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11.
Innovative approaches to genome editing in avian species   总被引:1,自引:0,他引:1  
The tools available for genome engineering have significantly improved over the last 5 years, allowing scientist to make precise edits to the genome. Along with the development of these new genome editing tools has come advancements in technologies used to deliver them. In mammals genome engineering tools are typically delivered into in vitro fertilized single cell embryos which are subsequently cultured and then implanted into a recipient animal.In avian species this is not possible, so other methods have been developed for genome engineering in birds. The most common involves in vitro culturing of primordial germ cells(PGCs), which are cells that migrate through the embryonic circulatory system to the developing gonad and colonize the gonad, eventually differentiating into the gonadocytes which produce either sperm or ova. While in culture the PGCs can be modified to carry novel transgenes or gene edits, the population can be screened and enriched, and then transferred into a recipient embryo. The largest drawback of PGC culture is that culture methods do not transfer well across avian species, thus there are reliable culture methods for only a few species including the chicken. Two newer technologies that appear to be more easily adapted in a wider range of avian species are direct injection and sperm transfection assisted gene editing(STAGE).The direct injection method involves injecting genome engineering tools into the circulatory system of the developing embryo just prior to the developmental time point when the PGCs are migrating to the gonads. The genome engineering tools are complexed with transfection reagents, allowing for in vivo transfection of the PGCs. STAGE utilizes sperm transfection to deliver genome engineering tools directly to the newly fertilized embryo. Preliminary evidence indicates that both methodologies have the potential to be adapted for use in birds species other than the chicken, however further work is needed in this area.  相似文献   
12.
Birth in all higher vertebrates is at the center of the critical window of development in which newborns transition from dependence on innate immunity to dependence on their own adaptive immunity, with passive maternal immunity bridging this transition. Therefore we have studied immunological development through fetal and early neonatal life. In swine, B cells appear earlier in fetal development than T cells. B cell development begins in the yolk sac at the 20th day of gestation (DG20), progresses to fetal liver at DG30 and after DG45 continues in bone marrow. The first wave of developing T cells is gammadelta cells expressing a monomorphic Vdelta rearrangement. Thereafter, alphabeta T cells predominate and at birth, at least 19 TRBV subgroups are expressed, 17 of which appear highly homologous with those in humans. In contrast to the T cell repertoire and unlike humans and mice, the porcine pre-immune VH (IGHV-D-J) repertoire is highly restricted, depending primarily on CDR3 for diversity. The V-KAPPA (IGKV-J) repertoire and apparently also the V-LAMBDA (IGLV-J) repertoire, are also restricted. Diversification of the pre-immune B cell repertoire of swine and the ability to respond to both T-dependent and T-independent antigen depends on colonization of the gut after birth in which colonizing bacteria stimulate with Toll-like receptor ligands, especially bacterial DNA. This may explain the link between repertoire diversification and the anatomical location of primary lymphoid tissue like the ileal Peyers patches. Improper development of adaptive immunity can be caused by infectious agents like the porcine reproductive and respiratory syndrome virus that causes immune dysregulation resulting in immunological injury and autoimmunity.  相似文献   
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Pathogenic infections increase morbidity and reduce performance in livestock, and thus understanding the comprehensive physiological changes associated with infections can benefit production sustainability. In this study, we sought to investigate such physiological responses to an acute immune challenge in lambs. Polypay wethers received single IV injections of 1.5 µg/kg lipopolysaccharide endotoxin (LPS-injected; n = 6) or saline (controls; n = 6). Corneal temperatures (via infrared thermography), rectal temperatures, blood, plasma, and saliva were assessed every 2 hr for 10 hr after injections. Blood was also assessed at 24 hr. LPS-injected lambs exhibited elevated (P < 0.05) corneal and rectal temperatures that peaked at 4 hr but were still slightly greater (P < 0.05) than controls at 10 hr. Circulating total white blood cells, monocytes, and granulocytes were reduced (P < 0.05) in LPS-injected lambs within the first 4 hr but were subsequently greater (P < 0.05) than in controls. Lymphocytes were reduced (P < 0.05) in LPS-injected lambs over the first 8 hr and did not differ from controls thereafter. Red blood cells, hematocrit, and hemoglobin were increased (P < 0.05) in LPS-injected lambs over the first 6 hr, indicating mild dehydration. Blood glucose briefly increased (P < 0.05) in LPS-injected lambs at 2 hr but was less (P < 0.05) than in controls thereafter. Blood lactate was greater (P < 0.05) in LPS-injected lambs between 6 and 10 hr after injections, which together with reduced (P < 0.05) CO2 partial pressure indicated a metabolic shift toward glycolysis. LPS-injected lambs exhibited a transient increase (P < 0.05) in plasma TNFα at 2 and 4 hr only and sustained increases (P < 0.05) in CXCL9 and CXCL10 beginning at 6 and 4 hr, respectively. They also exhibited a mild, paradoxical increase (P < 0.05) in the anti-inflammatory sFRP3. Salivary TNFα was increased (P < 0.05) in LPS-injected lambs at 2 hr only. Regression analyses indicated that rectal temperatures were a generally poor predictor of the other inflammatory components in this study, with the exception of circulating leukocyte populations. Likewise, correlations among the 10 cytokines measured in this study were generally weak, with notable exceptions between CXCL9 and CXCL10 and between IL-21 and IFNγ. These findings demonstrate that physiological changes to even short-lived immune challenges are dynamic in nature and persist beyond the time frame of febrile responses and other common assessments.  相似文献   
