A diagnosis model of parental care: How parents optimize their provisioning strategy in brood reduction?

Altricial birds often display biased preferences in providing parental care for their dependent offspring, especially during food shortages. During this process, such inflexible rules may result in provisioning errors. To demonstrate how parents optimize their provisioning strategies, we proposed a "diagnosis model" of parental care to posit that parents will undergo a diagnosis procedure to test whether selecting against some particular offspring based on phenotype is an optimal strategy. We tested this model in an asynchronous hatching bird, the Azure-winged Magpie Cyanopica cyanus, based on 10 years of data about demography and parental provisioning behaviors. Given their higher daily survival rates, core offspring (those hatched on the first day) merits an investment priority compared with their marginal brood mates (those hatched on later days). However, a marginal offspring also merited a priority if it displayed greater weight gain than the expected value at the early post-hatching days. Parents could detect such a marginal offspring via a diagnosis strategy, in which they provisioned the brood at the diagnosis stage by delivering food to every nestling that begged, then biased food toward high-value nestlings at the subsequent decision stage by making a negative response to the begging of low-value nestlings. In this provisioning strategy, the growth performance of a nestling became a more reliable indicator of its investment value than its hatching order or competitive ability. Our findings provide evidence for this "diagnosis model of parental care" wherein parents use a diagnosis method to optimize their provisioning strategy in brood reduction.

Nobiletin Inhibits Hypoxia-Induced Placental Damage via Modulating P53 Signaling Pathway.

In this study, we aimed to evaluate the effect of Nobiletin (NOB) on the placenta of Sprague-Dawley (SD) rats that had undergone reduced uterine perfusion pressure (RUPP) surgery and to evaluate the safety of NOB intervention during pregnancy. The results showed that NOB alleviated placental hypoxia, attenuated placental cell apoptosis, and inhibited placental damage in RUPP rats. No side effect of NOB intervention during pregnancy was observed. BeWo cell lines with P53 knockdown were then constructed using lentiviral transfection, and the P53 signaling pathway was found to be essential for NOB to reduce hypoxia-induced apoptosis of the BeWo cell lines. In summary, NOB attenuated hypoxia-induced placental damage by regulating the P53 signaling pathway, and those findings may contribute some insights into the role of NOB in placental development and the prevention of placental-related diseases.

Fitness consequences of divorce in the azure-winged magpie depends on the breeding experience of a new mate.

Sexual conflict in producing and raising offspring is a critical issue in evolutionary ecology research. Individual experience affects their breeding performance, as measured by such traits of provisioning of offspring and engagement in extra-pair copulations, and may cause an imbalance in sexual conflict. Thus, divorce is hypothesized to occur within aged social pairs, irrespective of current reproductive success. This concept was explored in the azure-winged magpie Cyanopica cyanus by investigating the divorce of a social pair and its relationship to their changes in breeding performance with prior experience. Females engaging in extra-pair copulation may intensify sexual conflicts and may be the main reason for divorce. Once divorced, females repairing with an inexperienced male realized higher reproductive success than that repairing with an experienced male; males repairing with an experienced female realized higher reproductive success than that repairing with an inexperienced female. This finding indicates that the fitness consequence of divorce depends on the breeding experience of new mates. Divorced females can obtain more extra-pair copulations, whereas divorced males cannot, when they repair with inexperienced breeders. Divorced females provisioned a brood at lower rates than inexperienced females whereas divorced males had no such difference. It appears that divorced females can obtain an advantage in sexual conflicts with inexperienced mates in future reproduction. Consequently, females are probably more active than males in divorcing their aged mates so as to select an inexperienced male as a new mate. Azure-winged magpies thus provide novel insights into the implications of sexual conflict in birds.

Trial marriage model-Female mate choice under male interference.

