Functional comparison of human and murine equilibrative nucleobase transporter 1.

6-Mercaptopurine (6-MP) maintenance therapy is the mainstay for various types of leukemia and inflammatory bowel disease. 6-MP is associated with numerous adverse effects including gastrointestinal intolerance, myelotoxicity, and hepatotoxicity. This can lead to therapy discontinuation which is associated with a higher risk of relapse. Drug transporter expression is a known factor contributing to patient variability in drug response and toxicity. We have established that the SLC43A3-encoded equilibrative nucleobase transporter 1 (ENBT1) mediates the transport of 6-MP into human lymphocytes and human embryonic kidney 293 (HEK293) cell lines transfected with SLC43A3. ENBT1 is known to be expressed in the gastrointestinal tract, bone marrow, and the liver. However, the relationship between ENBT1 and 6-MP-associated adverse events, and its pharmacokinetics, is unknown. To validate the use of mouse models (e.g. slc43a3-null mice) for exploring this relationship, we assessed the functional similarities between human and murine ENBT1 using HEK293 cells transfected with the respective SLC43A3/slc43a3 constructs, and the leukemia cell lines MOLT-4 (human) and L1210 (murine). Based on in silico analyses of structural similarities between transporters, we hypothesized that human and murine ENBT1 will have similar 6-MP transport/inhibition kinetics and a similar impact on 6-MP-induced cytotoxicity. We show herein that mslc43a3-encoded mouse ENBT1 transports both [3H]6-MP and [3H]adenine with kinetics similar to those of hSLC43A3-encoded human ENBT1. Both are also similarly distributed in mouse and human tissues. Therefore, data obtained from mouse models where ENBT1 is disrupted or modified may provide clinically relevant insights on its roles in modulating the actions of 6-MP.

The genome sequence of the Spruce-seed moth, Cydia strobilella (Linnaeus, 1758).

We present a genome assembly from an individual male Cydia strobilella (the spruce-seed moth; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 542.6 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.78 kilobases in length. Gene annotation of this assembly on Ensembl identified 13,058 protein coding genes.

Development of a Novel HEK293 Cell Model Lacking SLC29A1 to Study the Pharmacology of Endogenous SLC29A2-Encoded Equilibrative Nucleoside Transporter Subtype 2.

Equilibrative nucleoside transporters (ENTs) mediate the transmembrane flux of endogenous nucleosides and nucleoside analogs used clinically. The predominant subtype, ENT1, has been well characterized. However, the other subtype, ENT2, has been less well characterized in its native milieu due to its relatively low expression and the confounding influence of coexpressed ENT1. We created a cell model where ENT1 was removed from human embryonic kidney (HEK293) cells using CRISPR/cas9 [ENT1 knockout (KO) cells]; this cell line has ENT2 as the only functional purine transporter. Transporter function was assessed through measurement of [3H]2-chloroadenosine uptake. ENT1 protein was quantified based on the binding of [3H]nitrobenzylthioinosine, and ENT1/ENT2 protein was detected by immunoblotting. Changes in expression of relevant transporters and enzymes involved in purine metabolism were examined by quantitative polymerase chain reaction. Wild-type HEK293 cells and ENT1KO cells had a similar expression of SLC29A2/ENT2 transcript/protein and ENT2-mediated [3H]2-chloroadenosine transport activity (Vmax values of 1.02 ± 0.06 and 1.50 ± 0.22 pmol/µl/s, respectively). Of the endogenous nucleosides/nucleobases tested, adenosine had the highest affinity (Ki) for ENT2 (2.6 µM), while hypoxanthine was the only nucleobase with a submillimolar affinity (320 µM). A range of nucleoside/nucleobase analogs were also tested for their affinity for ENT2 in this model, with affinities (Ki) ranging from 8.6 µM for ticagrelor to 2,300 µM for 6-mercaptopurine. Our data suggest that the removal of endogenous ENT1 from these cells does not change the expression or function of ENT2. This cell line should prove useful for the analysis of novel drugs acting via ENT2 and to study ENT2 regulation. SIGNIFICANCE STATEMENT: We have created a cell line whereby endogenous ENT2 can be studied in detail in the absence of the confounding influence of ENT1. Loss of ENT1 has no impact on the expression and function of ENT2. This novel cell line will provide an ideal model for studying drug interactions with ENT2 as well as the cellular regulation of ENT2 expression and function.

The genome sequence of the Gold Triangle, Hypsopygia costalis (Fabricius, 1775).

We present a genome assembly from an individual male Hypsopygia costalis (the Gold Triangle; Arthropoda; Insecta; Lepidoptera; Pyralidae). The genome sequence is 818 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules with the Z sex chromosome assembled. The mitochondrial genome has also been assembled and is 15.3 kilobases in length. Gene annotation of this assembly on Ensembl identified 19,248 protein coding genes.

The Concise Guide to PHARMACOLOGY 2023/24: Transporters.

