Multiple Origins and Genomic Basis of Complex Traits in Sighthounds.

Sighthounds, a distinctive group of hounds comprising numerous breeds, have their origins rooted in ancient artificial selection of dogs. In this study, we performed genome sequencing for 123 sighthounds, including one breed from Africa, six breeds from Europe, two breeds from Russia, and four breeds and 12 village dogs from the Middle East. We gathered public genome data of five sighthounds and 98 other dogs as well as 31 gray wolves to pinpoint the origin and genes influencing the morphology of the sighthound genome. Population genomic analysis suggested that sighthounds originated from native dogs independently and were comprehensively admixed among breeds, supporting the multiple origins hypothesis of sighthounds. An additional 67 published ancient wolf genomes were added for gene flow detection. Results showed dramatic admixture of ancient wolves in African sighthounds, even more than with modern wolves. Whole-genome scan analysis identified 17 positively selected genes (PSGs) in the African population, 27 PSGs in the European population, and 54 PSGs in the Middle Eastern population. None of the PSGs overlapped in the three populations. Pooled PSGs of the three populations were significantly enriched in "regulation of release of sequestered calcium ion into cytosol" (gene ontology: 0051279), which is related to blood circulation and heart contraction. In addition, ESR1, JAK2, ADRB1, PRKCE, and CAMK2D were under positive selection in all three selected groups. This suggests that different PSGs in the same pathway contributed to the similar phenotype of sighthounds. We identified an ESR1 mutation (chr1: g.42,177,149 T > C) in the transcription factor (TF) binding site of Stat5a and a JAK2 mutation (chr1: g.93,277,007 T > A) in the TF binding site of Sox5. Functional experiments confirmed that the ESR1 and JAK2 mutation reduced their expression. Our results provide new insights into the domestication history and genomic basis of sighthounds.

Unveiling fenpropathrin resistance levels in field populations of Tetranychus cinnabarinus (Boisduval): Insights, risks, and RNAi strategy.

Indoor cases of Tetranychus cinnabarinus displaying resistance have been documented, but the resistance level in field populations remains unexplored in China. This study delves into the resistance dynamics of T. cinnabarinus to fenpropathrin in various field populations across China, a pressing concern in contemporary agricultural pest control. The conventional bioassay and amplicon sequencing reveal a notable absence of significant fenpropathrin resistance in field populations, contrasting with known resistance in indoor cases. Current study highlights the limitations of traditional bioassays in detecting early-stage resistance and underscores the nuanced capabilities and constraints of amplicon sequencing in resistance gene frequency analysis. By employing an integrated approach, we combined dose-response bioassays, amplicon sequencing, and statistical modeling to assess resistance levels and investigate underlying genetic factors. The model with empirical data indicates that a 5% mutation frequency represents the threshold before resistance emerges. However, the detection of the kdr mutation in certain populations ranging from 0 to 1.2%, signals an early looming threat of future resistance emergence. Additionally, we further assessed a specific dsRNA targeting VGSC genes at two concentrations (10 ng/µL and 100 ng/µL), both inducing substantial mortality by silencing target genes effectively. The exploration of RNA interference (RNAi) as a novel, more environmentally friendly pest control measure opens new avenues, despite the ongoing challenge of resistance evolution. Overall, this study underscores the necessity for evolving pest management strategies, integrating advanced biotechnological approaches with traditional methods, to effectively counter pesticide resistance and ensure sustainable agricultural productivity.

[Evaluating the risk of ischemic cerebral vascular disease with ocular hemodynamics in carotid artery stenosis patients].