In sexually reproducing animals, the process of mate choice by females is often mixed with the process of male-male competition. Current models of female male choice focus mainly on how females identify the higher quality of males, but neglect the effect of male-male competition on the mate choice of females. Therefore, it remains controversial what is the relative importance of two processes in forming a social bond. We propose a new 'trial marriage' model for females' mate choice. The model assumes that females unconditionally accept any male they first encounter as their mating partner, and then conditionally switch mates to a new male of higher quality than their current partner when male-male competition occurs. This model was tested in the green weevil Hypomeces squamosus by exploring how females switched mates when males' mating interference was experimentally induced. The likelihood that females switched mates, as well as their conditional acceptance criteria of a new mate, was both raised with the intensity of males' mating interference that was manipulated in an enhanced encounter rate experiment, and in male introduction or stepwise removal experiments. These experimental findings confirm that a 'trial marriage' strategy occurs during females' mate choice. Compared with other strategies, it is more beneficial for females to choose a better mate without paying the costs of identifying males as suggested by the 'trial marriage' strategy. More importantly, using the current partner quality as the conditional acceptance threshold of new mates, females can choose better males in future encounters with potential mates. In the green weevils, males' preference for larger females and the higher possibility of the largest male winning an interference are mixed together when males' mating interference reaches a higher intensity. Therefore, the consequence of a male interference will determine which male could be chosen by a female. Under this condition, conditional acceptance of the winner becomes the most beneficial strategy of females in choosing their mates. We thus suggest that the 'trial marriage' strategy would be more efficient when males' mating interference becomes the determinant factor of females' mate choice.

Parental dependence on the nest's spatial cues in offspring recognition decreases with nestling growth in the azure-winged magpie.

In altricial birds, to address which cues are used by parents to recognize their offspring, and when they switch between cues during reproduction, it has not been well determined. In this study, we address this question in a Tibetan population of the azure-winged magpie Cyanopica cyanus, by examining the dependence of parents on a nest's spatial position in offspring recognition. During the egg and nestling phases, azure-winged magpie nests were translocated to new positions across various distances from their original site, and parental responses to the translocated nests were investigated. Our findings show that a nest's spatial position is not connected with the survival of its young, but might be used as a cue in parental offspring recognition. When nests are translocated to a new position within a certain distance, parents could recognize their nests and returned to resume their parenting behaviors. Parental dependence on the nest's spatial position in offspring recognition is higher during the egg phase than during the nestling phase, and it decreases with the growth of nestlings. After nestlings reach a certain age, the nest' s spatial position was no longer used by parents as the single cue for offspring recognition. These findings suggest that azure-winged magpies switch their cues in offspring recognition during the different stages of reproduction. After parent-offspring communication has been established, the offspring's phenotypic traits may become a more reliable cue than the nest's spatial position in offspring recognition.

Corticimicrobacter populi gen. nov., sp. nov., a member of the family Alcaligenaceae, isolated from symptomatic bark of Populus × euramericana canker.

One Gram-negative aerobic bacterial strain was isolated from the bark tissue of Populus × euramericana and investigated using a polyphasic approach including 16S rRNA gene sequencing and both phenotypic and chemotaxonomic assays. The 16S rRNA gene and housekeeping gene phylogenies suggest that the novel isolate is different from the other genera of the family Alcaligenaceae. The G+C content, major fatty acids, physiological and biochemical data supported the distinctiveness of the novel strain from reference species. The major fatty acids detected in the novel isolate were C16 : 1ω7c and/or C16 : 1ω6c, C16 : 0, C14 : 0 3OH and/or C16 : 1isoI and C18 : 1ω7c. The phospholipid profile of strain d3-2-2T contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminolipid, aminophospholipid and an unidentified lipid. The quinone of the novel isolate was Q-8. Therefore, based on the data, the strain constitutes a novel species of a novel genus within the family Alcaligenaceae, for which the name Corticimicrobacter populi gen. nov., sp. nov. is proposed. The type strain is 3d-2-2T (=CFCC 11891T=KCTC 52807T).

Whole-genome scanning for the heat-resistance-associated genes in the Droughtmaster breed (Bos taurus).

