Parkhouse RM et al. (1985) Clin Exp Immunol "Human macrophages and T-lymphocyte subsets infiltrating nodules of Onchocerca volvulus."

Janossy G, Parkhouse RM, Bofill M, Gomez-Priego A

[Clin Exp Immunol, 1985]

The nature of the lymphoid infiltrate in nodules of Onchocerca volvulus was assessed using monoclonal antibodies to lymphoid cell surface markers. Although B cells were generally absent, T cells were present, but in variable amounts. The ratio of T4+ (helper phenotype) to T8+ (suppressor-cytotoxic phenotype) was usually in the normal peripheral blood range of about 3, although ratios ranging from 1 to 10 were seen in selected areas of the onchocercoma. The possibility of immunosuppression through dominance of T4+, Leu-8+ cells (suppressor-inducer phenotype) within the T4+ population was also excluded. The T cells did not tend to concentrate in close proximity to the parasite, and there was no general bias in favour of the T suppressor cell phenotype (T8) within the infiltrate. Macrophages and dendritic cells were consistently observed and consisted of three defined cell types in approximately equal proportions: normal, unactivated macrophages (HLA-DR-, acid phosphatase positive), activated macrophages (HLA-DR+, acid phosphatase positive) and cells of dendritic morphology (HLA-DR+, acid phosphatase negative). These results are discussed in relation to immune suppression in filariasis.

Ma C et al. (2015) Molecules "Potent Activities of Roemerine against Candida albicans and the Underlying Mechanisms."

Yan L, Xu G, Wang C, He G, Du F, Jiang Y, Ma C, Rao G, He J

[Molecules, 2015]

Roemerine (RM) is an aporphine alkaloid isolated from the fresh rattan stem of Fibraurea recisa, and it has been demonstrated to have certain antifungal activity. This study aimed to investigate the antifungal activity of RM and the underlying mechanisms in Candida albicans (C. albicans). The in vitro antifungal activity of RM was evaluated by a series of experiments, including the XTT reduction assay, confocal laser scanning microscopy assay, scanning electron microscope assay. Results showed that 1 g/mL RM inhibited biofilm formation significantly (p < 0.01) both in Spider medium and Lee's medium. In addition, RM could inhibit yeast-to-hyphae transition of C. albicans in a dose-dependent manner. The biofilm-specific and hypha-specific genes such as YWP1, SAP5, SAP6, HWP1, ECE1 were up-regulated and EFG1 was down-regulated after 8 g/mL RM treatment. Furthermore, the toxicity of RM was investigated using C. elegans worms, three cancer cells and one normal cell. The date showed that RM had no significant toxicity. In conclusion, RM could inhibited the formation of C. albicans biofilm in vitro, but it had no fungicidal effect on planktonic C. albicans cells, and the anti-biofilm mechanism may be related to the cAMP pathway.

Cabrera Z et al. (1987) Parasite Immunol "Isolation of an antigenic fraction for diagnosis of onchocerciasis."

Cabrera Z, Parkhouse RM

[Parasite Immunol, 1987]

A surface enriched fraction was prepared from adults of Onchocerca volvulus by brief extraction of entire worms with detergent. This was then gel filtered to yield a low molecular weight fraction which functioned specifically in ELISA analysis. An identical result was also obtained when the related cattle parasite, O. gibsoni, was similarly fractionated and tested. The low molecular weight fraction contained at least four antigenic components when examined by coprecipitation and immunoblotting studies. One ml of packed worms yielded sufficient low molecular weight antigen to examine about 2,000 human sera by the ELISA procedure, and the test was sensitive at human serum dilutions down to 1/400. A preliminary study with individual sera from Onchocerciasis endemic and non-endemic areas of Southern Mexico yielded 0/24 false positives, 3/24 false negatives and a significant ELISA value in 21/24 sera from proven cases of Onchocerciasis.

Gouveia GJ et al. (2021) Anal Chem "Long-Term Metabolomics Reference Material."

Andersen EC, Morse AM, Garcia BM, Edison AS, Shaver AO, Gouveia GJ, McIntyre LM

[Anal Chem, 2021]

The use of quality control samples in metabolomics ensures data quality, reproducibility, and comparability between studies, analytical platforms, and laboratories. Long-term, stable, and sustainable reference materials (RMs) are a critical component of the quality assurance/quality control (QA/QC) system; however, the limited selection of currently available matrix-matched RMs reduces their applicability for widespread use. To produce an RM in any context, for any matrix that is robust to changes over the course of time, we developed iterative batch averaging method (IBAT). To illustrate this method, we generated 11 independently grown <i>Escherichia coli</i> batches and made an RM over the course of 10 IBAT iterations. We measured the variance of these materials by nuclear magnetic resonance (NMR) and showed that IBAT produces a stable and sustainable RM over time. This <i>E. coli</i> RM was then used as a food source to produce a <i>Caenorhabditis elegans</i> RM for a metabolomics experiment. The metabolite extraction of this material, alongside 41 independently grown individual <i>C. elegans</i> samples of the same genotype, allowed us to estimate the proportion of sample variation in preanalytical steps. From the NMR data, we found that 40% of the metabolite variance is due to the metabolite extraction process and analysis and 60% is due to sample-to-sample variance. The availability of RMs in untargeted metabolomics is one of the predominant needs of the metabolomics community that reach beyond quality control practices. IBAT addresses this need by facilitating the production of biologically relevant RMs and increasing their widespread use.

