Resolvin D1 improves survival in experimental sepsis through reducing bacterial load and preventing excessive activation of inflammatory response.

Sepsis is characterized as an uncontrolled inflammatory response. Spite et al. (Nature 461(7268):1287-1291, 2009) had demonstrated that resolvin D2, which is derived from docosahexaenoic acid (DHA), improves survival in cecal ligation and puncture (CLP)-initiated sepsis and enhances bacterial clearance without immune suppression. Resolvin D1, which is also derived from DHA and homologous with resolvin D2, is an endogenous anti-inflammatory and proresolving lipid molecule. We sought to investigate the effects of resolvin D1 on sepsis and to explore the mechanism of action. Six-to-eight-week-old male C57BL/6 mice were randomly divided into three groups: the sham group underwent the sham operation followed by tail vein injection of vehicle (0.1 % ethanol); the CLP group received vehicle (0.1 % ethanol) after CLP; the resolvin D1 group received resolvin D1 (100 ng) after CLP. Blood, peritoneal lavage fluid, and organs of mice were harvested 24 h after treatment for cytokine analysis, cell counts, bacterial cultures, histopathological studies, and apoptosis quantification. Compared with the vehicle control group, the survival rate and bacterial clearance of mice with sepsis induced by CLP were improved after resolvin D1 treatment, but the numbers of neutrophils in peritoneal lavage fluid, the inflammatory cytokines, the phosphorylation of the nuclear factor-κB (NF-κB) (P65) pathway, and the apoptosis rate of CD3(+) T lymphocytes of the thymus were suppressed. Resolvin D1 treatment improved survival in mice with sepsis induced by CLP, enhanced organism bacterial clearance, suppressed the increase of the numbers of neutrophils in peritoneal lavage fluid, reduced the release of inflammatory cytokines, and decreased the apoptosis rate of CD3(+) T lymphocytes of the thymus. These results suggest that resolvin D1 may attenuate the degree of inflammatory reaction in sepsis caused by CLP, without harming the host defense response.

Pre-treatment with Cobra venom factor alleviates acute lung injury induced by intestinal ischemia-reperfusion in rats.

BACKGROUND: Previous studies have shown that complement activation is required for intestinal ischemia-reperfusion (IIR)-induced tissue damage. Cobra venom factor (CVF), a structural and functional homolog to the activated form of C3 (the central component of the complement system), can cause exhaustive activation of the alternative pathway and deplete the complement components. AIM: This study aims to investigate the effect of CVF pretreatment on acute lung injury induced by IIR in rats. MATERIALS AND METHODS: Lung injury was induced by clamping superior mesenteric artery (SMA) for 60 min followed by 4 h of reperfusion. CVF was given via the tail vein 24 h before the operation. RESULTS: Histological results as well as lung edema determination and permeability assay showed the severe damages were induced in the lungs of rats in the IIR group, accompanying with the increases in the levels of pulmonary malondialdehyde (MDA), myeloperoxidase (MPO) activity, intercellular adhesion molecule-1 (ICAM-1), interleukin (IL)-8. Remarkably, CVF pretreatment significantly attenuated the morphological lung injury, lung edema and lung permeability, reduced the increase of the levels of MDA, MPO, ICAM-1 and IL-8 induced by IIR. In addition, the severe damage of intestinal and elevation of plasma diamine oxidase activity in the IIR rats were significantly alleviated by CVF pretreatment. CONCLUSIONS: CVF pretreatment could significantly reduce the acute lung injury induced by IIR. The mechanism might include, at least in part, the inhibition of oxidant generation, infiltration of neutrophils, ICAM-1 expression and IL-8 release. CVF might be an efficient reagent for preventing the IIR injuries in clinical condition.

Transillumination-assisted orotracheal intubation: a comparison of the Bonfils fibrescope and the lightwand (Trachlight).

