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Ammonia emission has attracted considerable attention concerning environmental protection in the world. About 33% of the atmospheric ammonia emission comes from farmland fertilization, so the monitoring of ammonia emissions from farmland is of great significance for controlling atmospheric ammonia pollution. In recent years, tuneable diode laser absorption spectroscopy (TDLAS) has been applied to the measurement of farmland ammonia emission; however, the spectral signal of the open optical path is susceptible to distortion due to temperature, noise, turbulence, etc. Meanwhile, the inversion of the distorted spectrum will result in abnormal concentration values that reduce the detection accuracy. This paper proposes a TDLAS spectral quality evaluation method based on dynamic time warping (DTW) algorithm and support vector data description (SVDD) algorithm. DTW algorithm was used to analyse the similarity between the measured spectrum and the reference spectrum, then SVDD algorithm was used to generate a classification model according to the results of DTW algorithm. DTWSVDD algorithm could effectively avoid obtaining wrong inversion concentration and improve the accuracy of concentration monitoring with identification accuracy of spectral quality reached more than 98%. Applied to the measurement of farmland ammonia emission in Fengqiu, Henan Province, China at the Agricultural Ecological Experiment Station of Chinese Academy of Sciences in November 2021, this method could effectively reduce the dispersion degree of measured concentration trend, as a result, which was better than that of the reported correlation coefficient method. DTW-SVDD algorithm should be of further research value for the application of the analysis of the Big Data of atmospheric composition detection based on grand-based optical remote sensing system. [ABSTRACT FROM AUTHOR]

The objective was to determine whether enucleated oocytes injected with frozen porcine first polar bodies (pPB1s) could be fertilized and developed into viable embryos in vitro. Metaphase II (MII) oocytes with pPB1s were frozen (vitrified) and stored for 2 mo. The pPB1s were isolated from thawed MII oocytes and injected into enucleated recipient oocytes by micromanipulation. All recipients injected with thawed pPB1s were fertilized by intracytoplasmic sperm injection (ICSI), and the resulting recombinant zygotes were incubated to assess their developmental competence in vitro. Furthermore, double-antibody immunohistochemistry was used to verify that the nucleus of the pPB1 participated in fertilization and supported embryonic development. Porcine embryos (2- to 8-cell stage) were obtained from the recombinants. The average in vitro cleavage rate of 2-, 4-, and 8-cell stage recombinant embryos was 25.3, 17.7, and 9.3% (P < 0.05), respectively. Chromosomes in the labeled pPB1 participated in the formation of the two blastomere nuclei of 2-cell stage embryos derived from recombinant oocytes. In conclusion, nuclear materials of frozen-thawed pPB1 supported oocyte fertilization and subsequent embryonic development, thereby providing a new way to use frozen PB1s for preservation and reproduction of mammals., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Chromatin condensation and germinal vesicle breakdown (GVBD) in mouse cumulus free oocytes was blocked by 2 mmol/L of 6-DMAP in vitro. Mouse oocytes (M II) from superovulated female mice could be activated by 6-DMAP. The activation rates of oocytes (18-19 h post hCG) cultured in CZB medium with 2 mmol/L 6-DMAP for 0.5, 1, 2, 3 h, were 26.1%, 75.2%, 75.8%, 77.3% respectively (Table 1), and the cleavage rates of these activated oocytes were 88.2%, 73.2%, 67.0%, 58.4% respectively (Table 2). The parthenogenetic activation type induced by 6-DMAP was different from that produced by ethanol (Fig.1).

Mitochondrial reorganization during meiotic maturation and parthenogenetic activation was studied in mouse oocytes using a laser scanning confocal microscope and a transmission electron microscope. Mitochondria were mainly distributed perinuclearly in the germinal vesicle (GV) stage oocytes and were dispersed throughout ooplasm after germinal vesicle breakdown (GVBD), except for a slightly higher occurrence in one hemisphere of oocytes, from which the first polar body (PbI) would become extruded. Mitochondria reaggregated around the metaphae II (MII) spindle and pronuclear region after alcohol activatation at the MII stage. The mitochondrial distribution may correspond to the Ca(2+) changes during meiotic maturation and parthenogenetic activation.

