Effects of packaging methods on the quality of heavy metals-free preserved duck eggs during storage.

Preserved eggs without adding heavy metals in the pickling solution (heavy metals-free preserved eggs) have been developed, but it was found that the undesirable phenomenon such as dry shrinkage and fading occurred when they were not packaged and stored at room temperature. In this study, the effects of 5 packaging methods on the quality of heavy metals-free preserved eggs during storage were systematically studied. These methods included storage at room temperature and 4°C without packaging, wrapping with plastic bags, paraffin coating, and vacuum package. Through adopting these 5 packaging methods, the results showed that the moisture content and pH of the albumen decreased continuously, the mass loss rate increased continuously, the content of total volatile basic nitrogen increased firstly and then decreased, and the albumen hardness increased continuously. No microorganisms were detected in all samples with the 5 packaging methods during storage. Among them, the uncoated preserved eggs suffered the most serious moisture loss and mass loss, and the pH dropped at the fastest rate, followed by the preserved eggs wrapped in plastic bags. Preserved eggs stored at low temperature tended to turn yellow during storage, and the albumen showed higher hardness. The packaging method of paraffin coating performed the best in preventing the moisture loss of the albumen and the weight loss, which only decreased by 0.34 and 1.24%, respectively, after 3 mo. The best springiness, the darkest color, and the highest sensory score were found in the vacuum-packed preserved eggs after 3 mo of storage. It was concluded that paraffin coating and vacuum packing had better effect, while plastic bag packing showed the worst preservation performance for heavy metals-free preserved eggs.

Changes in physicochemical properties, gel structure and in vitro digestion of marinated egg white gel during braising.

In this paper, changes in physicochemical properties, gel structure and in vitro digestion of marinated egg with spice or tea during braising were investigated. Results indicated that the moisture content and surface hydrophobicity of marinated egg white showed an overall decreased trend. The springiness of marinated egg white showed an increased trend, and the hardness in the late stage showed an increased trend. Microstructure showed that compact gel structures formed many holes during the braising. Intermolecular forces showed that ionic bonds and disulfide bonds played a dominant role in the marinated egg white. Secondary structure showed that the ß-turn showed a decreased trend, contrary to that of random coils and α-helices. Appropriate braising increased the digestibility of marinated egg white, but excessively long-time braising could reduce it. Both spice and tea braising could improve the gel strength of protein, and the tea braising was also slightly better than spice braising.

Effects of temperature on quality of preserved eggs during storage.

The effects of storage temperature (4°C, 25°C, and 35°C) on sensory quality, physicochemical properties, texture, molecular forces, flavor, and microbial indexes of preserved eggs were studied. The results showed that the sensory quality, weight loss rate, pH, and color of preserved eggs were significantly different at different storage temperatures (P < 0.05). Compared with high temperature and normal temperature storage, low temperature storage reduced weight loss rate by 55.15 and 64.1%, respectively, improved the sensory score (P < 0.05), inhibited the reduction of pH and the increase of total volatile base nitrogen (P < 0.05), and decreased the change of color (P < 0.05). During storage, there was no difference in the springiness of preserved egg white stored at different temperatures (P > 0.05). Hardness and chewiness at 3 different temperatures increased first and then decreased, and low temperature significantly inhibited the progress of these changes to a certain extent (P < 0.05). The content of ionic bond in egg white first decreased and then increased, and content of disulfide bond increased first and then decreased. Content of ionic bond in yolk decreased all the time, and high temperature could promote this change. Whatever the temperature was, the content of free amino acids in preserved egg white and yolk increased first and then decreased, and the total content of amino acids stored at different temperatures was significantly different (P < 0.05). The content of free fatty acids in yolk decreased. At the end of storage, no microorganisms were detected in 3 temperatures during the storage period of 84 D. The results showed that low temperature storage is more conducive for preservation of preserved eggs.

Thrombocytopenia in pregnancy with different diagnoses: Differential clinical features, treatments, and outcomes.

