Prevalence of the ABCB1-1Δ gene mutation in a sample of New Zealand Huntaway dogs.

AIMS: To determine the prevalence of the ATP Binding Cassette Subfamily B Member 1-1Δ mutation (ABCB1-1Δ; previously Multidrug Resistance 1 (MDR1) mutation) in a cohort of New Zealand Huntaway dogs. MATERIALS AND METHODS: Samples were opportunistically collected from Huntaway dogs (n = 189) from throughout New Zealand. Buccal swabs were collected from 42 Huntaways from the Wairarapa region and 147 blood samples from Huntaways from the Gisborne, Waikato, Manawatu/Whanganui, Hawkes Bay, Canterbury and Otago regions. DNA was extracted from all samples and tested for the presence of the ABCB1-1Δ allele. RESULTS: Of 189 Huntaway dogs that were tested, two were found to be heterozygous carriers of the ABCB1-1Δ allele and the remaining 187/189 dogs were homozygous for the wild type allele. No dogs homozygous for the mutation were identified. CONCLUSIONS AND CLINICAL RELEVANCE: The results of this study show that the ABCB1-1Δ allele is present in Huntaway dogs. The low prevalence in this convenience sample suggests that the prevalence of this allele in the Huntaway population is likely to be low. We recommend that veterinary clinicians discuss the potential for this mutation in Huntaways with dog owners including the clinical implications for dogs that are homozygous for the mutated allele and the potential for testing for the mutation, as they would do for other known mutations.

Predictors of social participation: evidence from repeated cross-sectional population surveys in England.

BACKGROUND: Social participation is linked to better health and well-being. However, there is limited research on the individual and area-level predictors of participation. This study aims to determine the characteristics associated with participation, particularly the impact of community asset availability. METHODS: We used data from 34 582 adult respondents to the nationally representative Community Life Survey from 2013 to 2018. We measured social participation by reported participation in 15 types of groups. We used probit and negative binomial regression models and included a wide range of individual, household and area characteristics, and availability of 14 types of community assets. RESULTS: The following characteristics were associated with higher levels of participation: being female (+3.0 percentage points (p.p.) (95% CI 1.8 to 4.1p.p.), Black, Asian or Minority Ethnicity (+3.7p.p. (1.9 to 5.5p.p.)), homeownership (+4.1 p.p. (2.7 to 5.6p.p.)) and living in a rural area (+2.1p.p. (0.5 to 3.6p.p)). Respondents from the most deprived areas were less likely to participate than those in average deprivation areas (-3.9p.p. (-5.9 to -1.99p.p.)). Higher availability of community assets was associated with increased participation in groups. The effect of availability on participation varied by type of asset. CONCLUSION: Improving community assets infrastructure in high deprivation and urban areas would encourage more social participation in these areas.

Cyclin-dependent kinase 7 is a therapeutic target in high-grade glioma.

High-grade glioma (HGG) is an incurable brain cancer. The transcriptomes of cells within HGG tumors are highly heterogeneous. This renders the tumors unresponsive or able to adapt to therapeutics targeted at single pathways, thereby causing treatment failure. To overcome this, we focused on cyclin-dependent kinase 7 (CDK7), a ubiquitously expressed molecule involved in two major drivers of HGG pathogenesis: cell cycle progression and RNA polymerase-II-based transcription. We tested the activity of THZ1, an irreversible CDK7 inhibitor, on patient-derived primary HGG cell lines and ex vivo HGG patient tissue slices, using proliferation assays, microarray analysis, high-resolution respirometry, cell cycle analysis and in vivo tumor orthografts. The cellular processes affected by CDK7 inhibition were analyzed by reverse transcriptase-quantitative PCR, western blot, flow cytometry and immunofluorescence. THZ1 perturbed the transcriptome and disabled CDK activation, leading to cell cycle arrest at G2 and DNA damage. THZ1 halted transcription of the nuclear-encoded mitochondrial ribosomal genes, reducing mitochondrial translation and oxidative respiration. It also inhibited the expression of receptor tyrosine kinases such as epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor-α (PDGFR-α), reducing signaling flux through the AKT, extracellular-signal-regulated kinase 1/2 (ERK1/2) and signal transducer and activator of transcription 3 (STAT3) downstream pathways. Finally, THZ1 disrupted nucleolar, Cajal body and nuclear speckle formation, resulting in reduced cytosolic translation and malfunction of the spliceosome and thus leading to aberrant mRNA processing. These findings indicate that CDK7 is crucial for gliomagenesis, validate CDK7 as a therapeutic target and provide new insight into the cellular processes that are affected by THZ1 and induce antitumor activity.