BACKGROUND: Because the Bonfils fibrescope has a semi-rigid optical stylet and is similar in shape to a lightwand, we aimed to evaluate and compare the efficacy of transillumination-assisted orotracheal intubation with the Bonfils fibrescope and the Trachlight(TM) lightwand in patients with normal airways. METHODS: As a preliminary investigation to form a basis for later studies, therefore, we performed a randomized, single-blind study of 300 patients with normal airways to compare the efficiency of Trachlight and transillumination-assisted Bonfils orotracheal intubation in these patients. In both groups, orotracheal intubation was performed using a transillumination technique. The first attempt and overall success rates of tracheal intubation, the times required, and any untoward effects were recorded. RESULTS: Although the overall success rates were similar for Bonfils and Trachlight intubations (97.3% and 98.7%, respectively), tracheal intubation was successful on the first attempt in 87.3% of patients with the Bonfils fibrescope compared with 95.3% of patients with the Trachlight (P < 0.05). The mean intubation time for the first attempt was 15 ± 5 s with the Bonfils fibrescope and 9 ± 2 s with the Trachlight (P < 0.001). Patients intubated using the Bonfils fibrescope also experienced significantly more sore throat and hoarseness than those intubated using the Trachlight. CONCLUSIONS: For patients with normal airways, the Trachlight is superior for orotracheal intubation with respect to reliability, rapidity, and safety compared with the Bonfils fibrescope used with the transillumination technique.

Haplotypic variation and characteristics across the toll-like receptor 3 locus in chickens.

Toll-like receptor 3 (TLR3) has an important protective function against viral infection. The ability of an individual to respond properly to TLR ligands may be impaired by variants located in the TLR genes. By directly PCR sequencing four exons and their flanking sequence of chicken TLR3, a total of 50 nucleotide variants were identified from five breeds. Tibetan chickens and Silkies exhibited more abundant variation sites and rare alleles. Thirty haplotypes were reconstructed, with 31 variants whose minor allelic frequency was above 5% in five breeds, which revealed four divergent clades. Chicken TLR3 was partitioned into three haplotype blocks by the htSNPer program, and six tag SNPs could be used to distinguish these 30 haplotypes. Thirty variants were located in the coding sequence of chicken TLR3, and 16 of them were non-synonymous substitutions. It is predicted that p.Ser180Gly amino substitution could form an N-myristoylation site; the p.Lys240Thr amino substitution in chicken TLR3 could result in the loss of one protein kinase C phosphorylation site. These data provide a basic understanding of chicken TLR3 sequence variation and provide haplotypic markers for disease association studies.

Mapping quantitative trait loci affecting chicken body size traits via genome scanning.

Body size traits reflect the condition of body development, are always mentioned when a breed is described, and are also targets in breeding programmes. In chicken, there are several reports focused on body size traits, such as shank length, tibia length or bone traits. However, no study was carried out on chest width (CW), chest depth (CD), body slope length (BL) and head width (HW) traits. In this study, genome scans were conducted on an F(2) resource population (238 F(2) individuals from 15 full-sib families derived from an intercross of the White Plymouth Rock with the Silkies Fowl) to identify quantitative trait loci (QTL) associated with CW, CD, BL and HW from 7 to 12 weeks of age. In total, 21 significant or suggestive QTL were found that affected four body size traits. Four QTL reached 1% genome-wide significance level: at 297 cM on GGA3 (associated with CW at 9 weeks of age), between 155 and 184 cM on GGA1 (affecting BL traits at 9 and 10 weeks of age), at 22 cM on GGA2 (related with BL traits at 12 weeks of age) and at 36 cM on GGA1 (for HW trait at 8 weeks of age).

Structural analysis of a 4414-bp element in Drosophila melanogaster.