Lawal TA et al. (2020) Orphanet J Rare Dis "Preclinical model systems of ryanodine receptor 1-related myopathies and malignant hyperthermia: a comprehensive scoping review of works published 1990-2019."

Dowling JJ, Terry NL, Todd JJ, Wires ES, Lawal TA

[Orphanet J Rare Dis, 2020]

BACKGROUND: Pathogenic variations in the gene encoding the skeletal muscle ryanodine receptor (RyR1) are associated with malignant hyperthermia (MH) susceptibility, a life-threatening hypermetabolic condition and RYR1-related myopathies (RYR1-RM), a spectrum of rare neuromuscular disorders. In RYR1-RM, intracellular calcium dysregulation, post-translational modifications, and decreased protein expression lead to a heterogenous clinical presentation including proximal muscle weakness, contractures, scoliosis, respiratory insufficiency, and ophthalmoplegia. Preclinical model systems of RYR1-RM and MH have been developed to better understand underlying pathomechanisms and test potential therapeutics. METHODS: We conducted a comprehensive scoping review of scientific literature pertaining to RYR1-RM and MH preclinical model systems in accordance with the PRISMA Scoping Reviews Checklist and the framework proposed by Arksey and O'Malley. Two major electronic databases (PubMed and EMBASE) were searched without language restriction for articles and abstracts published between January 1, 1990 and July 3, 2019. RESULTS: Our search yielded 5049 publications from which 262 were included in this review. A majority of variants tested in RYR1 preclinical models were localized to established MH/central core disease (MH/CCD) hot spots. A total of 250 unique RYR1 variations were reported in human/rodent/porcine models with 95% being missense substitutions. The most frequently reported RYR1 variant was R614C/R615C (human/porcine total n=39), followed by Y523S/Y524S (rabbit/mouse total n=30), I4898T/I4897T/I4895T (human/rabbit/mouse total n=20), and R163C/R165C (human/mouse total n=18). The dyspedic mouse was utilized by 47% of publications in the rodent category and its RyR1-null (1B5) myotubes were transfected in 23% of publications in the cellular model category. In studies of transfected HEK-293 cells, 57% of RYR1 variations affected the RyR1 channel and activation core domain. A total of 15 RYR1 mutant mouse strains were identified of which ten were heterozygous, three were compound heterozygous, and a further two were knockout. Porcine, avian, zebrafish, C. elegans, canine, equine, and drosophila model systems were also reported. CONCLUSIONS: Over the past 30years, there were 262 publications on MH and RYR1-RM preclinical model systems featuring more than 200 unique RYR1 variations tested in a broad range of species. Findings from these studies have set the foundation for therapeutic development for MH and RYR1-RM.

Harnett W et al. (1989) Mol Biochem Parasitol "An oligonucleotide probe specific for Onchocerca volvulus."

Harnett W, Chambers AE, Renz A, Parkhouse RM

[Mol Biochem Parasitol, 1989]

A genomic DNA library of a Liberian strain of Onchocerca volvulus was prepared in the vector bacteriophage lambda gt10. The library was differentially screened by hybridisation with radiolabelled total DNA from the homologous parasite, two heterologous Onchocerca parasites (Onchocerca gibsoni and Onchocerca gutturosa) and human liver cells. A clone (C1A1) was isolated whose binding to O. volvulus DNA was at least 50 times stronger than to the other parasite DNA samples. No binding was observed with human DNA. The insert of C1A1 was subcloned into the filamentous phage vector M13 mp18 and sequenced. Two oligonucleotides, each corresponding to a unique region of 60 nucleotides (out of a total of 154) were synthesised and examined for hybridisation with three different geographical isolates of O. volvulus (including forest and savannah strains) and six other Onchocerca spp. One of the oligonucleotides (C1A1-2) was found to hybridise to the three O. volvulus isolates with an intensity in the region of 300 times greater than to any other Onchocerca spp. Since the other species include the two which may be most closely related to O. volvulus, i.e., O. gibsoni and Onchocerca ochengi, it is concluded that C1A1-2 is likely to represent a truly species-specific probe.

Garate T et al. (1990) Trop Med Parasitol "Cloning of specific diagnostic antigens of Onchocerca volvulus."

Parkhouse RM, Harnett W, Conraths FJ, Buttner DW, Garate T

[Trop Med Parasitol, 1990]

Specific, serological diagnosis is one of the main goals in onchocerciasis research. To date this objective has been hampered by (a) scarcity of parasite material, and (b) antigenic cross-reaction between Onchocerca volvulus and other nematode species. In order to obtain specific antigens, and in amounts suitable for study, molecular biological techniques have been adopted. A lambda gt11 cDNA expression library prepared from O. volvulus adult female worms was screened using infected human sera from onchocerciasis patients and rabbit hyperimmune sera raised against Onchocerca and genus-specific Onchocerca antigen extracts. Five clones were selected and their inserts expressed as beta-galactosidase fusion proteins. The fusion proteins were examined using individual sera from patients with O. volvulus or Wuchereria bancrofti infections. Three of the fusion proteins were recognised by more than 80% of O. volvulus sera and exhibited weak reactivity with a few W. bancrofti sera. One of these three clones was recognised to a significantly greater degree by sera from sowda than from generalised onchocerciasis patients.