Aim: To study the mechanism of oocyte activation in mammals., Methods: Mouse oocytes arrested at metaphase of the second meiotic division were loaded with Fura 2-AM and then activated with ethanol, calcimycin, electric pulse or fertilization. Intracellular free Ca2+ during activation were measured by Spex AR-CM-MIC cation system. Cortical granule exocytosis after activation was detected under electron microscopy., Results: Sperm penetration initiated a long lasting Ca2+ oscillation in Ca2+ containing IVF medium in mouse ova. The Ca2+ oscillation lasted for over 3-4 h until the ova developed to pronuclear stage. The Ca2+ oscillated faster as extracellular Ca2+ concentration was increased from normal 1.7 mmol.L-1 to 5.0 mmol.L-1 and ceased to oscillate when extracellular Ca2+ was removed. The ova resumed Ca2+ oscillation after transferred back to IVF (Ca2+ 1.7 mmol.L-1). The ova which exhibited Ca2+ oscillation all extruded second polar body and formed pronuclei. Suppression the Ca2+ oscillation by intracellular injection of egtazic acid (2-10 pL, 200 mumol.L-1) blocked the activation of oocytes. Heparin (100 mumol.L-1) injection failed to prevent the Ca2+ oscillation. In artificial activation, ethanol, calcimycin, and a single electric pulse usually induced a monotonic Ca2+ rise and resulted in the activation only in older oocytes (> 18 h after CG injection). Activation of freshly ovulated oocytes required multiple intracellular Ca2+ increases induced by repetitive electric pulses. Artificial activation elicited the similar cortical granule exocytosis in oocytes as occurring at fertilization. Suppression of the intracellular Ca2+ elevation by introduction of egtazic acid into the oocytes blocked the activation process., Conclusions: The increase of intracellular free Ca2+ is the primary signal responsible for the initiation of oocyte activation but there are distinct differences between fertilization and artificial activation both in Ca2+ change patterns and Ca2+ sources. Young oocytes require oscillatory Ca2+ signals for activation.

The effect and mechanism of EGF and progesterone on the meiotic maturation resumption were studied. The results showed that progesterone regulated the action of EGF. The inhibitor of 3 beta-HSD, the key enzyme of progesterone synthesis, Epostane could not only prevent the effect of EGF, but inhibit the progesterone production promoted by EGF in cultured ovary granulosa cells as well. On the other hand, heparin could lessen the function of EGF and progesterone, which implies that calcium participated the effect of EGF and progesterone. Fura-2 was used to indicate the changes of intracellular calcium level in single cumulus cell. It is found that a calcium wave appeared in cumulus cell when EGF or progesterone acted. But Ca2+ level returned to normal immediately after progesterone was added, and Ca2+ in the cumulus cell still remained higher level after EGF acted. This results indicated that calcium in cumulus cell adjusted the action of EGF and progesterone.

The effect of purine on the meiosis of mouse oocytes was studied. When adenine and benzyladenine, a similar substance of adenine, was microinjected into the germinal vesicle of oocytes, the meiotic maturation of oocytes could be obviously arrested depending on the dosage of injection. The stimulator of adenylate cyclase, NaF, could enhance the arrest effect of adenine. It indicated that cAMP played a important role in mouse oocytes meiotic resumption. Hypoxanthine and adenine had different effects in different medium. Hypoxanthine could obviously arrest the maturation of mouse cumulus oocyte complex (COC) and cumulus denuded oocytes (DO) in DMEM or EMEM. But only in DMEM adenine could obviously arrest the maturation of COC, and had co-effect with hypoxanthine in DMEM. They could almost arrest the maturation of COC and DO. Adenine in EMEM had little arrest effect on maturation of COC and DO. The different effects may result from the different adenine absorption by oocyte in DMEM and EMEM. Adding glutamine into EMEM could enhance the arrest effect of adenine to COC and DO.

Oocytes collected 18-19 h after HCG injection were stimulated with 7-8% ethanol or electrical pulses (1.7 KV/cm field strength, 80-100 microseconds duration, 3-4 times, 5-6 min interval). The parthenogenetic embryos derived from the above-mentioned methods developed to blastocyst stage just like those developed from fertilized eggs. Mouse oocytes were rather sensitive to ethanol stimulation. More than 95% of the treated oocytes were activated after stimulation of 7-8% ethanol for 5 min. Multiple electrical stimulations induced higher activation percentages of oocytes than only single electrical stimulation (71.5% vs. 63.6%). Intact oocytes were loaded with fluorescent Ca2+ indicator fura-2 and intracellular free calcium changes during artificial activation were measured by fluorescence detector. The results showed that ethanol could induce repetitive transient Ca2+ concentration increase in activated oocytes. Single electrical stimulation only induced single free calcium concentration elevation in oocyte while multiple electrical pulses could induce repetitive Ca2+ increase (each electrical pulse elicited the corresponding Ca2+ concentration peak). The pronuclei were not observed in the oocytes which had not exhibited calcium concentration rise during activation. Apart from electrical stimulation parameter, sufficient amount of Ca2+ in electric medium was crucial to mouse oocyte activation when stimulated with electrical pulses. The oocytes were hardly activated by electrical stimulations in a medium without Ca2+ even with longer pulse duration and the intracellular free calcium concentration in the oocytes showed no elevation. This indicates that the inflow of extracellular Ca2+ from tiny pores across the oocyte membrane caused by electrical stimulation is the main source of intracellular free calcium increase.(ABSTRACT TRUNCATED AT 250 WORDS)