The Droughtmaster is a tropical breed of beef cattle developed in North Queensland that exhibits a combination of heat resistance and parasitic resistance from long-term artificial selection. Therefore, we used next-generation sequencing technology to screen the chromosomal regions and genes related to heat-resistance in cattle to provide data for improving cattle breeding. A total of 15,569,067 variants including 14,249,316 SNPs were obtained from two mixed pools by genome-wide resequencing. According to the results of the selective sweep analysis of the Droughtmaster pool compared to the nonheat resistant breeds pool, 81 candidate genes under selection in Droughtmaster were identified by combining Z HP and F ST analyses with a threshold standard of the top 1%, including SLC7A11, GYPC, and GYPC. In addition, 40 GO terms and 44 pathways were annotated from newly identified candidate genes. These signaling pathways were involved in environmental information processing, organismal systems, and metabolism. A majority of these genes have not been implicated in the previous studies of heat resistance. This study explored the genomic changes that result from long-term artificial selection, our findings help to explain the molecular mechanism of heat resistance in cattle.

Polymeric Ionic Liquid-Based Fluorescent Amphiphilic Block Copolymer Micelle for Selective and Sensitive Detection of p-Phenylenediamine.

A highly sensitive and selective detection of p-phenylenediamine (PPD) is achieved by a fluorescence sensor, which is constructed by encapsulating the hydrophobic fluorescent 1-pyrenecarboxaldehyde (Py-CHO) into a polymeric ionic liquid (PIL)-based amphiphilic block copolymer (BCP) micelle. The amine-aldehyde condensation reaction between PPD and Py-CHO leads to the fluorescence quenching of Py-CHO, giving rise to the basis for the quantitative detection of PPD. The core cavity of the micelle formed by the self-assembly of PIL provides an excellent hydrophobic environment for the accommodation of fluorescent Py-CHO, offering significant improved sensitivity and selectivity for PPD detection. The amount of PIL in fabricating the amphiphilic BCP micelle, the BCP-Py-CHO micelle concentration, and the detection pH condition are investigated to obtain the best performance of this sensor. The accurate detection of PPD is achieved in the range of 0.02-10 µmol L-1 under optimal conditions, and the detection limit is 0.007 µmol L-1 (3σ/ s). The developed sensor is successfully applied to the determination of PPD contents in hair dyes, spiked water, and urine samples.

Global DNA methylation was changed by a maternal high-lipid, high-energy diet during gestation and lactation in male adult mice liver.

An epigenetic mechanism has been suggested to explain the effects of the maternal diet on the development of disease in offspring. The present study aimed to observe the effects of a maternal high-lipid, high-energy (HLE) diet on the DNA methylation pattern of male offspring in mice. Female C57BL/6J mice were fed an HLE diet during gestation and lactation. The genomic DNA methylations at promoter sites of genes in the liver, mRNA and protein levels of selected genes related to lipid and glucose metabolism were measured by microarray, real-time PCR and Western blot. The results indicated that the percentage of methylated DNA in offspring from dams that were fed an HLE diet was significantly higher than that from dams that were fed a chow diet, and most of these genes were hypermethylated in promoter regions. The nuclear protein content and mRNA levels of hypermethylated genes, such as PPARγ and liver X receptor α (LXRα), were decreased significantly in offspring in the HLE group. The results suggested that the DNA methylation profile in adult offspring livers was changed by the maternal HLE diet during gestation and lactation.

Small interfering RNA survivin and GRIM-19 co-expression salmonella plasmid inhibited the growth of laryngeal cancer cells in vitro and in vivo.

