Should biochemical markers of bone turnover be considered standard practice for safety pharmacology?

The success in biomedical sciences such as genomics and proteomics is not paralleled in the medical product development methods. The consequence of this is a lack of translation into improved drug safety and efficacy. Therefore the US Food and Drug Administration (FDA) introduced the Critical Path Initiative in 2004 to modernize drug development and safety pharmacology. Bone is that largest tissue by weight, and is continuously remodelled. Changes in bone turnover lead to complications such as osteoporosis and fracture, that is associated with an increased mortality. Recent findings have identified bone as a possible endocrine organ and the availability of valid biochemical bone markers suggests that assessing bone turnover should also play an important role in general safety pharmacology.     

A combination of genome-wide association study and selection signature analysis dissects the genetic architecture underlying bone traits in chickens

The bones of chicken play an important role in supporting and protecting the body. The growth and development of bones have a substantial influence on the health and production performance in chickens. However, genetic architecture underlying chicken bone traits is not well understood. The objectives of this study are to dissect the genetic basis of bone traits in chickens and to identify valuable genes and genetic markers for chicken breeding. We performed a combination of genome-wide association study (GWAS) and selection signature analysis (fixation index values and nucleotide diversity ratios) in an F₂ crossbred experimental population with different genetic backgrounds (broiler × layer) to identify candidate genes and significant variants related to femur, shank, keel length, chest width, metatarsal claw weight, metatarsal length, and metatarsal circumference. A total of 545 individuals were genotyped based on the whole genome re-sequencing method (26 F₀ individuals were re-sequenced at 10 × coverage; 519 F₂ individuals were re-sequenced at 3 × coverage). A total of 2 028 112 single-nucleotide polymorphisms (SNPs) remained to carry out analysis after quality control and imputation. The integration of GWAS and selection signature analysis indicated that all significant SNPs responsible for bone traits were mainly localized on chicken chromosomes 1, 4, and 27. Finally, we identified 21 positional candidate genes that might regulate chicken bone growth and development, including LRCH1, RB1, FNDC3A, MLNR, CAB39L, FOXO1, LHFP, TRPC4, POSTN, SMAD9, RBPJ, PPARGC1A, SLIT2, NCAPG, NKX3-2, CPZ, SPOP, NGFR, SOST, ZNF652, and HOXB3. Additionally, an array of uncharacterized genes was identified. The findings provide an in-depth understanding of the genetic architecture of chicken bone traits and offer a molecular basis for applying genomics in practical chicken breeding.     

A factor synthesized by rabbit periosteal fibroblasts stimulates bone resorption and collagenase production by connective tissue cells in vitro

Unstimulated monolayer cultures of confluent rabbit periosteal fibroblasts synthesize a factor that stimulates bone resorption in vitro. Furthermore it stimulates rabbit chondrocytes and mouse osteoblasts to synthesize collagenase. The factor has no effect on dead bone in culture, and its activity on live bone is mediated principally by osteoclasts, since it is 75% inhibited by salmon calcitonin. Characterization of the factor by gel filtration and isoelectric focusing indicates an M_r in the range 15 000–25 000 and a pI corresponding to approx. pH 4.7. These biological and physicochemical properties are similar to those reported for a factor released by peripheral blood monocytes. However, whereas human monocyte factor in both the crude and partially-purified state exhibits interleukin-1 activity, crude and fractionated periosteal fibroblast-conditioned medium does not. This is the first report of a conditioned medium containing a molecule like the monocyte-factor which appears to have no interleukin 1 activity. The factor may be synthesized by a wide range of cell types, and could have an important role in mediating connective tissue degradation during both physiological and pathological resorption.     

The synergistic role of Pu.1 and Fms in zebrafish osteoclast-reducing osteopetrosis and possible therapeutic strategies

Osteoclasts are bone resorption cells of myeloid origin. Osteoclast defects can lead to osteopetrosis, a genetic disorder characterized by bone sclerosis for which there is no effective drug treatment. It is known that Pu.1 and Fms are key regulators in myelopoiesis, and their defects in mice can lead to reduced osteoclast numbers and consequent osteopetrosis. Yet how Pu.1 and Fms genetically interact in the development of osteoclasts and the pathogenesis of osteopetrosis is still unclear. Here, we characterized pu.1^G242D;fms^j4e1 double-deficient zebrafish, which exhibited a greater deficiency of functional osteoclasts and displayed more severe osteopetrotic symptoms than the pu.1^G242D or fms^j4e1 single mutants, suggesting a synergistic function of Pu.1 and Fms in the regulation of osteoclast development. We further demonstrated that Pu.1 plays a dominant role in osteoclastogenesis, whereas Fms plays a dominant role in osteoclast maturation. Importantly, treatment with the drug retinoic acid significantly relieved the different degrees of osteopetrosis symptoms in these models by increasing the number of functional osteoclasts. Thus, we report the development of valuable animal models of osteopetrosis, and our results shed light on drug development for antiosteopetrosis therapy.     

