R-Spondin1 enhances wnt signaling and decreases weight loss in short bowel syndrome zebrafish

BackgroundR-spondins, including R-spondin 1 (RSPO1), are a family of Wnt ligands that help to activate the canonical Wnt/β-catenin pathway, which is critical for intestinal epithelial cell proliferation and maintenance of intestinal stem cells. This proliferation underpins the epithelial expansion, or intestinal adaptation (IA), that occurs following massive bowel resection and short bowel syndrome (SBS). The purpose of this study was to identify if recombinant human RSPO1 (rhRSPO1) could be serially administered to SBS zebrafish to enhance cellular proliferation and IA.MethodsAdult male zebrafish were assigned to four groups: sham + PBS, SBS + PBS, sham + rhRSPO1, and SBS + rhRSPO1. Sham fish had a laparotomy alone. SBS fish had a laparotomy with distal intestinal ligation and creation of a proximal stoma. Fish were weighed at initial surgery and then weekly. rhRSPO1 was administered post-operatively following either a one- or two-week dosing schedule with either 3 or 5 intraperitoneal injections, respectively. Fish were harvested at 7 or 14 days with intestinal segments collected for analysis.ResultsRepeated intraperitoneal injection of rhRSPO1 was feasible and well tolerated. At 7 days, intestinal epithelial proliferation was increased by rhRSPO1. At 14 days, SBS + rhRSPO1 fish lost significantly less weight than SBS + PBS fish. Measurements of intestinal surface area were not increased by rhRSPO1 administration but immunofluorescent staining for β-catenin and gene expression for cyclin D1 was increased.ConclusionsIntraperitoneal injection of rhRSPO1 decreased weight loss in SBS zebrafish with increased β-catenin + cells and cyclin D1 expression at 14 days, indicating improved weight maintenance might result from increased activation of the canonical Wnt pathway.     

Studies in the genus Hypericum L. (Clusiaceae). 1 Section 9. Hypericum sensu lato (part 3): Subsection 1. Hypericum series 2. Senanensia,subsection 2. Erecta and section 9b. Graveolentia

Abstract

Part 4(3) of this monographic series of papers on the genus Hypericum is prefaced by an introduction to the genus and a summary of the aims and methods of the project. This is followed by treatments of the remaining parts of sect. 9. Hypericum sensu stricto and the last segregate section from the original sect. Hypericum, sect. 9b. Graveolentia. Both hitherto untreated parts of the reduced sect. Hypericum are mainly Japanese, but some species extend in distribution as far as Kamchatka, eastern Siberia, central China, and Sabah (Mt. Kinabalu). Sect. Graveolentia is North and Central American. Sect. Hypericum subsect. Hypericum series Senanensia contains seven species from northern Japan and adjacent areas, including H. pibairense (Miyabe & Y. Kimura) N. Robson, stat. nov., H. nakaii subsp. miyabei (Y. Kimura) N. Robson, comb. et stat. nov., H. nakaii subsp. tatewakii (S. Watanabe) N. Robson, comb. et stat. nov. and H. senanense subsp. mutiloides (R. Keller) N. Robson, comb. et stat. nov. Sect. Hypericum subsect. Erecta contains 23 species and one hybrid from Japan, Korea, central China, Taiwan, Luzon, Sabah and Sumatera, including H. kawaranum N. Robson, stat. et nom. nov., H. watanabei N. Robson, stat. et nom. nov., H. kimurae N. Robson, stat. et nom. nov., H. pseudoerectum stat. et nom. nov., H. kitamense (Y. Kimura) N. Robson, stat. nov., H. kurodakeanum N. Robson, stat. et nom. nov., H. furusei N. Robson, sp. nov., H. nuporoense N. Robson, sp. nov. and H. ovalifolium subsp. hisauchii (Y. Kimura) N. Robson, stat. nov. Sect. Graveolentia contains nine species and one hybrid from southeastern Canada, the eastern half of the United States, Mexico and western Guatemala, including H. oaxacanum subsp. veracrucense N. Robson, subsp. nov. and H. macvaughii N. Robson, sp. nov.     