To investigate the clinical features and perinatal treatment of thrombocytopenia induced by different causes during pregnancy.Clinical data from 195 pregnant women with thrombocytopenia attending 2 tertiary hospitals from January 2014 to October 2016 were retrospectively studied. The obtained data were analyzed with SPSS 19.0 software.There were 117 (60.0%), 55 (28.2%), and 23 cases (11.8%) of pregnancy-associated thrombocytopenia (PAT), idiopathic thrombocytopenia (ITP), and hypertensive disorder in pregnancy (PIH), respectively. The percentage of nulliparous women, gestational age at delivery, date of diagnosis of thrombocytopenia, and delivery mode significantly differed between the patients in these 3 groups (P < .05). Patients with PIH had a higher percentage of premature delivery and of lower birth weight infants than patients in the other 2 groups. The 3 groups had similar incidences of postpartum hemorrhage, rates of stillbirth, and neonatal Apgar scores at 5 minutes. PAT and PIH patients had different platelet counts after delivery compared with at diagnosis, whereas the platelet counts of the ITP patients were similar at diagnosis and after delivery. ITP patients in the nontreatment group and the treatment group had significantly different platelet counts (P < .05), and in the treatment group, the maternal platelet count did not differ for treatment with intravenous immunoglobulin (IVIg) versus corticosteroids.The causes of thrombocytopenia in pregnancy are diverse, and the clinical features vary widely. Timely analysis is needed to determine the primary cause of thrombocytopenia, and appropriate therapy should then be selected to effectively improve the prognosis of pregnancies.

Cell line donor genotype and its influence on experimental phenotype: Toll-like receptor SNPs and potential variability in innate immunity.

Cell lines are used to model a disease and provide valuable information regarding phenotype, mechanism, and response to novel therapies. Derived from individuals of diverse genetic backgrounds, the cell's genetic complement predicts the phenotype, and although some lines have been sequenced, little emphasis has been placed on genotyping. Toll-like receptors (TLRs) are essential in initiating the inflammatory cascade in response to infection. TLR single nucleotide polymorphism (SNP) alleles may predict an altered innate immune response: a SNP can affect TLR-dependent pathways and may alter experimental results. Thus, genotype variation may have far-reaching implications when using cell lines to model phenotypes. We recommend that cell lines be genotyped and cataloged in a fashion similar to that used for bacteria, with cumulative information being archived in an accessible central database to facilitate the understanding of SNP cell phenotypes reported in the literature.

Induction of chemokines and prostaglandin synthesis pathways in luteinized human granulosa cells: potential role of luteotropin withdrawal and prostaglandin F2α in regression of the human corpus luteum.

Our objective was to determine the effects of prostaglandin F2α (PGF2α) and withdrawal of luteotropic stimulants (forskolin or hCG) on expression of chemokines and prostaglandin-endoperoxide synthase 2 (PTGS2) in luteinized human granulosa cells. Human granulosa cells were collected from 12 women undergoing oocyte retrieval and were luteinized in vitro with forskolin or hCG. In first experiment, granulosa-lutein cells were treated with PGF2α, the primary luteolytic hormone in most species. In second experiment, granulosa cells that had been luteinized for 8 d had luteotropins withdrawn for 1, 2, or 3 d. Treatment with PGF2α induced mRNA for chemokine (c-x-c motif) ligand 2 (CXCL2) and CXC ligand 8 (CXCL8; also known as interleukin-8) in granulosa cells luteinized for 8 d but not in cells that were only luteinized for 2 d. Similarly, luteinization of human granulosa cells for 8 d with forskolin or hCG followed by withdrawal of luteotropic stimulants, not only decreased P4 production, but also increased mRNA concentrations for CXCL8, CXCL-2 (after forskolin withdrawal), and PTGS2. These results provide evidence for two key steps in differentiation of luteolytic capability in human granulosa cells. During 8 d of luteinization, granulosa cells acquire the ability to respond to luteolytic factors, such as PGF2α, with induction of genes involved in immune function and PG synthesis. Finally, a decline in luteotropic stimuli triggers similar pathways leading to induction of PTGS2 and possibly intraluteal PGF2α production, chemokine expression, leukocyte infiltration and activation, and ultimately luteal regression.