We cloned a 4414-bp element from a mutant of Drosophila melanogaster. Its insertion site was 18,929,626 bp. Analysis of the nucleotide and amino acid sequences demonstrated that the element is homologous to Pifo_I, first obtained from D. yabuka, which belongs to the gypsy/Ty3 subfamily. We also obtained a 3754-bp length element from a wild-type fly by PCR, with a pair of primers designed from the conserved region of the 4414-bp length element. The two elements included a pair of long terminal repeats and part of the GAG and ENV proteins, but the POL protein was completely lost. This element is found in the subgenus of D. melanogaster, but it is a degenerate type of Pifo_I and is not infective. Also, a 714-bp region structured in 5.0 tandem repeats of 143 bp each was found in the 5'UTR of the degenerate element; these could interact with transcription factor CF2. Phylogenetic analysis and alignment of amino acids indicated that the Pifo_I element was closer to the ZAM retrotransposon, which gave us some clues to their functional similarity. Based on these data, we propose that there is a relationship between the degenerate element and the mutant phenotype, which would provide a foundation for further research.

Expression and activity analysis reveal that heme oxygenase (decycling) 1 is associated with blue egg formation.

Biliverdin is responsible for the coloration of blue eggs and is secreted onto the eggshell by the shell gland. Previous studies confirmed that a significant difference exists in biliverdin content between blue eggs and brown eggs, although the reasons are still unknown. Because the pigment is derived from oxidative degradation of heme catalyzed by heme oxygenase (HO), this study compared heme oxygenase (decycling) 1 (HMOX1), the gene encoding HO expression and HO activity, in the shell glands of the Dongxiang blue-shelled chicken (n = 12) and the Dongxiang brown-shelled chicken (n = 12). Results showed that HMOX1 was highly expressed at the mRNA (1.58-fold; P < 0.05) and protein levels in blue-shelled chickens compared with brown-shelled chickens. At the functional level, blue-shelled chickens also showed 1.40-fold (P < 0.05) higher HO activity than brown-shelled chickens. To explore the reasons for the differential expression of HMOX1, an association study of 6 SNP capturing the majority of HMOX1 variants with the blue egg coloration was performed. Results showed no significant association between SNP and the blue egg coloration in HMOX1 (P > 0.05). Taken together, these results show that blue egg formation is associated with high expression of HMOX1 in the shell gland of Dongxiang blue-shelled chickens, and suggest that differential expression of HMOX1 in the 2 groups of chickens is most likely to arise from an alteration in the trans-acting factor.

Identification of quantitative trait loci for shank length and growth at different development stages in chicken.

Shank length affects chicken leg health and longer shanks are a source of leg problems in heavy-bodied chickens. Identification of quantitative trait loci (QTL) affecting shank length traits may be of value to genetic improvement of these traits in chickens. A genome scan was conducted on 238 F(2) chickens from a reciprocal cross between the Silky Fowl and the White Plymouth Rock breeds using 125 microsatellite markers to detect static and developmental QTL affecting weekly shank length and growth (from 1 to 12 weeks) in chickens. Static QTL affected shank length from birth to time t, while developmental QTL affected shank growth from time t-1 to time t. Seven static QTL on six chromosomes (GGA2, GGA3, GGA4, GGA7, GGA9 and GGA23) were detected at ages of 2, 3, 4, 5, 6, 7, 9 and 12 weeks, and six developmental QTL on five chromosomes (GGA1, GGA2, GGA4, GGA5 and GGA23) were detected for five shank growth periods, weeks 2-3, 4-5, 5-6, 10-11 and 11-12. A static QTL and a developmental QTL (SQSL1 and DQSL2) were identified at GGA2 (between ADL0190 and ADL0152). SQSL1 explained 2.87-5.30% of the phenotypic variation in shank length from 3 to 7 weeks. DQSL2 explained 2.70% of the phenotypic variance of shank growth between 2 and 3 weeks. Two static and two developmental QTL were involved chromosome 4 and chromosome 23. Two chromosomes (GGA7 and GGA9) had static QTL but no developmental QTL and another two chromosomes (GGA1 and GGA5) had developmental QTL but no static QTL. The results of this study show that shank length and shank growth at different developmental stages involve different QTL.

Minimum local analgesic concentration of ropivacaine for intra-operative caudal analgesia in pre-school and school age children.

