 and proliferating (Ki67) T cells, activated follicular helper T cells and class-switched B cells, levels in the LN compartment could not be predicted by analysis of blood. We also observed an increase in Th1-like Treg and less proliferating (Ki67) CD4 T cells in LN from T1D compared to control LNs, changes which were not reflected in the blood. LN sampling in humans is well-tolerated. We provide the first detailed "roadmap" comparing immune subsets in LN vs. blood emphasizing a role for differentiated effector T cells in the blood and T cell regulation, B cell activation and memory in the LN. For many subsets, frequencies in blood, did not correlate with LN, suggesting that LN sampling would be valuable for monitoring immuno-therapies where these subsets may be impacted.
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http://dx.doi.org/10.3389/fimmu.2019.02547DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6842967PMC
October 2020

Soil bacterial diffusible and volatile organic compounds inhibit Phytophthora capsici and promote plant growth.

Sci Total Environ 2019 Nov 5;692:267-280. Epub 2019 Jul 5.

Plant-Microbe Interactions Laboratory, School of Agriculture and Food Sciences, The University of Queensland, Brisbane, Queensland 4072, Australia.

Biotic interactions through diffusible and volatile organic compounds (VOCs) are frequent in nature. Soil bacteria are well-known producers of a wide range of volatile compounds (both organic and inorganic) with various biologically relevant activities. Since the last decade, they have been identified as natural biocontrol agents. Volatiles are airborne chemicals, which when released by bacteria, can trigger plant responses such as defence and growth promotion. In this study, we tested whether diffusible and volatile organic compounds (VOCs) produced by soil bacterial isolates exert anti-oomycete and plant growth-promoting effects. We also investigated the effects of inoculation with VOC-producing bacteria on the growth and development of Capsicum annuum and Arabidopsis thaliana seedlings. Our results demonstrate that organic VOCs emitted by bacterial antagonists negatively influence mycelial growth of the soil-borne phytopathogenic oomycete Phytophthora capsici by 35% in vitro. The bacteria showed plant growth promoting effects by stimulating biomass production, primary root growth and root hair development. Additionally, we provide evidence to suggest that these activities were deployed by the emission of either diffusible organic compounds or VOCs. Bacterial VOC profiles were obtained through solid phase microextraction (SPME) and analysis by gas chromatography coupled with mass spectrometry (GC-MS). This elucidated the main volatiles emitted by the isolates, which covered a wide range of aldehydes, alcohols, esters, carboxylic acids, and ketones. Collectively, twenty-five VOCs were identified to be produced by three bacteria; some being species-specific. Our data show that bacterial volatiles inhibits P. capsici in vitro and modulate both plant growth promotion and root system development. These results confirm the significance of soil bacteria and highlights that ways of harnessing them to improve plant growth, and as a biocontrol agent for soil-borne oomycetes through their volatile emissions deserve further investigation.
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http://dx.doi.org/10.1016/j.scitotenv.2019.07.061DOI Listing
November 2019

Diffusion kurtosis MRI as a predictive biomarker of response to neoadjuvant chemotherapy in high grade serous ovarian cancer.

Sci Rep 2019 07 24;9(1):10742. Epub 2019 Jul 24.

Department of Radiology, Box 218, University of Cambridge, Cambridge, CB2 0QQ, United Kingdom.

This study assessed the feasibility of using diffusion kurtosis imaging (DKI) as a measure of tissue heterogeneity and proliferation to predict the response of high grade serous ovarian cancer (HGSOC) to neoadjuvant chemotherapy (NACT). Seventeen patients with HGSOC were imaged at 3 T and had biopsy samples taken prior to any treatment. The patients were divided into two groups: responders and non-responders based on Response Evaluation Criteria In Solid Tumours (RECIST) criteria. The following imaging metrics were calculated: apparent diffusion coefficient (ADC), apparent diffusion (D) and apparent kurtosis (K). Tumour cellularity and proliferation were quantified using histology and Ki-67 immunohistochemistry. Mean K before therapy was higher in responders compared to non-responders: 0.69 ± 0.13 versus 0.51 ± 0.11 respectively, P = 0.02. Tumour cellularity correlated positively with K (rho = 0.50, P = 0.04) and negatively with both ADC (rho = -0.72, P = 0.001) and D (rho = -0.80, P < 0.001). Ki-67 expression correlated with K (rho = 0.53, P = 0.03) but not with ADC or D. In conclusion, K was found to be a potential predictive biomarker of NACT response in HGSOC, which suggests that DKI is a promising clinical tool for use oncology and radiology that should be evaluated further in future larger studies.
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http://dx.doi.org/10.1038/s41598-019-47195-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6656714PMC
July 2019