Soil seed bank in different vegetation types in the Loess Plateau region and its role in vegetation restoration

In the Loess Plateau region, soil erosion is a serious problem. Vegetation restoration is an effective approach to control soil erosion and improve ecosystems. The soil seed bank generally plays an important role in vegetation restoration after disturbance. Thus, we reviewed soil seed bank studies to reveal the soil seed bank characteristics and its role in vegetation restoration in three vegetation types (forest, forest‐steppe, and steppe). We selected 38 seed bank studies and analyzed several seed bank characteristics, such as seed density, species composition, and the relationship between seed size and seed bank. We also assessed the role of the soil seed bank in vegetation restoration. The soil seed bank density ranged from 2,331 ± 1,993 to 6,985 ± 4,047 seeds/m² among the different vegetation types. In the soil seed bank, perennial herbs and grasses accounted for 51.5% of the total species. Native species that were dominant or common in the standing vegetation usually had relatively high seed bank densities. Moreover, species with smaller seeds generally had higher soil seed bank densities. The present study indicates that the soil seed bank plays a significant role in spontaneous vegetation restoration, especially during the early successional stages in abandoned slope farmlands and grazing‐excluded grasslands. However, species with large seeds or transient soil seed banks should be reintroduced through seeding to accelerate target species restoration. More studies on soil seed banks need to be conducted to comprehensively reveal their characteristics.     

Effects of manganese-supplemented diets on growth performance,blood biochemistry,nitrogen metabolism and skeletal development of rex rabbits

(Background) Manganese (Mn) is an essential mineral, although its effects on rabbits is not clear. (Research Purpose) This study was conducted to investigate the effects of the level of supplementation of dietary manganese on growth performance, blood biochemistry, nitrogen metabolism and skeletal development of growing Rex rabbits. (Methods) Two hundred 3-month-old healthy Rex rabbits with similar body weights were randomly divided into 5 groups (A, B, C, D, E), with 40 replicates in each group. The rabbits in the 5 groups were fed a basal diet containing 0, 5, 10, 20 and 40 mg/kg manganese (in the form of manganese sulfate), respectively. The trial included 7 days for adaptation and 29 days of testing. Seven days before the end of feeding, eight rabbits from each group were transferred into a metabolic cage for metabolic testing. (Results) The results showed that supplemental dietary manganese levels did not significantly influence final body weight (FBW) or average daily feed intake (ADFI) (P＞0.05). Average daily gains (ADG) were significantly higher in the 20 mg/kg manganese group than in the other groups, and the ratio of feed to body weight gain (F/G) was significantly affected by manganese level (P < 0.05). No significant differences were found in the digestion coefficients among the groups (P > 0.05). Regarding carcass traits, the thymus index and total fat were significantly different (P < 0.05) among the groups, but there were no other significant differences (P > 0.05). The addition of manganese had no significant effect on the intake of nitrogen (IN), fecal nitrogen (FN), digestible nitrogen (DN) or the apparent digestibility of nitrogen (NAD). Compared to the other groups, urinary nitrogen (UN) was lower in the 20 mg/kg group, although nitrogen deposition (RN), nitrogen utilization (NUR) and the biological potency of nitrogen (NBV) were higher in this group (P < 0.05). As the amount of manganese added to the diet increased, serum triglycerides decreased (P < 0.05). Serum Mn-SOD was significantly lower in the 5 mg/kg manganese group than in the other groups (P < 0.05). The results of this study demonstrate that a diet with supplemented manganese can improve Rex rabbit growth performance and increase RN, NUR and NBV. There were no significant effects of different dietary levels of Mn on the ratio of bone to meat (P > 0.05) or bone strength (P < 0.05). (Conclusion) In conclusion, we determined that the optimal level of manganese supplementation in the diet of growing Rex rabbits was 20 mg/kg, which was also found to reduce nitrogen emissions into the environment.     