The Concise Guide to PHARMACOLOGY 2023/24 is the sixth in this series of biennial publications. The Concise Guide provides concise overviews, mostly in tabular format, of the key properties of approximately 1800 drug targets, and over 6000 interactions with about 3900 ligands. There is an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands (https://www.guidetopharmacology.org/), which provides more detailed views of target and ligand properties. Although the Concise Guide constitutes almost 500 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point-in-time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.16182. Transporters are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein-coupled receptors, ion channels, nuclear hormone receptors, catalytic receptors and enzymes. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid-2023, and supersedes data presented in the 2021/22, 2019/20, 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the Nomenclature and Standards Committee of the International Union of Basic and Clinical Pharmacology (NC-IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.

A SACS deletion variant in Great Pyrenees dogs causes autosomal recessive neuronal degeneration.

ARSACS (autosomal recessive spastic ataxia of Charlevoix-Saguenay) is a human neurological disorder characterized by progressive cerebellar ataxia and peripheral neuropathy. A recently recognized disorder in Great Pyrenees dogs is similarly characterized by widespread central nervous system degeneration leading to progressive cerebellar ataxia and spasticity, combined with peripheral neuropathy. Onset of clinical signs occurred in puppies as young as 4 months of age, with slow progression over several years. A multi-generation pedigree suggested an autosomal recessive mode of inheritance. Histopathology revealed consistent cerebellar Purkinje cell degeneration, neuronal degeneration in brainstem nuclei, widespread spinal cord white matter degeneration, ganglion cell degeneration, inappropriately thin myelin sheaths or fully demyelinated peripheral nerve fibers, and normal or only mild patterns of denervation atrophy in skeletal muscles. Genome-wide single nucleotide polymorphism (SNP) genotype data was collected from 6 cases and 26 controls, where homozygosity mapping identified a 3.3 Mb region on CFA25 in which all cases were homozygous and all controls were either heterozygous or homozygous for alternate haplotypes. This region tagged the SACS gene where variants are known to cause ARSACS. Sanger sequencing of SACS in affected dogs identified a 4 bp deletion that causes a frame shift and truncates 343 amino acids from the C terminus of the encoded sacsin protein (p.Val4244AlafsTer32). Our clinical and histopathological descriptions of this canine disorder contribute to the description of human ARSACS and represents the first naturally occurring large animal model of this disorder.

The genome sequence of the White-shouldered Marble, Apotomis turbidana (Hübner, 1825).

We present a genome assembly from an individual male Apotomis turbidana (the White-shouldered Marble; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 720.5 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 16.8 kilobases in length. Gene annotation of this assembly on Ensembl identified 22,646 protein coding genes.

The genome sequence of the White-barred Gold, Micropterix aruncella (Scopoli, 1763).

We present a genome assembly from an individual female Micropterix aruncella (the White-barred Gold; Arthropoda, Insecta, Lepidoptera; Micropterigidae). The genome sequence is 1,079 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 15.0 kilobases in length.

The genome sequence of the Elbow-stripe Grass-veneer, Agriphila geniculea (Haworth, 1811).

We present a genome assembly from an individual female Agriphila geniculea (the Elbow-stripe Grass-veneer; Arthropoda; Insecta; Lepidoptera; Crambidae). The genome sequence is 781.6 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z and W sex chromosomes. The mitochondrial genome has also been assembled and is 15.4 kilobases in length. Gene annotation of this assembly on Ensembl identified 22,132 protein coding genes.

Poverty dynamics and the determining factors among East African smallholder farmers.

CONTEXT: Rapid economic development in East Africa is matched by extremely dynamic smallholder livelihoods. Objective: To quantify the changes in poverty of smallholder farmers, to evaluate the potential of farm and off-farm activities to alleviate poverty, and to evaluate the potential barriers to poverty alleviation. METHODS: The analyses were based on a panel survey of 600 households undertaken in 2012 and re-visited approximately four years later in four sites in East Africa. The sites represented contrasting smallholder farming systems, linked to urban centres undergoing rapid economic and social change (Nairobi, Kampala, Kisumu, and Dar-es-Salaam). The surveys assessed farm management, farm productivity, livelihoods, and various measures of household welfare. RESULTS AND CONCLUSIONS: Almost two thirds of households rose above or fell below meaningful poverty thresholds - more than previously measured in this context - but overall poverty rates remained constant. Enhanced farm value production and off-farm income proved to be important mechanisms to rise out of poverty for households that were already resource-endowed. However, households in the poorest stratum in both panels appeared to be stuck in a poverty trap. They owned significantly fewer productive assets in the first panel compared to other groups (land and livestock), and these baseline assets were found to be positively correlated with farm income in the second panel survey. Equally these households were also found to be among the least educated, while education was found to be an important enabling factor for the generation of high value off-farm income. SIGNIFICANCE: Rural development that aims to stimulate increases in farm produce value as a means to alleviate poverty are only viable for already resource-endowed households, as they have the capacity to enhance farm value production. Conversely, the alleviation of extreme poverty should focus on different means, perhaps cash transfers, or the development of more sophisticated social safety nets. Furthermore, while off-farm income presents another important mechanism for poverty alleviation in rural areas, these opportunities are restricted to those households that have had access to education. As more households turn to off-farm activities to supplement or replace their livelihoods, farming approaches will also change affecting the management of natural resources. These dynamics ought to be better understood to better manage land-use transitions.