Climate vulnerability assessment for Pacific salmon and steelhead in the California Current Large Marine Ecosystem.

PLoS One 2019 24;14(7):e0217711. Epub 2019 Jul 24.

Department of Ecology and Evolutionary Biology, University of California, Santa Cruz, California, United States of America.

Major ecological realignments are already occurring in response to climate change. To be successful, conservation strategies now need to account for geographical patterns in traits sensitive to climate change, as well as climate threats to species-level diversity. As part of an effort to provide such information, we conducted a climate vulnerability assessment that included all anadromous Pacific salmon and steelhead (Oncorhynchus spp.) population units listed under the U.S. Endangered Species Act. Using an expert-based scoring system, we ranked 20 attributes for the 28 listed units and 5 additional units. Attributes captured biological sensitivity, or the strength of linkages between each listing unit and the present climate; climate exposure, or the magnitude of projected change in local environmental conditions; and adaptive capacity, or the ability to modify phenotypes to cope with new climatic conditions. Each listing unit was then assigned one of four vulnerability categories. Units ranked most vulnerable overall were Chinook (O. tshawytscha) in the California Central Valley, coho (O. kisutch) in California and southern Oregon, sockeye (O. nerka) in the Snake River Basin, and spring-run Chinook in the interior Columbia and Willamette River Basins. We identified units with similar vulnerability profiles using a hierarchical cluster analysis. Life history characteristics, especially freshwater and estuary residence times, interplayed with gradations in exposure from south to north and from coastal to interior regions to generate landscape-level patterns within each species. Nearly all listing units faced high exposures to projected increases in stream temperature, sea surface temperature, and ocean acidification, but other aspects of exposure peaked in particular regions. Anthropogenic factors, especially migration barriers, habitat degradation, and hatchery influence, have reduced the adaptive capacity of most steelhead and salmon populations. Enhancing adaptive capacity is essential to mitigate for the increasing threat of climate change. Collectively, these results provide a framework to support recovery planning that considers climate impacts on the majority of West Coast anadromous salmonids.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0217711PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6655584PMC
February 2020

Peptide-based targeted polymeric nanoparticles for siRNA delivery.

Nanotechnology 2019 Oct 11;30(41):415604. Epub 2019 Jul 11.

School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, St. Lucia, QLD 4072, Australia. Department of Pharmaceutical Sciences and Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, 140 The Fenway, Boston, MA 02115, United States of America.

The development of polymer-based nanoparticulate delivery systems for siRNA is important for the clinical success of gene therapy. However, there are some major drawbacks that need to be overcome. Short interfering RNA (siRNA) has been investigated as a potential therapeutic drug to silence disease-associated genes, but its usage is limited due to the lack of effective and safe nanocarriers. In this study, DOPE-PEI, a nanoparticle consisting of the fusogenic lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) conjugated with low-molecular-weight, 600 Da, branched polyethylenimine (PEI) was produced and optimized for siRNA delivery. This delivery system was modified with other components such as 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)2000] (DOPE-PEG2K), DOPE-PEG3.4K-bombesin and 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine/1,2-dioleoyl-3-trimethylammonium-propane (DOPE/DOTAP) and tested on PC-3 cells. The conjugation of DOPE to PEI polymer (DOPE-PEI) improved the efficiency of PEI to deliver siRNA into the cytosol and knockdown genes, but demonstrated high toxicity. The addition of DOPE-PEG2K reduced cellular toxicity by masking the surface positive charge of the DOPE-PEI/siRNA complex, with the incorporation of a gastrin-releasing peptide receptor (GRPR) targeting peptide and DOPE/DOTAP components improving the cellular uptake of siRNA into targeted cells and the siRNA knockdown efficiency.
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http://dx.doi.org/10.1088/1361-6528/ab313dDOI Listing
October 2019

Comment on "Designing river flows to improve food security futures in the Lower Mekong Basin".