OBJECTIVE: To evaluate the association between the ocular hemodynamic and the incidence of ischemic cerebral vascular diseases in carotid artery stenosis patients. METHODS: Case-control study. Seventy-two cases patients with carotid artery stenosis including 22 cases of cerebral infarction group, 14 cases of lacunar cerebral infarction group and the control group of the other 36 patients without cerebral infarction were recruited. All patients were measured with hemodynamic parameters of the ophthalmic artery, central retinal artery and posterior ciliary artery. PSV, EDV, PI and RI were also measured. The other risk ischemic cerebral vascular factors were obtained through the questionnaire and venous blood sampling. The differences of 3 groups and the influences of the other risk factor of hemodynamic were calculated. RESULTS: PSV of ophthalmic artery was (25.96 ± 12.63)cm/s in cerebral infarction group, (32.63 ± 13.25) cm/s in lacunar infarction group and (39.46 ± 14.97) cm/s in control group. EDV of ophthalmic artery was(6.09 ± 3.10) cm/s in cerebral infarction group, (6.57 ± 2.85) cm/s in lacunar infarction group and (9.73 ± 5.02) cm/s in control group. PSV of central retinal artery was (8.06 ± 2.98) cm/s in cerebral infarction group, (8.91 ± 2.76) cm/s in lacunar infarction group and (9.71 ± 2.60) cm/s in control group. The differences were statistically significant (F = 13.393, 12.609, 5.027, P < 0.01). EDV of central retinal artery was (2.46 ± 0.77) cm/s in cerebral infarction group, (2.35 ± 0.89) cm/s in lacunar infarction group and (2.73 ± 0.86) cm/s in control group. There was no statistical difference of EDV of CRA in three group (F = 2.405, P = 0.094). CONCLUSIONS: There was association between the ocular hemodynamic parameters and the incidence of ischemic cerebrovascular disease in the carotid artery stenosis patients. Increased level of PSV and EDV of the ophthalmic artery raised the risk of the incidence of ischemic cerebrovascular disease.

Inhibition of glucocorticoid-induced alteration of vimentin by a glucocorticoid receptor antagonist RU486 in the organ-cultured rat lens.

PURPOSE: Long-term application of glucocorticoids as a treatment for conditions such as allergy, autoimmune diseases, and transplantation presents a high risk of development of steroid-induced cataract. The presence of a functional glucocorticoid receptor (GR) in human and rat lens epithelial cells suggests a direct and specific targeting of these lens cells by glucocorticoids. One important cytoskeletal protein in lens epithelial cells is vimentin, which plays an important role in maintaining the normal lens morphology and function. Previous studies have shown that vimentin is involved in signal transduction, changes in cell structure and differentiation, and apoptosis. Based on a model of steroid-induced cataract from our previous study, the present study focuses on whether changes in vimentin can be induced in vitro through specific GR activation in glucocorticoid-induced cataracts of the rat lens. METHODS: Clear rat lenses, cultured in vitro, were treated with or without dexamethasone (Dex) or RU486 (a glucocorticoid receptor antagonist). Lenses were cultured for 7 days at 37 °C under 5% CO2, and were observed daily with an inverted microscope. Changes in morphology were followed by Hematoxylin-eosin (HE) staining, transmission electron microscopy, and immunohistochemistry. The expression of vimentin mRNA and protein was examined by reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively, in the capsule-epithelium and fiber tissue of the lenses. RESULTS: Opacity was obviously present at day 7 in the Dex group. The lenses of the untreated group and the RU486+Dex group remained transparent throughout the incubation. Electron microscopy showed an orderly arrangement of fiber cells and normal cell junctions in the control group and the RU486+Dex group. However, in the Dex group, fiber cells were disarranged and the cell-cell junctions exhibited lacunae. The expression of vimentin protein in the lens capsule-epithelium and fiber tissue decreased in the Dex-treated group, but normal expression of vimentin mRNA was maintained. CONCLUSIONS: These results suggest that the GR-mediated reduction in vimentin may be involved in the formation of steroid-induced cataract.

Inhibition of glucocorticoid-induced changes of Na(+), K(+)-ATPase in rat lens by a glucocorticoid receptor antagonist RU486.