OBJECTIVE: To investigate the inhibitory effect of plasmid-based survivin-specific short hairpin RNA and GRIM-19 on the growth of Hep-2 laryngeal cancer cells. METHODS: The plasmid expressing survivin-specific short hairpin RNA (shRNA) and GRIM-19 (p-siRNA survivin/GRIM-19) was prepared and transfected into Hep-2 cells with Lipofectamine 2000. The mRNA and protein expression of surviving and GRIM-19 were measured with RT-PCR and western blot assay, respectively. MTT assay was employed to detect the proliferation of Hep-2 cells, and flow cytometry and AO/EB assay were done to determine the apoptosis of Hep-2 cells. RESULTS: In the p-siRNA survivin/GRIM-19, the mRNA and protein expression of survivin was markedly reduced by 54.4% and 42.2%, and the reduction in protein expression of surviving was more obvious than that in the p-siRNA survivin group (37%) (P<0.05). The protein expression of GRIM-19 was markedly enhanced when compared with the control group (P<0.01). MTT assay revealed the proliferation of Hep-2 cells undergoing transfection with p-siRNA survivin/GRIM-19 was markedly inhibited, and the inhibition rate was as high as 79%, which was higher than that in the psi-survivin group (45%) and p-GRIM-19 group (35%). AO/EB assay and flow cytometry indicated that the apoptotic cells in the p-siRNA survivin/GRIM-19 group were dramatically increased as compared to the psi-survivin group and p-GRIM-19 group. CONCLUSION: The p-siRNA survivin/GRIM-19 has marked decrease in survivin expression and dramatic increase in GRIM-19 expression. Moreover, silencing of survivin and over-expression of GRIM-19 can significantly inhibit the growth and induce the apoptosis of Hep-2 in vitro and in vivo.

Adaptive responses by mouse fetus to a maternal HLE diet by downregulating SREBP1: a microarray- and bio-analytic-based study.

Maternal diet has long been recognized as a significant factor affecting offspring development and health, but the target genes affected by a maternal high-lipid diet are currently unknown. In this study, the gene expression profile of neonatal mouse liver was analyzed using gene chips to identify genes with significant up- or downregulated expression levels due to maternal high-fat diet during gestation. Real-time PCR and Western blotting were used to measure key genes selected using microarray. Serum lipid, glucose, and insulin levels in adult offspring from dams fed with chow or a high-lipid diet were measured using commercial kits. Results indicate that the expression of genes involved in cholesterol and fatty acid synthesis were significantly inhibited, while the expression of genes involved in glycolysis were significantly decreased by maternal high-lipid diet during gestation. SREBP1 might be the key gene regulating genes involved in fatty acid, glucose, and cholesterol metabolism in response to a maternal high-fat diet.

The effects of phytosterol supplementation on serum LDL-C levels and learning ability in mice fed a high-fat, high-energy diet from gestation onward.

A high-fat, high-energy (HFE) diet may be deleterious to the cardiovascular system and mental health. We previously reported that serum cholesterol levels and escape latency were significantly increased in mice by feeding them an HFE diet from gestation onward. In this study, we examined whether an HFE diet supplemented with phytosterols fed to pregnant C57BL/6j dams and their offspring would protect the HFE-diet-induced compromise of the offspring's learning capability. We measured serum cholesterol levels, brain N-methyl-D-aspartate receptor (NMDAR1) mRNA and protein expression and liver sterol 27-hydroxylase (Cyp27a1) mRNA expression, as well as a Morris water maze performance. The results showed that, compared to mice consuming the HFE diet alone, those also consuming phytosterols (the HFE + PS diet) significantly decreased mean serum low-density lipoprotein cholesterol levels and altered brain NMDAR1 mRNA and protein expression and liver Cyp27a1 mRNA expression. The Morris water maze experiments indicated that dietary phytosterol supplementation slightly decreased the escape latency (p = 0.07). Collectively, these observations suggest that consumption of phytosterols from early in life may help alleviate the detrimental effects of HFE diets in mice.

[Integration and "Overview" analysis of QTLs related to plant height in soybean].

Plant height, one of the most important factors for plant architectures, is closely related with some important traits such as plant density, lodging resistance and yields.. In the present study, 201 QTLs related to plant height of soybean from database and references were analyzed. The results revealed that 15 consensus QTLs located on chromosomes 6, 7, 11, 13, and 18 were identified after physical integrations and Meta analysis using the software BioMercator 2.1. Moreover, "Overview" method was used to optimize these QTLs based on statistic analysis. Some of the valid QTL regions were narrowed down to the 0.1 cM and mapped on certain chromosomes. Furthermore, the functions of all genes located in these confidence regions were predicted and 17 candidate genes that might be related to plant height were identified. These results would lay a foundation for fine mapping of QTLs related to plant height and marker-assisted breeding in soybean.

Plasmid-based Survivin shRNA and GRIM-19 carried by attenuated Salmonella suppresses tumor cell growth.