We compared the minimum local analgesia concentration of ropivacaine for intra-operative caudal analgesia in pre-school and school age children. Fifty-one boys, undergoing hypospadius repair surgery, were stratified into pre-school or school age groups. After induction of anaesthesia, caudal block was performed with ropivacaine 1 ml.kg⁻¹ of the desired concentration. The first child in each group received ropivacaine 0.125%, and subsequent concentrations were determined by the analgesic response of the previous patient using Dixon's up-and-down method. Under general anaesthesia with 0.7 minimum alveolar concentration of sevoflurane, the minimum local analgesia concentration of ropivacaine for intra-operative caudal block was 34% greater in school age than in pre-school age boys (0.143% (95% CI 0.132-0.157%) vs 0.107% (95% CI 0.089-0.122%), respectively; p < 0.001). This study indicates that a higher concentration of ropivacaine is needed for school age than pre-school age children to provide intra-operative caudal analgesia when combined with general anaesthesia.

Localisation of the genomic sequence interval for the blue eggshell gene using an F2 resource population of Dongxiang chickens.

1. In order to identify the molecular interval containing the blue shell gene (O locus), linkage analysis was conducted with three microsatellite markers, (TTA)(n), (TG)(n) and (tg)(n), and a SNP in intron 1 of SLCO1C1 (solute carrier organic anion transporter family, member 1C1; A locus) to map the O locus in an F2 resource population of Dongxiang chickens. 2. Linkage analysis based on 98 F2 hens resulted in estimation of the best map order of the O locus with other linked markers as: (TTA)(n)-(TG)(n)-A-O-(tg)(n). 3. Based on these results, we inferred that the O locus was located between the A and (tg)(n) loci, that is, Chr1:67,296,991-69,140,571, which is the first genomic sequence interval to be established for the blue eggshell gene.

The influence of tube length, radius and chirality on the buckling behavior of single-walled carbon nanotubes filled with copper atoms.

The buckling behavior of single-walled carbon nanotubes completely filled with copper atoms under uniaxial compression is investigated using molecular dynamics simulations and compared with that modeled by continuum mechanics. The effects of geometrical characteristics, i.e. tube length, radius and chirality, on buckling deformations are explored separately. Results show that the behavior of encapsulated tubes is more complicated than that of empty ones due to the accommodation of the internal metal atoms. There are both similarities and differences between the results obtained by the molecular dynamics method and continuum mechanics. For a group of completely filled (10, 10) tubes with different length, the dependence of the critical strain on the tube length can be roughly divided into four different linear stages and is accompanied by a transition of the buckling mode from local to global. It is the competition between the evolution of the structure of metal atoms and the variation of the tube length that determines the critical strain. There exists a rather wide range of tube radii within which the critical strain has a weak dependence on tube radius, which differs from the observation for empty tubes. As compared with a zigzag tube of the same length and radius, an armchair tube has a lower critical strain but can be easily strengthened with the incorporation of internal metal atoms.

Mapping quantitative trait loci regulating chicken body composition traits.

Genome scans were conducted on an F(2) resource population derived from intercross of the White Plymouth Rock with the Silkies Fowl to detect QTL affecting chicken body composition traits. The population was genotyped with 129 microsatellite markers and phenotyped for 12 body composition traits on 238 F(2) individuals from 15 full-sib families. In total, 21 genome-wide QTL were found to be responsible for 11 traits, including two newly studied traits of proventriculus weight and shank girth. Three QTL were genome-wide significant: at 499 cm on GGA1 (explained 3.6% of phenotypic variance, P < 0.01) and 51 cm on GGA5 (explained 3.3% of phenotypic variance, P < 0.05) for the shank & claw weight and 502 cm on GGA1 (explained 1.4% of phenotypic variance, P < 0.05) for wing weight. The QTL on GGA1 seemed to have pleiotropic effects, also affecting gizzard weight at 490 cm, shank girth at 489 cm and intestine length at 481 cm. It is suggested that further efforts be made to understand the possible pleiotropic effects of the QTL on GGA1 and that on GGA5 for two shank-related traits.

Comparison of the total amount of eggshell pigments in Dongxiang brown-shelled eggs and Dongxiang blue-shelled eggs.