Oxytocin expression and function in the posterior retina: a novel signaling pathway.

PURPOSE: Oxytocin (OXT) is recognized as an ubiquitously acting nonapeptide hormone that is involved in processes ranging from parturition to neural development. Its effects are mediated by cell signaling that occurs as a result of oxytocin receptor (OXTR) activation. We sought to determine whether the OXT-OXTR signaling pathway is also expressed within the retina. METHODS: Immunohistochemistry using cell-specific markers was used to localize OXT within the rhesus retina. Reverse transcriptase PCR and immunohistochemistry were used to assess the expression of OXTR in both human and rhesus retina. Single-cell RT-PCR and Western blot analyses were used to determine the expression of OXTR in cultured human fetal RPE (hfRPE) cells. Human fetal RPE cells loaded with FURA-2 AM were studied by ratiometric Ca(2+) imaging to assess transient mobilization of intracellular Ca(2+) ([Ca(2+)]i). RESULTS: Oxytocin was expressed in the cone photoreceptor extracellular matrix of the rhesus retina. Oxytocin mRNA and protein were expressed in the human and rhesus RPE. Oxytocin mRNA and protein expression were observed in cultured hfRPE cells, and exposure of these cells to 100 nM OXT induced a transient 79 ± 1.5 nM increase of [Ca(2+)]i. CONCLUSIONS: Oxytocin and OXTR are present in the posterior retina, and OXT induces an increase in hfRPE [Ca(2+)]i. These results suggest that the OXT-OXTR signaling pathway is active in the retina. We propose that OXT activation of the OXTR occurs in the posterior retina and that this may serve as a paracrine signaling pathway that contributes to communication between the cone photoreceptor and the RPE.

Patterns of gene expression in the bovine corpus luteum following repeated intrauterine infusions of low doses of prostaglandin F2alpha.

Natural luteolysis involves multiple pulses of prostaglandin F2alpha (PGF) released by the nonpregnant uterus. This study investigated expression of 18 genes from five distinct pathways, following multiple low-dose pulses of PGF. Cows on Day 9 of the estrous cycle received four intrauterine infusions of 0.25 ml of phosphate-buffered saline (PBS) or PGF (0.5 mg of PGF in 0.25 ml of PBS) at 6-h intervals. A luteal biopsy sample was collected 30 min after each PBS or PGF infusion. There were four treatment groups: Control (n = 5; 4 PBS infusions), 4XPGF (4 PGF infusions; n = 5), 2XPGF-non-regressed (2 PGF infusions; n = 5; PGF-PBS-PGF-PBS; no regression after treatments), and 2XPGF-regressed (PGF-PBS-PGF-PBS; regression after treatments; n = 5). As expected, the first PGF pulse increased mRNA for the immediate early genes JUN, FOS, NR4A1, and EGR1 but unexpectedly also increased mRNA for steroidogenic (STAR) and angiogenic (VEGFA) pathways. The second PGF pulse induced immediate early genes and genes related to immune system activation (IL1B, FAS, FASLG, IL8). However, mRNA for VEGFA and STAR were decreased by the second PGF infusion. After the third and fourth PGF pulses, a distinctly luteolytic pattern of gene expression was evident, with inhibition of steroidogenic and angiogenic pathways, whereas, there was induction of pathways for immune system activation and production of PGF. The pattern of PGF-induced gene expression was similar in corpus luteum not destined for luteolysis (2X-non-regressed) after the first PGF pulse but was very distinct after the second PGF pulse. Thus, although the initial PGF pulse induced mRNA for many pathways, the second and later pulses of PGF appear to have set the distinct pattern of gene expression that result in luteolysis.

Induction of mRNA for chemokines and chemokine receptors by prostaglandin F2α is dependent upon stage of the porcine corpus luteum and intraluteal progesterone.