15.
Intrauterine stress impairs growth and metabolism in the fetus and offspring. We recently found that sustained maternofetal inflammation resulted in intrauterine growth-restricted (MI-IUGR) fetuses with asymmetric body composition, impaired muscle glucose metabolism, and β-cell dysfunction near term. These fetuses also exhibited heightened inflammatory tone, which we postulated was a fetal programming mechanism for the IUGR phenotype. Thus, the objective of this study was to determine whether poor growth and metabolism persisted in MI-IUGR lambs after birth. Polypay ewes received serial lipopolysaccharide or saline injections in the first 2 wk of the third trimester of pregnancy to produce MI-IUGR (n = 13) and control (n = 12) lambs, respectively. Lambs were catheterized at 25 d of age. β-Cell function was assessed at 29 d, hindlimb glucose metabolism at 30 d, and daily blood parameters from day 26 to 31. Glucose metabolism was also assessed in flexor digitorum superficialis (FDS) muscle isolated at necropsy on day 31. Asymmetric body composition persisted in MI-IUGR neonates, as these lambs were lighter (P < 0.05) than controls at birth and 31 d, but body and cannon bone lengths did not differ at either age. FDS muscles from MI-IUGR lambs were smaller (P < 0.05) and exhibited reduced (P < 0.05) glucose oxidation and Akt phosphorylation but similar glucose uptake compared with controls when incubated in basal or insulin-spiked media. Similarly, hindlimb glucose oxidation was reduced (P < 0.05) in MI-IUGR lambs under basal and hyperinsulinemic conditions, but hindlimb glucose utilization did not differ from controls. Circulating urea nitrogen and cholesterol were reduced (P < 0.05), and triglycerides, high-density lipoprotein cholesterol, and glucose-to-insulin ratios were increased (P < 0.05) in MI-IUGR lambs. Glucose and insulin concentrations did not differ between groups during basal or hyperglycemic conditions. Although circulating monocyte and granulocyte concentrations were greater (P < 0.05) in MI-IUGR lambs, plasma tumor necrosis factor α (TNFα) was reduced (P < 0.05). FDS muscle contained greater (P < 0.05) TNF receptor 1 (TNFR1) and IκBα protein content. These findings indicate that maternofetal inflammation in late pregnancy results in fetal programming that impairs growth capacity, muscle glucose oxidation, and lipid homeostasis in offspring. Inflammatory indicators measured in this study appear to reflect heightened cytokine sensitivity in muscle and compensatory systemic responses to it.  相似文献   
16.
  1. The ocean's remoteness, ecological complexities, lengthy ecosystem processes, and vulnerability to multiple and cumulative anthropogenic threats make marine conservation communication particularly difficult. Both scientists and journalists face unique challenges in explaining the science of these often out-of-sight ecosystems. Given the inadequacies of marine news, improvement appeared necessary. However, the experiences and views of journalists and scientists have hardly been examined within marine news contexts. Thus, this study sought the perspectives of these two professional groups to provide a discussion on ways to improve news coverage of complex ocean issues through enhanced journalist–scientist working relations.
  2. Both journalists and scientists rated the quantity, breadth and quality of marine news as average and were receptive to alternative ways for reporting ecosystem complexities. Although some frustrations remain, both valued impactful news stories resulting from their working relations and preferred direct contact with each other over indirect methods such as press releases and science news platforms. Both groups generally agreed on what to include in marine news, but scientists favoured a collaborative approach to news content decision-making more strongly than journalists.
  3. Journalists' and scientists' commonly shared views and goals concerning marine news identified in this study could serve as a common ground for uniting the two professions. Institutional policies that permit one-on-one journalist–scientist interactions could lead to mutual understandings about the contexts of their relationship challenges. More trustful and mutually beneficial relationships, in turn, could be a basis for a more collaborative news generation process. Compiling and making marine visuals readily accessible to journalists; training programmes that enhance journalists' and scientists' understanding of the influence of media message framing on conservation actions; media appreciation of marine ecosystem complexities' newsworthiness; and the notion of media's social responsibility in reporting marine conservation issues could contribute to more impactful coverage.
  相似文献   
17.
Background: Hematologic and biochemical reference intervals depend on many factors, including age. A review of the literature highlights the lack of reference intervals for 6-wk-old specific pathogen free (SPF) Hampshire-Yorkshire crossbred pigs. For translational research, 6-wk-old pigs represent an important animal model for both human juvenile colitis and diabetes mellitus type 2 given the similarities between the porcine and human gastrointestinal maturation process. The aim of this study was to determine reference intervals for hematological and biochemical parameters in healthy 6-wk-old crossbred pigs. Blood samples were collected from 66 clinically healthy Hampshire-Yorkshire pigs. The pigs were 6 wks old, represented both sexes, and were housed in a SPF facility. Automated hematological and biochemical analysis were performed using an ADVIA 120 Hematology System and a Cobas 6000 C501 Clinical Chemistry Analyzer. Results: Reference intervals were calculated using both parametric and nonparametric methods. The mean, median, minimum, and maximum values were calculated. Conclusion: As pigs are used more frequently as medical models of human disease, having reference intervals for commonly measured hematological and biochemical parameters in 6-wk-old pigs will be useful. The reference intervals calculated in this study will aid in the diagnosis and monitoring of both naturally occurring and experimentally induced disease. In comparison to published reference intervals for older non SPF pigs, notable differences in leukocyte populations, and in levels of sodium, potassium, glucose, protein, and alkaline phosphatase were observed.  相似文献   
18.