Synthetic Studies on Formamidopyrimidines Related to Clofarabine

Degradation of clofarabine (3) in 0.9% saline solution at 100°C afforded three degradation products which were determined to be formamidopyrimidines 4–6.Compounds 4 and 5 were assigned as C_1′ anomers on the basis of one-dimensional and two-dimensional NMR experiments, whereas 6 was found to be the formamidopyrimidine lacking the sugar moiety. An improved procedure for the synthesis of formamidopyrimidines was developed, wherein benzoylated clofarabine (11) was treated with allyl chloroformate, followed by deprotection of the alloc group with catalytic Pd(PPh₃)₄ and dimedone. A synthesis of compound 6 from 4 is also described.     

Differences in acute stress responses between wild-caught and captive-bred birds: a physiological mechanism contributing to current avian invasions?

Current avian invasions are often the result of exotic birds accidentally escaping from cages. It has been hypothesised that the higher invasiveness of wild-caught cage-birds compared to captive-bred ones could be related to the loss of ability in captive-bred birds to cope with new environments. The acute stress response plays an important role in how animals cope with challenges because elevated corticosterone (CORT) levels can mediate learning and memory consolidation and help to increase their survival prospects. We experimentally tested whether exotic wild-caught and captive-bred cage-birds differ in their responses to acute stress using a representative sample of parrots. Wild-caught individuals showed longer CORT responses to acute stress than captive-bred ones, both at inter- and intra-specific levels when comparing wild-birds to the first generation born in captivity. Captive-bred birds may have attenuated their CORT responses due to acclimation, while high mortality rates during the international trade of wild-caught birds could have selected those individuals that are better able to cope with stress. We suggest that the longer acute response found in wild-caught parrots could help them to escape from cages and survive when facing challenges in new wild environments, possibly contributing to their higher invasiveness.     

Assessment of SLX4 Mutations in Hereditary Breast Cancers

Background

SLX4 encodes a DNA repair protein that regulates three structure-specific endonucleases and is necessary for resistance to DNA crosslinking agents, topoisomerase I and poly (ADP-ribose) polymerase (PARP) inhibitors. Recent studies have reported mutations in SLX4 in a new subtype of Fanconi anemia (FA), FA-P. Monoallelic defects in several FA genes are known to confer susceptibility to breast and ovarian cancers.

Methods and Results

To determine if SLX4 is involved in breast cancer susceptibility, we sequenced the entire SLX4 coding region in 738 (270 Jewish and 468 non-Jewish) breast cancer patients with 2 or more family members affected by breast cancer and no known BRCA1 or BRCA2 mutations. We found a novel nonsense (c.2469G>A, p.W823*) mutation in one patient. In addition, we also found 51 missense variants [13 novel, 23 rare (MAF<0.1%), and 15 common (MAF>1%)], of which 22 (5 novel and 17 rare) were predicted to be damaging by Polyphen2 (score = 0.65–1). We performed functional complementation studies using p.W823* and 5 SLX4 variants (4 novel and 1 rare) cDNAs in a human SLX4-null fibroblast cell line, RA3331. While wild type SLX4 and all the other variants fully rescued the sensitivity to mitomycin C (MMC), campthothecin (CPT), and PARP inhibitor (Olaparib) the p.W823* SLX4 mutant failed to do so.

Conclusion

Loss-of-function mutations in SLX4 may contribute to the development of breast cancer in very rare cases.     