黄土区土质与土石质塿土堆积体水力侵蚀过程差异

利用室内模拟降雨试验,研究了不同雨强及坡度条件下黄土区土质(不含砾石)与土石质(砾石质量分数30%)塿土堆积体的水动力学特征、侵蚀特征及侵蚀动力机制的差异。结果表明: 砾石存在改变了堆积体坡面的水动力学特性,与土质坡面相比,土石质坡面的流速、弗汝德数、单位径流功率和过水断面单位能分别减少1.7%～49.7%、6.7%～60.6%、2.0%～44.6%和1.0%～26.7%;曼宁糙率系数、径流剪切力分别增加6.2%～169.4%、5.7%～79.3%。2.0、2.5 mm·min^-1雨强下,土石质坡面侵蚀速率较土质坡面降低26.2%～89.9%,砾石的减沙效益显著。2种堆积体的侵蚀速率与水动力学参数间均可用线性函数拟合,与土质坡面相比,土石质坡面的可蚀性参数均降低,降幅为56.1%～73.3%;而临界水动力学参数中径流剪切力增加11.1%,径流功率、单位径流功率和过水断面单位能分别减少25.4%、64.0%和5.0%。砾石的存在一定程度上控制了工程堆积体坡面降雨侵蚀过程。     

宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性

目的探讨宫颈病变患者阴道微生态与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。方法选择2018年1月至2019年1月间在我院确诊为原发性宫颈癌的患者50例作为宫颈癌组,在我院诊断为宫颈糜烂的患者78例作为宫颈糜烂组,同期在我院进行体检的健康女性100例作为正常对照组。对比3组研究对象的阴道微生态失调率、高危型HPV感染率以及宫颈癌组、宫颈糜烂组患者宫颈病灶组织中宫颈癌相关增殖基因(Prdx4、Nek2、Fhit、BLCAP)mRNA表达量的差异。采用Pearson检验分析宫颈癌患者阴道微生态失调率与高危型HPV感染及宫颈癌相关增殖基因表达的相关性。结果宫颈癌组、宫颈糜烂组患者的阴道微生态失调率、高危型HPV感染率高于正常对照组,其中宫颈癌组患者这两项指标水平高于宫颈糜烂组(均P0.05)。宫颈癌组患者宫颈病灶组织中Prdx4、Nek2 mRNA表达量高于宫颈糜烂组,Fhit、BLCAP mRNA表达量低于宫颈糜烂组(均P0.05)。相关性分析发现,宫颈癌患者阴道微生态失调率与高危型HPV感染率呈正相关,与癌基因(Prdx4、Nek2)mRNA表达量呈正相关,与抑癌基因(Fhit、BLCAP)mRNA表达量呈负相关(均P0.05)。结论宫颈癌患者阴道微生态失调率较高,可能与高危型HPV感染及癌细胞增殖旺盛密切相关。     

Using the otolith sulcus to aid in prey identification and improve estimates of prey size in diet studies of a piscivorous predator

Diet studies are fundamental for understanding trophic connections in marine ecosystems. In the southeastern US, the common bottlenose dolphin Tursiops truncatus is the predominant marine mammal in coastal waters, but its role as a top predator has received little attention. Diet studies of piscivorous predators, like bottlenose dolphins, start with assessing prey otoliths recovered from stomachs or feces, but digestive erosion hampers species identification and underestimates fish weight (FW). To compensate, FW is often estimated from the least affected otoliths and scaled to other otoliths, which also introduces bias. The sulcus, an otolith surface feature, has a species‐specific shape of its ostium and caudal extents, which is within the otolith edge for some species. We explored whether the sulcus could improve species identification and estimation of prey size using a case study of four sciaenid species targeted by fisheries and bottlenose dolphins in North Carolina. Methods were assessed first on otoliths from a reference collection (n = 421) and applied to prey otoliths (n = 5,308) recovered from 120 stomachs of dead stranded dolphins. We demonstrated in reference‐collection otoliths that cauda to sulcus length (CL:SL) could discriminate between spotted seatrout (Cynoscion nebulosus) and weakfish (Cynoscion regalis) (classification accuracy = 0.98). This method confirmed for the first time predation of spotted seatrout by bottlenose dolphins in North Carolina. Using predictive models developed from reference‐collection otoliths, we provided evidence that digestion affects otolith length more than sulcus or cauda length, making the latter better predictors. Lastly, we explored scenarios of calculating total consumed biomass across degrees of digestion. A suggested approach was for the least digested otoliths to be scaled to other otoliths iteratively from within the same stomach, month, or season as samples allow. Using the otolith sulcus helped overcome challenges of species identification and fish size estimation, indicating their potential use in other diet studies.