A history of shoulder instability is more common in young patients undergoing total shoulder arthroplasty.

BACKGROUND: Previous studies have shown an association between shoulder instability and the development of glenohumeral arthritis leading to total shoulder arthroplasty (TSA). The primary goal of this study was to evaluate if a history of shoulder instability was more common in patients aged <50 years undergoing TSA. The secondary objective was to determine if a history of prior surgical stabilization is more common in patients aged <50 years undergoing TSA. METHODS: Using the military health system data repository (MDR) and the Military Analysis and Reporting Tool (M2), we identified 489 patients undergoing primary TSA from October 1, 2013, to May 1, 2020, within the Military Health System (MHS). Patients aged <50 years were matched 1:2 with patients aged ≥50 years based on sex, race, and military status, with the final study population comprising 240 patients who underwent primary TSA during the study period. Electronic medical records were examined, and factors showing univariate association (P < .2) were included in a binary logistic regression analysis to determine associations between demographic or clinical factors and TSA prior to age 50 years. RESULTS: The groups differed significantly in shoulder arthritis subtype, with the older group having significantly more primary osteoarthritis (78% vs. 51%, P < .001). The younger group had significantly more patients with a history of shoulder instability (48% vs. 12%, P < .001), prior ipsilateral shoulder surgery of any type (74% vs. 34%, P < .001), and prior ipsilateral shoulder stabilization surgery (31% vs. 5%, P < .001). In the resultant logistic regression model, a history of shoulder instability (OR 5.0, P < .001) and a history of any prior ipsilateral shoulder surgery (OR 3.5, P < .001) were associated with TSA prior to the age of 50 years. CONCLUSIONS: Shoulder instability is a risk factor for TSA before age 50 years. It is unclear how surgical stabilization influences the development of secondary glenohumeral arthritis in shoulder instability. Patients should be counseled that recurrent instability could lead to earlier TSA, regardless of whether surgical stabilization is performed.

The genome sequence of the Sycamore Piercer, Pammene aurita (Razowski, 1991).

We present a genome assembly from an individual male Pammene aurita (the Sycamore Piercer; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 1,041.8 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 16.7 kilobases in length.

The genome sequence of the Ruddy Flat-body, Agonopterix subpropinquella (Stainton, 1849).

We present a genome assembly from an individual male Agonopterix subpropinquella (the Ruddy Flat-body; Arthropoda; Insecta; Lepidoptera; Depressariidae). The genome sequence is 667.9 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.5 kilobases in length. Gene annotation of this assembly on Ensembl identified 18,796 protein coding genes.

The genome sequence of the Notch-wing Button, Acleris emargana (Fabricius, 1775).

We present a genome assembly from an individual male Acleris emargana (the Notch-wing Button; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 691.4 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.34 kilobases in length. Gene annotation of this assembly on Ensembl identified 21,886 protein coding genes.

The genome sequence of the Lichen Button, Acleris literana (Linnaeus, 1758).

We present a genome assembly from an individual male Acleris literana (the Lichen Button; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 674.9 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.4 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,577 protein coding genes.

The genome sequence of the Heath Knot-horn, Apomyelois bistriatella (Hulst, 1887).

We present a genome assembly from an individual female Apomyelois bistriatella (the Heath Knot-horn; Arthropoda; Insecta; Lepidoptera; Pyralidae). The genome sequence is 389.6 megabases in span. Most of the assembly is scaffolded into 32 chromosomal pseudomolecules, including the Z and W sex chromosomes. The mitochondrial genome has also been assembled and is 15.2 kilobases in length.

The genome sequence of the Diamond-back Marble, Eudemis profundana (Denis & Schiffermüller, 1775).

We present a genome assembly from an individual male Eudemis profundana (the Diamond-back Marble; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 691.3 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.5 kilobases in length.

The genome sequence of the Ashy Button, Acleris sparsana (Denis & Schiffermüller, 1775).

We present a genome assembly from an individual male Acleris sparsana (the Ashy Button; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 589.5 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 16.4 kilobases in length. Gene annotation of this assembly on Ensembl identified 22,123 protein coding genes.

The genome sequence of the Thicket Knot-horn, Acrobasis suavella (Zincken, 1818).

We present a genome assembly from an individual male Acrobasis suavella (the Thicket Knot-horn; Arthropoda; Insecta; Lepidoptera; Pyralidae). The genome sequence is 647.3 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.31 kilobases in length. Gene annotation of this assembly on Ensembl identified 19,101 protein coding genes.

The genome sequence of the Grey Poplar Bell, Epinotia nisella (Clerck, 1759).

We present a genome assembly from an individual male Epinotia nisella (the Grey Poplar Bell; Arthropoda; Insecta; Lepidoptera; Tortricidae). The genome sequence is 585.0 megabases in span. Most of the assembly is scaffolded into 28 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.44 kilobases in length. Gene annotation of this assembly on Ensembl identified 18,952 protein coding genes.