Science 2019 06;364(6444)

Aquae Sulis Research Ltd., Midway House, Turleigh, Wiltshire BA15 2LR, UK.

Sabo (Research Articles, 8 December 2017, p. 1270) used statistical relationships between flow and catch in a major Lower Mekong Basin fishery to propose a flow regime that they claim would increase catch, if implemented by proposed dams. However, their catch data were not adjusted for known variation in monitoring effort, invalidating their analysis.
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http://dx.doi.org/10.1126/science.aav8755DOI Listing
June 2019

Advances in Targeted Gene Delivery.

Curr Drug Deliv 2019 ;16(7):588-608

School of Pharmacy, The University of Queensland, Woolloongabba, 4102, Australia.

Gene therapy has the potential to treat both acquired and inherited genetic diseases. Generally, two types of gene delivery vectors are used - viral vectors and non-viral vectors. Non-viral gene delivery systems have attracted significant interest (e.g. 115 gene therapies approved for clinical trials in 2018; clinicaltrials.gov) due to their lower toxicity, lack of immunogenicity and ease of production compared to viral vectors. To achieve the goal of maximal therapeutic efficacy with minimal adverse effects, the cell-specific targeting of non-viral gene delivery systems has attracted research interest. Targeting through cell surface receptors; the enhanced permeability and retention effect, or pH differences are potential means to target genes to specific organs, tissues, or cells. As for targeting moieties, receptorspecific ligand peptides, antibodies, aptamers and affibodies have been incorporated into synthetic nonviral gene delivery vectors to fulfill the requirement of active targeting. This review provides an overview of different potential targets and targeting moieties to target specific gene delivery systems.
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http://dx.doi.org/10.2174/1567201816666190529072914DOI Listing
February 2020

Gastrin-releasing peptide receptor-targeted hybrid peptide/phospholipid pDNA/siRNA delivery systems.

Nanomedicine (Lond) 2019 05 3;14(9):1153-1171. Epub 2019 May 3.

School of Pharmacy, The University of Queensland, Woolloongabba 4102, Australia.

To develop a peptide/phospholipid hybrid system for gastrin-releasing peptide receptor (GRPR)-targeted delivery of pDNA or siRNA. A multifunctional GRPR-targeted peptide R-K(GALA)-BBN(6-14) was combined with a phospholipid oligonucleotide delivery system (1:1 1,2-dioleoyl--glycero-3-phosphoethanolamine and 1,2-dioleoyl-3-trimethylammonium-propane) and evaluated for pDNA and siRNA delivery in terms of complex size, toxicity, receptor-targeted delivery and gene expression or knockdown efficiency. By combining peptide and phospholipid delivery systems, synergistic improvements in gene expression and knockdown were observed when compared with either system alone. The optimized formulation demonstrated high levels of EGFP expression and EGFP knockdown, GRPR-targeted delivery, enhanced endosomal release and minimal toxicity. The peptide/phospholipid hybrid system provides efficient GRPR-targeted DNA/siRNA delivery.
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http://dx.doi.org/10.2217/nnm-2018-0380DOI Listing
May 2019

Glucagon-Like Peptide-1 Receptor Agonists and Strategies To Improve Their Efficiency.

Mol Pharm 2019 06 13;16(6):2278-2295. Epub 2019 May 13.

School of Pharmacy , The University of Queensland , Woolloongabba , 4102 , Australia.