This study tested the hypotheses that prostaglandin (PG) F(2α) increases expression of genes related to recruitment of leukocytes in mature but not early corpus luteum (CL) and that insensitivity to PGF(2α) action in early CL is dependent on high intraluteal progesterone (P4) concentrations. Experiment 1 examined early (0.5 h) and late (10 h) in vivo effects of PGF(2α) on mature (d 17 of pseudopregnancy) and early (d 9) porcine CL. Real-time PCR was used to measure mRNA for chemokines (IL8, CXCL2, CCL2, CCL8, CCL4, CCL11) and chemokine receptors (CCR1, CCR2, CXCR2, CCR5). Western blotting was used to measure protein expression and phosphorylation of nuclear factor-κB proteins. Treatment with PGF(2α) for 10 h increased mRNA for almost all of these genes (all expect CXCL2 and CCL11) in d 17 CL but not d 9 CL. Treatment with PGF(2α) also led to greater phosphorylation of nuclear factor-κB-1A protein in d 17 than d 9 CL. Experiment 2 had a 2 × 2 factorial design with d 9 gilts treated or not treated with epostane (3ß-hydroxysteroid dehydrogenase inhibitor to suppress intraluteal P4) and treated or not treated with PGF(2α). Treatment with PGF(2α) (10 h) or epostane alone did not induce expression of any of these genes in d 9 CL. However, PGF(2α) + epostane increased expression of all of these genes except CCL11. In conclusion, PGF(2α) increases mRNA for chemokines and chemokine receptors in mature CL with similar PGF(2α) effects induced in early CL if intraluteal P4 is suppressed prior to PGF(2α) treatment.

Effect of decreasing intraluteal progesterone on sensitivity of the early porcine corpus luteum to the luteolytic actions of prostaglandin F2alpha.

Prostaglandin F2alpha (PGF) causes luteolysis of the pig corpus luteum (CL) only after Day 12 of the estrous cycle. Recent evidence indicates that progesterone (P4) may protect the CL from cell death. The present study tested the hypothesis that acute inhibition of P4 by treatment with epostane (EPO; 3betaHSD inhibitor) in CL lacking luteolytic capacity (Day 9 CL) will allow PGF to induce responses associated with luteolysis. Multiple PGF-induced responses were evaluated, including genes involved in production of PGF and estradiol-17beta, apoptosis (caspase 3), and transcription (FOSB). These responses are associated with PGF-induced luteolysis and do not normally occur in CL lacking luteolytic capacity. Animals on Day 7 after estrus were divided into four groups: 1) control (C), 2) PGF, 3) EPO, and 4) PGF plus EPO (PGF+EPO). Treatment with EPO (10 mg/kg) or vehicle was given every 12 h for 36 h. Treatment with PGF (25 mg) or vehicle was given at 38 h, and CL were collected from all animals at 48 h. Some CL from each animal were frozen in liquid nitrogen for mRNA and protein analysis. Remaining CL were incubated in media for 2 h for determination of P4 and PGF production. EPO dramatically decreased production of P4 by luteal tissue (ng/mg tissue) by 90% and 95% in EPO and PGF+EPO groups, respectively, compared to C (P < 0.01). Low production of PGF by luteal tissue was found in C, PGF, and EPO groups; however, treatment with PGF+EPO dramatically increased (782%) luteal PGF production. Similar to intraluteal PGF production, increased mRNA for cyclooxygenase 2 (PTGS2) and phospholipase A2 (group IB; PLA2G1B) was found in the PGF+EPO, but not in the EPO or PGF, group. Aromatase (CYP19A1) mRNA was not induced by PGF or EPO; however, PGF+EPO caused a more than 40-fold increase in CYP19A1 mRNA (P < 0.01). CASP3 mRNA was increased (P < 0.01) by EPO (3.4-fold) and by PGF (2.7-fold) but was most dramatically increased by PGF+EPO (5.3-fold), whereas caspase activity was only increased by PGF (1.5-fold) or PGF+EPO (2.2-fold). Thus, these data support the hypothesis that elimination of the protective effect of intraluteal P4 does not directly cause luteolysis of the early CL but allows PGF to induce luteolytic responses in CL lacking luteolytic capacity.