Sorption of selected endocrine disrupting chemicals to different aquatic colloids.

The sorption of seven endocrine disrupting chemicals (EDCs) to aquatic colloids was determined by cross-flow ultrafiltration (CFUF) followed by gas chromatography-mass spectrometry (GC-MS). Results show that the colloidal organic carbon normalized sorption coefficient (Kcoc) of EDCs to different aquatic colloids varies by a factor of 6-12 because such colloids are of different origin. Through characterization of colloidal samples, a significant relationship was established between Kcoc values and the molar extinction coefficient of colloids at 280 nm, whereas no other colloidal properties such as elemental ratios were correlated with Kcoc values. The results are consistent with other reports of the importance of the quality of sorbents such as their aromatic carbon content in sorbing various organic pollutants. The presence of a surfactant was found to increase Kcoc values for estrone (El) and 17alpha-ethynylestradiol (EE2). The method was subsequently applied for determining EDC concentrations in field samples, where both conventional and truly dissolved EDCs showed higher concentrations close to sewage outfalls than either upstream or downstream, confirming the sourceconcentration relationship. In addition, the truly dissolved EDC concentrations were lower than the conventional dissolved concentrations, confirming that there were interactions between aquatic colloids and EDCs. It is estimated that between 10 and 29% of EDCs are associated with aquatic colloids. As colloids are highly abundant in rivers and ocean, they will therefore play a significant role in the environmental behavior and fate of EDCs.

Microwave-assisted extraction followed by gas chromatography-mass spectrometry for the determination of endocrine disrupting chemicals in river sediments.

In this study, microwave-assisted extraction (MAE) followed by gas chromatography (GC)-mass spectrometry (MS) analysis has been successfully developed for the simultaneous extraction and determination of contrasting endocrine disrupting chemicals (EDCs) including 17beta-estradiol, estrone, 17(alpha-ethynylestradiol, 16alpha-hydroxyestrone, 4-nonylphenol, 4-tert-octylphenol and bisphenol A in river sediments. For MAE, the effects of various parameters on the extraction efficiency were investigated. It is shown that the most efficient extraction (recovery > 74%) of the target compounds was achieved by using methanol as the solvent, an extraction temperature of 110 degrees C and 15 min of holding time. The cleanup of extracts was carried out by passage through a non-deactivated silica gel column, and a satisfactory elution efficiency of all compounds was achieved using a solvent mixture of ethyl acetate-hexane (4:6, v/v). The spiking experiments show that the mean recovery of the target compounds exceeded 61% at a spiking level of 5 ng/g dry mass, and 73% at 10, 40 and 100 ng/g dry mass with a good reproducibility. The method developed was applied to the determination of target EDCs in river sediments collected from rivers Uck and Ouse, UK, and results revealed the presence of the chosen compounds at low ng/g level.

Simultaneous determination of endocrine disrupting phenolic compounds and steroids in water by solid-phase extraction-gas chromatography-mass spectrometry.

A solid-phase extraction (SPE)-gas chromatography (GC)-mass spectrometry (MS) analytical method for the simultaneous separation and determination of endocrine disrupting chemicals (EDCs) from water samples is described in detail. Important and contrasting EDCs including estrone, 17beta-estradiol, 17beta-ethynylestradiol, 16beta-hydroxyestrone, 4-nonylphenol, bisphenol A and 4-tert-octylphenol were selected as the target compounds. The SPE technique, followed by the derivatisation with bis (trimethylsilyl) trifluoroacetamide was used for the extraction recoveries of target compounds from water samples. A number of parameters that may affect the recovery of EDCs, such as the type of SPE cartridges, eluents, as well as water properties including pH value, and concentration of salts and humic substances were investigated. It is shown that the Oasis cartridges produced the best recoveries of target EDCs while ethyl acetate was efficient in eluting EDCs from SPE cartridges. The recovery of some EDCs was enhanced by the addition of salt, but reduced by the increase in pH value and humic acid concentration. The optimised method was further verified by performing spiking experiments in natural river water and seawater matrices, with good recovery and reproducibility for all the selected compounds. The established method was successfully applied to environmental water samples from East and West Sussex, UK, for the determination of the target EDCs.