Type 2 diabetes mellitus (T2DM) is increasing in global prevalence and is associated with serious health problems (e.g., cardiovascular disease). Various treatment options are available for T2DM, including the incretin hormone glucagon-like peptide-1 (GLP-1). GLP-1 is a therapeutic peptide secreted from the intestines following food intake, which stimulates the secretion of insulin from the pancreas. The native GLP-1 has a very short plasma half-life, owning to renal clearance and degradation by the enzyme dipeptidyl peptidase-4. To overcome this issue, various GLP-1 agonists with increased resistance to proteolytic degradation and reduced renal clearance have been developed, with several currently marketed. Strategies, such as controlled release delivery systems, methods to reduce renal clearance (e.g., PEGylation and conjugation to antibodies), and methods to improve proteolytic stability (e.g., stapling, cyclization, and glycosylation) provide means to further improve the ability of GLP-1 analogs. These will be discussed in this literature review.
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http://dx.doi.org/10.1021/acs.molpharmaceut.9b00308DOI Listing
June 2019

An Experimental Group A Vaccine That Reduces Pharyngitis and Tonsillitis in a Nonhuman Primate Model.

mBio 2019 04 30;10(2). Epub 2019 Apr 30.

Australian Infectious Diseases Research Centre, The University of Queensland, St Lucia, QLD, Australia

Group A (GAS) infections account for an estimated 500,000 deaths every year. This bacterial pathogen is responsible for a variety of mild and life-threatening infections and the triggering of chronic autoimmune sequelae. Pharyngitis caused by group A (GAS), but not asymptomatic GAS carriage, is a prerequisite for acute rheumatic fever (ARF). Repeated bouts of ARF may trigger rheumatic heart disease (RHD), a major cause of heart failure and stroke accounting for 275,000 deaths annually. A vaccine that prevents pharyngitis would markedly reduce morbidity and mortality from ARF and RHD. Nonhuman primates (NHPs) have been utilized to model GAS diseases, and experimentally infected rhesus macaques develop pharyngitis. Here we use an NHP model of GAS pharyngitis to evaluate the efficacy of an experimental vaccine, Combo5 (arginine deiminase [ADI], C5a peptidase [SCPA], streptolysin O [SLO], interleukin-8 [IL-8] protease [SpyCEP], and trigger factor [TF]), specifically designed to exclude GAS components potentially linked to autoimmune complications. Antibody responses against all Combo5 antigens were detected in NHP serum, and immunized NHPs showed a reduction in pharyngitis and tonsillitis compared to controls. Our work establishes the NHP model as a gold standard for the assessment of GAS vaccines. GAS-related diseases disproportionally affect disadvantaged populations (e.g., indigenous populations), and development of a vaccine has been neglected. A recent strong advocacy campaign driven by the World Health Organization and the International Vaccine Institute has highlighted the urgent need for a GAS vaccine. One significant obstacle in GAS vaccine development is the lack of a widely used animal model to assess vaccine efficacy. Researchers in the field use a wide range of murine models of infection and assays, sometimes yielding conflicting results. Here we present the nonhuman primate pharyngeal infection model as a tool to assess vaccine-induced protection against colonization and clinical symptoms of pharyngitis and tonsillitis. We have tested the efficacy of an experimental vaccine candidate with promising results. We believe that the utilization of this valuable tool by the GAS vaccine research community could significantly accelerate the realization of a safe and effective GAS vaccine for humans.
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http://dx.doi.org/10.1128/mBio.00693-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6495378PMC
April 2019

Sodium MRI with 3D-cones as a measure of tumour cellularity in high grade serous ovarian cancer.

Eur J Radiol Open 2019 19;6:156-162. Epub 2019 Apr 19.

Department of Radiology, University of Cambridge, Cambridge, CB2 0QQ, United Kingdom.