Based on the knowledge of the heme bio-synthetic and metabolic pathway and the structures of biliverdin and protoporphyrin, experiments were carried out to compare the difference between the total quality of eggshell pigments in blue-shelled eggs and brown-shelled eggs from the same population (Dongxiang, China) and to analyze the correlation between the quantity of protoporphyrin and biliverdin in the 2 kinds of eggshells. It was found that there was no significant difference between the total quantity of eggshell pigments in Dongxiang blue-shelled eggs and Dongxiang brown-shelled eggs (P = 0.9006), and a highly significant positive correlation between the quantity of protoporphyrin and biliverdin in blue eggshells (P < 0.01) and a significant positive correlation between the quantity of protoporphyrin and biliverdin in brown eggshells (P < 0.05). These results suggested that eggshell protoporphyrin and eggshell biliverdin probably derived from common precursor material.

Effect of selection for phagocytosis in dwarf chickens on immune and reproductive characters.

In current study, phagocytosis product (PP) of peripheral blood monocytes was detected among 920 dwarf chickens (460 per sex) at 20 wk of age, and based on discrepancies of PP, the flock was grouped (the highest group, the medium group, and the lowest group). Then serum hemagglutination inhibition antibody titers and subpopulations of T-lymphocytes of each group were examined after inoculations of avian influenza virus H5N2 inactivated vaccine (20 wk of age), avian influenza virus H9 inactivated vaccine (24 wk of age), and Newcastle disease virus-egg drop syndrome virus bigeminal inactivated vaccine (28 wk of age), respectively, to study the relationship between PP and immune response. To gain insight into effects of selection for PP on number of eggs, mean egg weight, fertilization rate, hatchability, and rate of healthy chicks, 9 (3 x 3) mating combinations were conducted. The results showed that (1) selection for higher PP in both sexes benefited to humoral immunity but not CD8(+) T-lymphocyte mediated immunity in dwarf chickens; (2) there were effects of selection for higher PP in hens on fertilization rate (P < 0.05), hatchability (P < 0.05), rate of healthy chicks (P < 0.05), and level of IgY antibody (P < 0.0001); however, hens' PP had no effects on number of eggs (P > or = 0.05) or egg weight (P > or = 0.05) and cocks' PP had no effect (P > or = 0.05) on any trait mentioned above. The results indicated that phagocytosis of peripheral blood monocytes might be an indicator of humoral immunity in dwarf chickens; furthermore, selection of hens with higher PP was not only beneficial to fertilization rate, but also benefited to hatchability and rate of healthy chicks in that the hens had stronger humoral immunity, which might contribute to maternal antibody in eggs.

Stronger in vitro phagocytosis by monocytes-macrophages is indicative of greater pathogen clearance and antibody levels in vivo.

Monocytes-macrophages are crucial players in specific and nonspecific immune responses to protect organisms from invasion of bacteria or viruses. In this study, monocytes in circulation from 2 lines of Silky and Starbro chickens with different disease resistance were separated and cultured in vitro. After identification with acridine orange (AO), Giemsa staining, and CD14 immunostaining, monocytes-macrophages were used for adherence and phagocytosis test. The overall percentages of adherence of Silky monocytes was 1.5 times greater than that of Starbro (P < 0.01), which were 26.85% +/- 8.24% and 18.34% +/- 8.15%, respectively (mean +/- SD). The monocytes-macrophages phagocytic index, phagocytic product, and percentage of phagocytosis in Silkies were greater than in Star-bros, respectively. The difference of phagocytic index was significant (P < 0.05), that is, 3.70 +/- 1.75 and 1.97 +/- 0.31, respectively (mean +/- SD). Then, 20 Silkies were divided into 2 groups according to phagocytic index: high phagocytic index (HPI) group and low phagocytic index (LPI) group, to study the relationship between phagocytic activity in vitro and pathogen clearance. After being challenged against Salmonella Pullorum C79-13, the Silky birds with HPI produced a 3-fold greater level of specific antibodies compared with those with LPI (P < 0.01), 50.21 +/- 6.67 and 16.85 +/- 4.52, respectively (mean +/- SD). In contrast to LPI birds, HPI birds shed less Salmonella Pullorum bacteria (P < 0.05), that is, 168.98 x 10(8) +/- 294.74 x 10(8) compared to 385.40 x 10(8) +/- 399.94 x 10(8) (mean +/- SD), and the shedding peak of Salmonella Pullorum in the test span appeared 4 d earlier. These results indicated that phagocytosis of monocytes-macrophages had strong effects on antibody titer and bacteria shedding postchallenge, which could be used to predict the disease resistance in animals.