Objective

To compare postanesthetic xylazine and dexmedetomidine on recovery characteristics from sevoflurane anesthesia in horses.

Study design

Randomized, crossover study.

Animals

Six geldings, mean ± standard deviation (SD) (range), 17 ± 4 (11–24) years and 527 ± 80 (420–660) kg.

Methods

Horses were anesthetized with sevoflurane for 60 minutes under standardized conditions for a regional limb perfusion study. In recovery, horses were administered either xylazine (200 μg kg?1) or dexmedetomidine (0.875 μg kg?1) intravenously. Recoveries were unassisted and were video-recorded for later evaluation of recovery events and quality by two individuals unaware of treatment allocation. Recovery quality was assessed using a 100 mm visual analog scale (VAS) (0 = poor recovery, 100 = excellent recovery), the Edinburgh Scoring System (ESS) (0–100; 100 = excellent recovery) and the mean attempt interval (MAI) (longer = better). Data are mean ± SD.

Results

All recovery quality assessments (xylazine and dexmedetomidine, respectively: VAS: 71 ± 21 mm, 84 ± 13 mm; ESS: 65 ± 22, 67 ± 30; MAI: 52 ± 24 minutes, 60 ± 32 minutes) and events (first limb movement: 37 ± 8 minutes, 42 ± 10 minutes; first attempt to lift head: 44 ± 12 minutes, 48 ± 9 minutes; first attempt to sternal posture: 57 ± 28 minutes, 50 ± 7 minutes; number of head bangs: 2.0 ± 3.0, 0.5 ± 0.5; time to first attempt to stand: 72 ± 6 minutes, 78 ± 13 minutes; time to standing: 79 ± 14 minutes, 84 ± 13 minutes) did not differ significantly between treatments (p > 0.05).

Conclusions and clinical relevance

Recovery characteristics did not differ significantly between postanesthetic xylazine and dexmedetomidine following 1 hour of sevoflurane anesthesia in horses in this study. Further evaluations in more horses and in younger horses are required to confirm these results.  相似文献   
19.
20.
There are few formal studies on the contribution of botanical gardens as urban green spaces, particularly within developing countries. Therefore, this paper reports on an assessment of the use and appreciation of botanical gardens as urban green spaces in South Africa. Users and staff were surveyed in six national botanical gardens. The gardens provided numerous benefits in terms of conservation, education and recreation. However, the people using the gardens were not demographically representative of the general population of the surrounding city or town. Generally, most of the visitors were middle- to old-aged, well-educated professionals with medium to high incomes. Most were white and English was their home language. There was an even gender representation. Most visited only a few times per year. The majority of users visited the gardens for recreation and psychological reasons rather than educational ones. However, the staff of each garden placed emphasis on education in the gardens and amongst surrounding schools. Most visitors appreciated the conservation dimensions of botanical gardens, and felt that there was insufficient public green space in their town or city. Understanding how people perceive and use the botanical gardens of South Africa is important to inform future research and strategies regarding the conservation of urban green space within a developing country.  相似文献   
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