The aim of this study was to assess the feasibility of rapid sodium MRI (Na-MRI) for the imaging of peritoneal cancer deposits in high grade serous ovarian cancer (HGSOC) and to evaluate the relationship of Na-MRI with tumour cellularity. Na-MRI was performed at 3 T on twelve HGSOC patients using a 3D-cones acquisition technique. Tumour biopsies specimens were collected after imaging and cellularity was measured from histology. Total Na-MRI scan time for each patient was approximately 11 min. At an isotropic resolution of 5.6 mm, signal-to-noise ratios (SNRs) of 82.2 ± 15.3 and 15.1 ± 7.1 (mean ± standard deviation) were achieved for imaging of tumour tissue sodium concentration (TSC) and intracellular weighted sodium concentration (IWS) respectively. Tumour TSC and IWS concentrations were: 56.8 ± 19.1 mM and 30.8 ± 9.2 mM respectively and skeletal muscle TSC and IWS concentrations were 33.2 ± 16.3 mM and 20.5 ± 9.9 mM respectively. There were significant sodium concentration differences between cancer and skeletal muscle, Wilcoxon signed-rank test,  <  0.001 for TSC and  =  0.01 for IWS imaging. Tumour cellularity displayed a strong negative correlation with TSC, Spearman's rho = -0.92,  <  0.001, but did not correlate with IWS. This study demonstrates that Na-MRI using 3D-cones can rapidly assess sodium concentration in peritoneal deposits of HGSOC and that TSC may serve as a biomarker of tumour cellularity.
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http://dx.doi.org/10.1016/j.ejro.2019.04.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6477161PMC
April 2019

Dispersibility of phospholipids and their optimization for the efficient production of liposomes using supercritical fluid technology.

Int J Pharm 2019 May 30;563:174-183. Epub 2019 Mar 30.

School of Pharmacy, The University of Queensland, Woolloongabba, Brisbane, QLD 4102, Australia. Electronic address:

Liposomes are promising delivery vehicles and offer the added drawcard of being able to be made functional to target tissues such as cardiac muscle and cancerous cells. Current methods to manufacture liposomes need to be improved and supercritical fluid (SCF) technologies may offer a solution. Herein, the dispersibility of six different phospholipids (PLs) was determined in supercritical carbon dioxide (scCO). 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) showed the highest post-processing dispersibility, while 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) showed no dispersibility in scCO at the assessed experimental conditions. The zetasizer results showed that the SCF conditions at 37 °C, 250 bar and 200 RPM for 60 min provided nanoparticles with the narrowest polydispersity index (PDI) and a spherical shape as shown by cryo-transmission electron microscopy (Cryo-TEM). The mean diameter of liposomes using the SCF method for DSPC-PEGylated and DOPC-PEGylated liposomes was 98.3 ± 3.3 nm and 124.5 ± 4.1 nm, while using the thin film method it was 153.6 ± 4.5 nm and 131.3 ± 3.4 nm, respectively. A size-based stability evaluation of the scCO-prepared liposomes stored at different temperatures (25 °C, 4 °C and -20 °C) was compared to that of the thin film method over a period of 3 months. The current study provides a possible green alternative SCF method to preparing liposomes that is less laborious, time saving, and a low energy process.
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http://dx.doi.org/10.1016/j.ijpharm.2019.03.053DOI Listing
May 2019

Preparation of albendazole-loaded liposomes by supercritical carbon dioxide processing.

Artif Cells Nanomed Biotechnol 2018 28;46(sup3):S1186-S1192. Epub 2019 Jan 28.

a School of Pharmacy , The University of Queensland , Woolloongabba , Australia.

Supercritical fluid (SCF) technology offers a potential green alternative to organic solvent-based methods for drug formulation. Albendazole (ABZ) has promising anticancer activity when formulated to increase its cellular uptake. Herein, a static volume method was used to determine the solubility of ABZ in supercritical carbon dioxide (scCO) for the future development of such ABZ formulations. The solubility of ABZ in scCO (250 bar, 37 °C) was approximately 12 mg/100 mL. The extent of dissolution was measured at various time points to determine when saturation solubility occurred, which was demonstrated from 9 h. In order to determine if scCO processing induced ABZ polymorphism, DSC/TGA, FTIR and XRD were used, which demonstrated no change in its solid state. Following this, ABZ loaded liposomes were manufactured using SCF technology. The liposomes diameter was 167.2 ± 5.3 nm as determined by Zetasizer, and confirmed by cryo-transmission electron microscopy. In conclusion, scCO was used successfully to solubilize ABZ, and to manufacture liposomes of nano-sized range. This study provides insight into use of green technology for future ABZ liposomal formulation without the need for organic solvents.
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http://dx.doi.org/10.1080/21691401.2018.1536059DOI Listing
June 2019

A reappraisal of the California Roach/Hitch (Cypriniformes, Cyprinidae, Hesperoleucus/Lavinia) species complex.