Blood gas, hemoglobin, and growth of Tibetan chicken embryos incubated at high altitude.

Metabolism and hatchability are impaired when chicken eggs laid at sea level are incubated at high altitude. The Tibetan chicken is an excellent local poultry breed that inhabits altitudes of 2,900 m and has a hatchability of approximately 75% at that altitude. To understand how Tibetan chicken embryos develop successfully at high altitude, we compared blood gas, pH, hemoglobin concentrations and embryo mass for Tibetan chicken embryos (T) and for embryos from a dwarf breed (D) that normally is reared at sea level. The 2 breeds (T and D) and 2 incubation altitudes (2,900 m = high, H; and 100 m = low, L) were compared at 9, 12, 15, and 18 d of incubation. Embryo weights were lower for the high altitude groups (TH, DH) than for the low altitude groups at all stages of incubation. The embryo mass of TH appeared to increase more quickly than that of DH. Compared with DH, TH embryos had lower arterialized oxygen partial pressure on d 18, higher venous carbon dioxide partial pressure from d 12 to 18, and higher hemoglobin concentration and lower venous blood pH values on d 12 and 15. These findings indicate that the ability of the Tibetan chicken embryos to adapt to the high altitude may be due to the increase in hemoglobin concentration, which augments the blood oxygen-carrying capacity. In addition, the higher venous carbon dioxide partial pressure and lower venous blood pH promote unloading of oxygen from hemoglobin.

Study of the deposition process of eggshell pigments using an improved dissolution method.

An improved dissolution method called layer-by-layer dissolution was adopted to study the process of eggshell deposition, which is opposite to the process of eggshell dissolution. In the present study, blue and brown eggshells from 2 Chinese indigenous chicken breeds, Dongxing and Shouguang, respectively, were analyzed with layer-by-layer dissolution. The results showed that the deposition velocity of the eggshell pigments in the top (first) eggshell layer was the highest compared with other layers, which were biliverdin and protoporphyrin in blue eggshell or primarily protoporphyrin in brown eggshell. It was also revealed that the deposition processes of biliverdin and protoporphyrin were synchronous in the blue eggshell of the Dongxiang chicken in the present study.

A potential molecular marker for selection against abdominal fatness in chickens.

The peroxisome proliferators-activated receptor-gamma coactivator-1alpha (PGC-1alpha) was investigated as a candidate gene for growth and fatness traits in chicken because of its prominent role in muscle fiber specialization and adipogenesis. A single nucleotide polymorphism (SNP) from G to A at position 646 of the open reading frame of chicken PGC-1alpha gene causing an Asp216Asn amino acid substitution was identified. The frequencies of alleles and genotypes were significantly different among 6 chicken breeds (P < 0.01). The White Plymouth Rock had the highest frequency (0.67) of allele G, whereas the White Leghorn had the lowest (0.18). The associations of the SNP with the growth and fatness traits were evaluated in 332 F(2) birds from an experimental cross of White Plymouth Rock x Silkies. No association was found between the SNP and growth-related traits. However, abdominal fat weight at 12 wk of age for birds with genotype GG was 34.26 and 28.71% higher than those with genotypes AA and AG, respectively (P < 0.01), indicating that the Asp216Asn polymorphism of the PGC-1alpha gene could be used as a novel potential molecular marker for selection against abdominal fatness without interfering in regular breeding for growth rate of chickens.