The role of luteinizing hormone in regulating gene expression during selection of a dominant follicle in cattle.

At approximately 8.5 mm in diameter, the future dominant follicle is "selected" for continued growth in cattle. In the present study, cows were treated with a gonadotropin-releasing hormone receptor antagonist, acyline, just before follicle selection (near 7.8 mm) to investigate the role of LH in changing mRNA concentrations during selection of a dominant follicle. The ovaries containing the expected dominant follicle (EDF; first largest follicle) and expected largest subordinate follicle (ESF) were removed after 12 or 24 h of treatment. Real-time PCR was used to determine mRNA concentrations. ELISA was used to measure testosterone and 17beta-estradiol (E(2)) and radioimmunoassay to measure androstenedione (A(4)) in follicular fluid. Concentrations of E(2) were greater in EDF than in ESF of untreated cows near the time of follicle selection (12 h) or at 12 h after selection (24 h). Testosterone, E(2), and A(4) were all dramatically decreased by acyline treatment at both times. In theca cells, acyline treatment reduced CYP17A1 (P450 17alpha) in EDF and STAR (steroidogenic acute regulatory protein) in both EDF and ESF but did not alter CYP11A1 (P450scc). In granulosa cells (GCs), LHCGR (luteinizing hormone [LH] receptor) was much greater in EDF than in ESF at both time of selection (739% greater) and 12 h after selection (2837% greater) and was decreased by acyline in EDF (87% decrease). The mRNA for CYP19A1 (cytochrome P450 aromatase) and PAPPA (pregnancy-associated plasma protein-A) tended to be greater in EDF than in ESF at follicle selection, and both mRNAs were much greater at 12 h after selection, with acyline significantly decreasing PAPPA mRNA after 24 h of treatment. The mRNA for FSHR (follicle-stimulating hormone receptor) was not different in EDF versus ESF and was not altered by acyline. Thus, induction of LHCGR mRNA in GCs is an early event during the follicle selection process, and surprisingly, expression of LHCGR mRNA is dependent on circulating LH. Production of follicular A(4), testosterone, and E(2) are also acutely related to LH but due to changes in expression of STAR and CYP17A1 in TC.

Distinct regulation by steroids of messenger RNAs for FSHR and CYP19A1 in bovine granulosa cells.

Steroidal regulation of gene expression in follicular cells is not completely defined. Granulosa cells from 5 mm bovine follicles were cultured and treated and steady-state mRNA levels determined for FSHR (follicle-stimulating hormone receptor) and CYP19A1 (aromatase). Cells were treated for 5 days with (0.1-300 ng/ml) 17beta-estradiol (E2), testosterone (T), or 5alpha-dihydrotestosterone (DHT). FSHR mRNA was increased by T and DHT but not E2. In contrast, CYP19A1 mRNA was induced by all doses of E2 but only high doses of T and DHT. Similarly, varying treatment duration (1-5 days) showed that FSHR was increased by T and DHT and CYP19A1 mRNA increased by E2 and T at all times. Synergism between steroid hormones and FSH or forskolin was also evaluated. FSH or E2 did not alter FSHR mRNA and did not enhance DHT stimulation of FSHR mRNA. In contrast, DHT alone had no effect on CYP19A1 mRNA but synergized with FSH plus E2 to increase CYP19A1 mRNA, probably due to induction of FSHR by DHT. Effects of E2 and T on CYP19A1 were blocked by ICI 182,780, indicating mediation by estrogen receptors. However, the specific androgen receptor antagonist bicalutamide did not block E2 or T effects on CYP19A1 but did block T and DHT stimulation of FSHR. Thus, FSHR is specifically regulated through androgen receptor, whereas CYP19A1 is regulated by multiple pathways, including estrogen receptors and cAMP/protein kinase A induced by FSHR activation in granulosa cells. These inter- and intracellular regulatory mechanisms may be critical for normal follicle growth and dominant follicle selection.