Cataract formation can be induced by prolonged use of glucocorticoids. The underlying mechanism is not fully understood yet. The presence of the functional glucocorticoid receptor (GR) in human and rat lens epithelial cells suggests that glucocorticoids target lens epithelial cells directly and specifically. Na(+), K(+)-ATPase has long been recognized for its role in regulating electrolyte concentration in the lens, contributing to lens transparency. We previously reported that the inactivation of Na(+), K(+)-ATPase induced by a glucocorticoid in rat lens. Therefore, the question is whether the changes of Na(+), K(+)-ATPase can be induced through the specific GR activation in glucocorticoid-induced cataract formation. Clear rat lenses were cultured in vitro and were treated with or without dexamethasone (Dex) or RU486 (a GR antagonist). The lenses were cultured for 7 days and photographed daily to record the development of opacity. The activity of Na(+), K(+)-ATPase was determined by using spectrophotometric analysis. The mRNA and protein level expressions of Na(+), K(+)-ATPase α1 were examined by reverse transcription polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry analysis, respectively. Our findings are presented in this study and show that mist-like opacity of the lens was observed as early as 5 days after incubation with dexamethasone. The opacity was more obvious at day 7 in the Dex group. The lenses of the untreated group and the RU486+Dex group remained transparent throughout the incubation. The activity of Na(+), K(+)-ATPase in the Dex-treated group decreased in a time-dependent manner. There was no significant loss of enzyme activity in either the control or the RU486+Dex group throughout the incubation period. Both the protein and mRNA expression levels of Na(+), K(+)-ATPase α1 in the capsule-epithelium of lenses decreased in the Dex-treated group. The GR antagonist RU486 inhibited the decrease of the expression of Na(+), K(+)-ATPase α1 induced by Dex. All of the above results suggested that the GR-mediated reduction of Na(+), K(+)-ATPase may contribute to the formation of steroid-induced cataract. Intervention in this pathway maybe helpful to avoid glucocorticoids-cataract formation.

BuyangHuanwu decoction (BYHWD) protects against retinal ischemic injury.

Severe ipsilateral or bilateral carotid artery stenosis or occlusion is the most common cause of ocular ischemic syndrome (OIS). This study aimed to evaluate the protective effects of BuyangHuanwu decoction (BYHWD) against the retinal ischemia induced by ligation of carotid arteries in rats. The effects of BYHWD (0.64 g/kg, i.g.) on ischemia-induced retinal damage were examined with histological staining, RT-PCR and western blot. The protective effects of BYHWD on the ischemia retinal ganglion cells (RGCs) model induced by hypoxia were examined in vitro. The results indicated that BYHWD significantly prevented the decrease in density of cells in ganglion cell layer (GCL) and in the thickness of total retinal, and the increase in the numbers of TUNEL-positive cells. In addition, BYHWD decreased the expression of Caspase-3, Caspase-8 and Bax, and increased the expression of Bcl-2 in retina after ischemia. Similar to the results in vivo, BYHWD significantly suppressed the hypoxia-induced changes of apoptosis related gene and protein expression. These findings suggest that BYHWD may prevent ischemia-induced retinal ischemia through its inhibition of apoptosis.

Collapsin Response Mediator Protein-2-induced Retinal Ischemic Injury in a Novel Mice Model of Ocular Ischemia Syndrome.

BACKGROUND: Collapsin response mediator protein-2 (CRMP2) has been shown to be involved in ischemia/hypoxia (IH) injury. We determined whether CRMP2 modulates ischemic injury in the retinal of Ocular ischemic syndrome (OIS). This study was to explore the molecular mechanisms underlying OIS in a novel mice model. METHODS: Experiments were performed on adult male C57/BL6 mice that received bilateral internal carotid arteries ligation for 1, 2, or 4 weeks. The mice received injection of calpeptin group before occlusion for 4 weeks or not. The expression of CRMP2 in the retinal was examined by western blotting (WB) analysis and immunohistochemical analysis (IHC). The effects of ischemic injury on retinal were evaluated by fundus examination, fundus fluorescein angiography, electroretinogram, cell counting of retinal ganglion cell (RGC), and measurement of the thickness of the retina. RESULTS: The veins dilated after chronic ischemia. In the electroretinography, the amplitudes of a- and b-waves kept diminishing in an ischemia time-dependent manner. Moreover, the tail vein-retinal circulation time prolonged in the 1- and 2-week group. In comparison, thickness of the retina decreased gradually with the ischemia time elapsed. WB analysis showed the CRMP2 and p-CRMP2 levels decreased in the 2- and 4-week groups. The results of IHC analysis were compatible with our results of WB. The loss of RGCs, decrease of the total reaction time and reduction of CRMP2 was alleviated by intravitreal injection of calpeptin. CONCLUSIONS: These results revealed that bilateral ligation of the internal carotid artery causes retinal ischemia in mice. Moreover, CRMP2 might play a pivotal role during the ischemic injury in the retina and inhibit the cleavage of CRMP2 can ameliorate the IH injury.