Introduced populations as genetic reservoirs for imperiled species: a case study of the Arkansas River Shiner (Notropis girardi)

The Arkansas River Shiner is a threatened species that has been extirpated throughout much of its native range (Arkansas River drainage) and remaining populations are imperiled. Prior to 1978, this species was accidently introduced to the Pecos River (Rio Grande drainage) via bait bucket, and has since persisted for over 30 years. Genetic data show that the Pecos River population maintains comparable levels of diversity at mitochondrial DNA and microsatellite loci relative to native range populations. Hence, we examined several factors that could be responsible for high introduced genetic diversity including (a) multiple introductions from genetically distinct sources (b) introduction of individuals from a genetically diverse source followed by rapid population expansion, (c) presence of life-history traits that foster propagule diversity and wide spatio-temporal demographic and genetic mixing; and (d) introduction to a suitable habitat in the non-native range. Our findings indicate Arkansas River Shiner was likely introduced from the Canadian River and subsequently experienced rapid population expansion that mitigated loss of diversity during the founding event. Threats to native Arkansas River Shiner have increased due to ongoing drought and water resource development, thus a finding of high diversity in the Pecos River suggests conservation significance of this non-native population. Further, it identifies the Pecos River as both a refuge for native endemic fishes and of genetic diversity of introduced, yet threatened, species.     

SLC26A4 Targeted to the Endolymphatic Sac Rescues Hearing and Balance in Slc26a4 Mutant Mice

Mutations of SLC26A4 are a common cause of human hearing loss associated with enlargement of the vestibular aqueduct. SLC26A4 encodes pendrin, an anion exchanger expressed in a variety of epithelial cells in the cochlea, the vestibular labyrinth and the endolymphatic sac. Slc26a4 ^Δ/Δ mice are devoid of pendrin and develop a severe enlargement of the membranous labyrinth, fail to acquire hearing and balance, and thereby provide a model for the human phenotype. Here, we generated a transgenic mouse line that expresses human SLC26A4 controlled by the promoter of ATP6V1B1. Crossing this transgene into the Slc26a4 ^Δ/Δ line restored protein expression of pendrin in the endolymphatic sac without inducing detectable expression in the cochlea or the vestibular sensory organs. The transgene prevented abnormal enlargement of the membranous labyrinth, restored a normal endocochlear potential, normal pH gradients between endolymph and perilymph in the cochlea, normal otoconia formation in the vestibular labyrinth and normal sensory functions of hearing and balance. Our study demonstrates that restoration of pendrin to the endolymphatic sac is sufficient to restore normal inner ear function. This finding in conjunction with our previous report that pendrin expression is required for embryonic development but not for the maintenance of hearing opens the prospect that a spatially and temporally limited therapy will restore normal hearing in human patients carrying a variety of mutations of SLC26A4.     

Effects of X-Ray Dose On Rhizosphere Studies Using X-Ray Computed Tomography

X-ray Computed Tomography (CT) is a non-destructive imaging technique originally designed for diagnostic medicine, which was adopted for rhizosphere and soil science applications in the early 1980s. X-ray CT enables researchers to simultaneously visualise and quantify the heterogeneous soil matrix of mineral grains, organic matter, air-filled pores and water-filled pores. Additionally, X-ray CT allows visualisation of plant roots in situ without the need for traditional invasive methods such as root washing. However, one routinely unreported aspect of X-ray CT is the potential effect of X-ray dose on the soil-borne microorganisms and plants in rhizosphere investigations. Here we aimed to i) highlight the need for more consistent reporting of X-ray CT parameters for dose to sample, ii) to provide an overview of previously reported impacts of X-rays on soil microorganisms and plant roots and iii) present new data investigating the response of plant roots and microbial communities to X-ray exposure. Fewer than 5% of the 126 publications included in the literature review contained sufficient information to calculate dose and only 2.4% of the publications explicitly state an estimate of dose received by each sample. We conducted a study involving rice roots growing in soil, observing no significant difference between the numbers of root tips, root volume and total root length in scanned versus unscanned samples. In parallel, a soil microbe experiment scanning samples over a total of 24 weeks observed no significant difference between the scanned and unscanned microbial biomass values. We conclude from the literature review and our own experiments that X-ray CT does not impact plant growth or soil microbial populations when employing a low level of dose (<30 Gy). However, the call for higher throughput X-ray CT means that doses that biological samples receive are likely to increase and thus should be closely monitored.