Zootaxa 2019 Jan 6;4543(2):221-240. Epub 2019 Jan 6.

Center for Watershed Sciences, University of California, Davis, One Shields Avenue, Davis, CA 95616 Department of Animal Sciences, University of California, Davis, One Shields Avenue, Davis, CA 95616 Department of Biological Sciences, University of the Pacific, 3601 Pacific Ave, Stockton, CA 95211.

The California Roach (Hesperoleucus symmetricus) and Hitch (Lavinia exilicauda) form a species complex largely endemic to California (CA), USA. Using previous studies of this complex along with a recent comprehensive genomic analysis, we developed a highly supported taxonomic hierarchy of two genera, five species, four subspecies and multiple distinct population segments within two presently recognized species. The genera Lavinia and Hesperoleucus are supported as representing distinct lineages, despite occasional hybridization between them. While hybridization is one pathway to some speciation in this complex, hierarchical levels correlate nicely between genomic results and earlier morphological work. Hesperoleucus symmetricus is newly divided into four species (H. parvipinnis-Gualala Roach, H. mitrulus-Northern Roach, H. venustus-Coastal Roach, and H. symmetricus-California Roach) and two subspecies (H. s. serpentinus-Red Hills Roach, H. s. symmetricus-California Roach). Within H. venustus, two subspecies are identified (H. v. navarroensis-Northern Coastal Roach, and H. v. subditus-Southern Coastal Roach), which are supported by previous morphological studies but resolve discrepancies between those studies. Finally, six distinct population segments are identified within different species/subspecies: Kaweah, Russian River, Navarro River, Monterey, and Tomales Bay. Clear Lake Roach are introgressed between California and Coastal Roach, making them distinct but difficult to formally name. Results should greatly improve management and conservation of each taxonomic entity and help resolve past ambiguities. Additional studies are needed to improve range-wide boundaries and to investigate population structure within all species and subspecies identified in both Lavinia and Hesperoleucus lineages.
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http://dx.doi.org/10.11646/zootaxa.4543.2.3DOI Listing
January 2019

Effect of common storage temperatures and container types on urine protein : creatinine ratios in urine samples of proteinuric dogs.

J Vet Intern Med 2018 Sep 17;32(5):1652-1658. Epub 2018 Sep 17.

Department of Small Animal Clinical Sciences, University of Florida, College of Veterinary Medicine, Gainesville, Florida.

Background: Preanalytic protein adsorption to polymer and glass container surfaces may decrease urine protein concentration measurements and urine protein: creatinine ratios (UPC).

Hypothesis/objectives: Urine stored in PC or glass containers will have lower UPC than urine stored in HP containers. The specific objective was to determine whether clinically relevant differences in UPC would be detected after storage in glass, PC, or HP containers using common storage times and temperatures.

Animals: Twelve client-owned dogs with proteinuria.

Methods: Prospective, nonmasked study, divided into 2 phases. The first phase was a pilot study involving multiple (n = 5) measurements at each storage condition using 24-hours urine samples from 2 dogs with persistent renal proteinuria of different magnitude. The second phase used urine samples from 10 dogs with proteinuria of variable magnitude. Sample aliquots were stored in HP, PC, and glass containers at 24°C for 4 hours, 4°C for 12 hours, and -20°C for 72 hours. The UPC of each was measured after storage and compared with baseline.

Results: Statistically significant but clinically irrelevant differences were found in phase 1. In phase 2, storage conditions did not affect urinary protein or creatinine concentrations or UPC.

Conclusions And Clinical Importance: Collection and storage of canine urine samples in clean HP, PC, or glass containers at 24°C for 4 hours, 4°C for 12 hours, or -20°C for 72 hours is unlikely to result in clinically relevant decreases in measured UPC values.
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http://dx.doi.org/10.1111/jvim.15232DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6189361PMC
September 2018
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