Persistent activation of Survivin and its overexpression contribute to the formation, progression and metastasis of several different tumor types. Therefore, Survivin is an ideal target for RNA interference mediated-growth inhibition. Blockade of Survivin using specific short hairpin RNAs (shRNA) can significantly reduce prostate tumor growth. RNA interference does not fully ablate target gene expression, owing to the idiosyncrasies associated with shRNAs and their targets. To enhance the therapeutic efficacy of Survivin-specific shRNA, we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 (GRIM-19). Then, the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium (S. typhimurium) to treat prostate cancer in vitro and in vivo. We found that the co-expressed Survivin-specific shRNA and GRIM-19 synergistically and more effectively inhibited prostate tumor proliferation and survival, when compared with treatment with either single agent alone in vitro and in vivo. This study has provided a novel cancer gene therapeutic approach for prostate cancer.

Effects of continuous haemofiltration on serum enzyme concentrations, endotoxemia, homeostasis and survival in dogs with severe heat stroke.

AIM: To examine the effectiveness of continuous haemofiltration as a treatment for severe heat stroke in dogs. METHODS: Dogs were randomly allocated to a control or continuous haemofiltration group (both n=8). Heat stroke was induced by placing anaesthetised dogs in a high temperature cabin simulator. Upon confirmation of heat stroke (rectal temperature>42 °C, mean arterial pressure (MAP) decrease>25 mmHg), dogs were removed from the chamber and continuous haemofiltration was initiated and continued for 3h for dogs in the continuous haemofiltration group. Dogs in the control group were observed at room temperature. RESULTS: Rectal temperature, haemodynamics, pH, blood gases and electrolyte concentrations rapidly returned to baseline in the continuous haemofiltration group, but not the control group. After 3h, rectal temperature was 36.68±0.51 °C in the continuous haemofiltration group and 39.83±1.10 °C in the control group (P<0.05). Continuous haemofiltration prevented endotoxin and all serum enzyme concentrations from increasing and caused malondialdehyde concentrations to decrease. After 3h, endotoxin concentrations were 0.14±0.02 EU ml(-1) in the continuous haemofiltration group and 0.23±0.05 EU ml(-1) in the control group (P=0.003), while malondialdehyde concentrations were 4.86±0.61 mmol l(-1) in the continuous haemofiltration group and 8.63±0.66 mmol l(-1) in the control group (P<0.001). Five dogs died in the control group within 3h, whereas no dogs died in the continuous haemofiltration group. CONCLUSIONS: Continuous haemofiltration rapidly reduced body temperature, normalised haemodynamics and electrolytes, improved serum enzyme concentrations and increased survival in dogs with heat stroke. Continuous haemofiltration may be an effective treatment for heat stroke.

Plasmid-based Stat3 siRNA delivered by hydroxyapatite nanoparticles suppresses mouse prostate tumour growth in vivo.

DNA vector-based Stat3-specific RNA interference (si-Stat3) blocks Stat3 signalling and inhibits prostate tumour growth. However, the antitumour activity depends on the efficient delivery of si-Stat3. The effects on the growth of mouse prostate cancer cells of si-Stat3 delivered by hydroxyapatite were determined in this study. RM-1 tumour blocks were transplanted into C57BL/6 mice. CaCl2-modified hydroxyapatite carrying si-Stat3 plasmids were injected into tumours, and tumour growth and histology were determined. The expression levels of Stat3, pTyr-Stat3, Bcl-2, Bax, Caspase3, VEGF and cyclin D1 were measured by western blot analysis. Amounts of apoptosis in cancer cells were analysed with immunohistochemistry and the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) assay. The results showed that hydroxyapatite-delivered si-Stat3 significantly suppressed tumour growth up to 74% (P < 0.01). Stat3 expression was dramatically downregulated in the tumours. The immunohistochemistry and TUNEL results showed that si-Stat3-induced apoptosis (up to 42%, P < 0.01). The Stat3 downstream genes Bcl-2, VEGF and cyclin D1 were also strongly downregulated in the tumour tissues that also displayed significant increases in Bax expression and Caspase3 activity. These results suggest that hydroxyapatite can be used for the in vivo delivery of plasmid-based siRNAs into tumours.