Volpatti JR et al. (2020) Elife "Identification of drug modifiers for RYR1 related myopathy using a multi-species discovery pipeline."

Groom L, Endo Y, Volpatti JR, Zuercher WJ, Brennan S, Roy P, Dirksen RT, Dowling JJ, Knox J, Noche R

[Elife, 2020]

Ryanodine receptor type I-related myopathies (RYR1-RMs) are a common group of childhood muscle diseases associated with severe disabilities and early mortality for which there are no available treatments. The goal of this study is to identify new therapeutic targets for RYR1-RMs. To accomplish this, we developed a discovery pipeline using nematode, zebrafish, and mammalian cell models. We first performed large-scale drug screens in <i>C. elegans</i> which uncovered 74 hits. Targeted testing in zebrafish yielded positive results for two p38 inhibitors. Using mouse myotubes, we found that either pharmacological inhibition or siRNA silencing of p38 impaired caffeine-induced Ca²⁺ release from wild type cells while promoting intracellular Ca²⁺ release in <i>Ryr1</i> knockout cells. Lastly, we demonstrated that p38 inhibition blunts the aberrant temperature-dependent increase in resting Ca²⁺ in myotubes from an RYR1-RM mouse model. This unique platform for RYR1-RM therapy development is potentially applicable to a broad range of neuromuscular disorders.

Cabrera Z et al. (1989) Trop Med Parasitol "Specific detection of human antibodies to Onchocerca volvulus."

Cabrera Z, Yarzabal L, Forsyth K, Pabon R, Gomez Priego A, Parkhouse RM

[Trop Med Parasitol, 1989]

Specific diagnosis of antibodies to Onchocerca was achieved through (1) the construction of direct and indirect ELISA systems, and (2) restricting ELISA assays to the IgG4 class. The direct ELISA was based on the isolation of a surface derived, low molecular weight surface antigen preparation containing two main antigens (M. wt. 16.2 and 12.8 kDA) as defined by Western blot analysis. The direct ELISA system detected antibodies in children of six years old, and may therefore be applicable to detecting reinvasion in OCP areas of Onchocerca volvulus control. The indirect ELISA system was a competitive binding ELISA-based assay using a monoclonal antibody recognising two Onchocerca components (M. wts. 15.6 and 25.9) on a Western blot. The direct and indirect ELISA systems were similarly specific and sensitive when evaluated in a preliminary survey. The direct ELISA system yielded a specificity and sensitivity of: 100% and 100% respectively, using Mexican endemic and Mexican intestinal nematode infection sera as positive and negative controls respectively: 91% and 96% respectively, using Venezuelan endemic and Venezuelan Mansonella ozzardi infection sera as positive and negative controls, respectively: 87% and 93% respectively, using African endemic and Papuan (New Guinea) Wuchereria bancrofti infection sera as positive and negative controls respectively: 93% and 93% respectively, using African endemic and Indian W. bancrofti infection sera as positive and negative controls respectively. Similar specificity and sensitivity levels were obtained when the same comparisons were made using the indirect (inhibition) ELISA assay. These values may be contrasted with the currently used PBS extract of O. volvulus which yielded specificities of less than 10% in all the above comparisons.(ABSTRACT TRUNCATED AT 250 WORDS)

Yu Z et al. (2011) J Environ Sci (China) "Behavior toxicity to Caenorhabditis elegans transferred to the progeny after exposure to sulfamethoxazole at environmentally relevant concentrations."

Yu Z, Jiang L, Yin D

[J Environ Sci (China), 2011]

Sulfamethoxazole (SMX) is one of the most common detected antibiotics in the environment. In order to study whether SMX can affect behavior and growth and whether these effects could be transferred to the progeny, Caenorhabditis elegans was exposed at environmentally relevant concentrations for 24, 48, 72 and 96 hr, respectively. After exposure, the exposed parent generation (P0) was measured for behavior and growth indicators, which were presented as percentage of controls (POC). Then their corresponding unexposed progeny (F1) was separated and measured for the same indicators. The lowest POC for P0 after 96 hr-exposure at 100 mg/L were 37.8%, 12.7%, 45.8% and 70.1% for body bending frequency (BBF), reversal movement (RM), Omega turns (OT) and body length (BL), respectively. And F1 suffered defects with the lowest POC as 55.8%, 24.1%, 48.5% and 60.7% for BBF, RM, OT and BL, respectively. Defects in both P0 and F1 showed a time- and concentration-dependent fashion and behavior indicators showed better sensitivity than growth indicator. The observed effects on F1 demonstrated the transferable properties of SMX. Defects of SMX at environmental concentrations suggested that it is necessary to perform further systematical studies on its ecological risk in actual conditions.