Weight-reducing effect of Acer truncatum Bunge may be related to the inhibition of fatty acid synthase.

In order to study the anti-obesity ability and inhibition towards fatty acid synthase (FAS) of the extract, a 70% ethanol extract of Acer truncatum Bunge (AT) leaves was further extracted with ethyl acetate. FAS is a very significant lipogenic enzyme, participating in energy metabolism in vivo; it has also been observed that FAS inhibitors might be potent anti-obesity agents. Experimental results on animals showed that the extract significantly reduced food intake and adipose, and effectively controlled weight evolution. Lipogenesis inhibition might be regarded as one of the reasons for the weight control and adipose reduction by AT. The extract was further isolated using a series of column chromatography that yielded 10 known compounds. 1,2,3,4,6-Penta-O-galloyl-ß-D-glucose was found to be one of the major active constituents in the extract of AT.

Knockdown of stat3 expression by RNAi inhibits in vitro growth of human ovarian cancer.

BACKGROUND: The aim of the study was to investigate the suppressive effects of pSilencer2.1-U6-siRNA-stat3 recombinant plasmids on the growth of ovarian cancer in vitro. MATERIAL AND METHODS.: Three pairs of DNA template (stat3-1, stat3-2, stat3-3) specific for different target sites on stat3 mRNA were synthesized to reconstruct pSilencer2.1-U6-siRNA-stat3s, which were transfected into SKOV3 cells. The expressions of STAT3, BcL-2, cyclin D1 and C-myc in these cells were detected by Western blot and Northern blot. The cell cycle and the growth were determined by flow cytometry (FCM) and MTT assay, respectively. Cell apoptosis was determined by TUNEL staining. RESULTS: Of the three siRNAs, only siRNA targeting stat3-3 markedly suppressed the protein expression of stat3 in SKOV3 cells; MTT assay and FCM showed that transfection of stat3-3 siRNA could significantly suppress the growth of SKOV3 cells and arrest the cell cycle in vitro. TUNEL staining also showed massive apoptosis in SKOV3 cells transfected with stat3-3 siRNA. CONCLUSIONS: pSilencer2.1-U6-siRNA-stat3-3 can significantly inhibit the STAT3 expression in human ovarian cancer cells resulting in the inhibition of the cancer growth and the increase of apoptosis of cancer cells.

[Suppressing the growth of Hep-2 human laryngeal cancer cells by silencing survivin gene in vitro and in vivo].

OBJECTIVE: To study the effect of silencing survivin on the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo. METHODS: Hep-2 cells were transfected with pGCsilencer-siRNA-survivin (psi-survivin)by Lipofectamine 2000. The mRNA and protein expressions of survivin were detected by semi-quantitative RT-PCR and Western blot, respectively. Cell proliferation activity was measured by MTT assay. Apoptosis was assessed by flow cytometry. The implanted tumors were formed from injected Hep-2 cells in nude mice. After the tumor formation, psi-survivin was injected into peritumor tissues. The growth of tumor were observed. The tumor volume was calculated and the tumor growth curve was plotted. The expression of survivin in tumor tissues was detected by Western blot. The tumor cell apoptosis was observed by Tunel staining. RESULTS: The sequence-specific siRNA of survivin inhibited the expressions of survivin mRNA and protein. The inhibition rates of survivin mRNA and protein expression were 54.4% and 37.0% respectively. Also the growth of Hep-2 cells was inhibited significantly, with a decrease by 71.7%. By the day 32 of tumor growth, the mean tumor volumes were (1443.9 ± 230.5) mm(3) (x(-) ± s) in saline control group, (1348.5 ± 198.4) mm(3) in plasmid-negative control group, and (624.6 ± 188.4) mm(3) in psi-survivin group, respectively (t = -5.917, P < 0.01). In the implanted tumors injected with psi-survivin, survivin protein expression was down-regulated significantly, with a inhibition rate of 41.8%. Tunel staining showed the apoptosis occurred in the implanted tumors. CONCLUSION: Silencing survivin could significantly inhibit the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo.