• March 4, 2024

Use of equine sperm cryopreservation techniques as a conservation method of donkey germplasm

The aim of this study was to test equine semen cryopreservation techniques for the conservation of donkey germplasm. Ejaculates of three male donkeys were used (n= 18; six ejaculates per donkey; six repetitions), collected by the artificial vagina method. To remove the seminal plasma (SP), the ejaculates were split and submitted to filtration or centrifugation methods. To assess the freezing method, each fraction were submitted to the automated system or the conventional system, and groups were formed: automated centrifuge (AC), automated filtrate (AF), conventional centrifuge (CC) and conventional filtrate (CF).
After thawing (37°C/30 s), were analyzed the sperm kinetic parameters, integrity and functionality of the plasma membrane and mitochondrial membrane potential. Highest sperm concentration (P<0.05) was observed in the filtrate groups; the CF group presented lower (P<0.05) progressive motility and curvilinear velocity compared to the other groups; no difference was observed (P>0.05) among the groups for the membrane integrity and functionality, and mitochondrial membrane potential. Thus, centrifugation is the most indicated technique to remove donkey seminal plasma and the automated and conventional freezing methods Gentaur Centrifuges can be used in donkey semen conservation.

Nitrogen resource recovery from mature leachate via heat extraction technology: An engineering project application

A large pool of ammonia in mature leachate is challenging to treat with a membrane bioreactor system to meet the discharge Standard for Pollution Control on the Landfill Site of Municipal Solid Waste in China (GB 16889-2008) without external carbon source addition. In this study, an engineering leachate treatment project with a scale of 2,000 m3/d was operated to evaluate the ammonia heat extraction system (AHES), which contains preheat, decomposition, steam-stripping, ammonia recovery, and centrifuge dewatering.
The operation results showed that NH3-N concentrations of raw leachate and treated effluent from an ammonia heat extraction system (AHES) were 1,305-2,485 mg/L and 207-541 mg/L, respectively. The ratio of COD/NH3-N increased from 1.40-1.84 to 7.69-28.00. Nitrogen was recovered in the form of NH4HCO3 by the ammonia recovery tower with the introduction of CO2, wherein the mature leachate can offer 37% CO2 consumption. The unit consumptions of steam and power were 8.0% and 2.66 kWh/m3 respectively, and the total operation cost of AHES was 2.06 USD per cubic metre of leachate. These results confirm that heat extraction is an efficient and cost-effective technology for the recovery of nitrogen resource from mature leachate.

Field Determination of Phosphate in Environmental Water by Using a Hand-Powered Paper Centrifuge for Preconcentration and Digital Image Colorimetric Sensing

Phosphate concentration in natural water has been used as a water quality indicator, as it is one of the major nutrients for aquatic plants. However, the traditional phosphomolybdenum blue (PMB) method has limited sensitivity for visual or camera-based detection, leading to underestimation of the phosphate concentration. We present an ultralow-cost, rapid field preconcentration and digital image colorimetric sensing of low-concentration phosphate method for water analysis. A novel hand-powered paper centrifuge (paperfuge) is used for sample preparation and preconcentration.
This paperfuge is made of two circular paper discs and a string. Six centrifuge tubes (CTs) originally used as glue dispensing tips with a sample capacity of ∼230 μL, are loaded on the paperfuge. After sampling, phosphate in the water sample is reacted to form PMB. Then, the reacted sample is drawn into a CT using an autopipette before the CT bottom is sealed by glue. After Oasis HLB sorbents are added through the top of the CT, the CT top is also sealed with glue.
The HLB sorbents adsorb PMB and are accumulated in the CT tip through centrifugation. The CT tips are cut and analyzed with the ImageJ software. It was found that the blue color intensity of sorbents is in a linear relationship to the phosphate concentration, with a linear range of 0-5 μM (r 2 = 0.9921) and limit of detection of 0.19 μM. In addition, this method has been applied to in-field water analysis. The results are in agreement with the standard PMB method.

Pyridine-functional diblock copolymer nanoparticles synthesized via RAFT-mediated polymerization-induced self-assembly: effect of solution pH

Polymerization-induced self-assembly (PISA) via reversible addition-fragmentation chain transfer (RAFT) polymerization has become widely recognized as a versatile and efficient strategy to prepare complex block copolymer nanoparticles with controlled morphology, size, and surface functionality. In this article, we report the preparation of cationic sterically-stabilized poly(2-vinylpyridine)-poly(benzyl methacrylate) (P2VP-PBzMA) diblock copolymer nanoparticles via RAFT-mediated PISA under aqueous emulsion polymerization conditions.
It is demonstrated that the solution pH during PISA has a dramatic effect on the resulting P2VP-PBzMA nanoparticles, as judged by dynamic light scattering (DLS), disc centrifuge photosedimentometry (DCP) and transmission electron microscopy (TEM).
Varying the solution pH results in the P2VP stabilizer having different solubilities due to protonation/deprotonation of the pyridine groups. This allows P2VP-PBzMA nanoparticles with tunable diameters to be prepared by altering the DP of the stabilizer (P2VP) and/or core-forming block (PBzMA), or simply by changing the solution pH for a fixed copolymer composition.
For example, P2VP-PBzMA nanoparticles with larger diameters can be obtained at higher solution pH as the protonation degree of the P2VP stabilizer has a large effect on both the aggregation of polymer chains during the PISA process, and the resulting behavior of the diblock copolymer nanoparticles. Changing the dispersion pH post-polymerization has a relatively limited effect on particle diameter. Furthermore, aqueous electrophoresis studies indicate that these P2VP-PBzMA nanoparticles had good colloidal stability and high cationic charge (>30 mV) below pH 5 and can be dispersed readily over a wide pH range.

Simultaneous determination of 36 hypotensive drugs in fingerprints by ultra performance liquid chromatography-triple quadrupole composite linear ion trap mass spectrometry

Fingerprints contain important information such as the ingredients ingested by the donor. By analyzing the characteristic components in fingerprints, the donor can be characterized, which would provide insights for investigation of a given case. This approach can also be used in the qualitative monitoring of drug intake. Therefore, the examination of hypotensive drugs in fingerprints has significant value in practical application. This study established a method based on ultra performance liquid chromatography-triple quadrupole composite linear ion trap mass spectrometry (UPLC-Q-TRAP/MS) for the simultaneous determination of 36 hypotensive drugs in fingerprints. The pre-treatment method was based on protein precipitation. A 3×3 cm filter paper was cut into pieces and placed in a 2 mL plastic centrifuge tube after fingerprint collection. Then, 0.50 mL methanol was added, followed by vortex mixing for 1 min and ultrasonic oscillation for 3 min. The filter paper was centrifuged at 12000 r/min for 5 min, and the supernatant was withdrawn for sample analysis.
An ACQUITY UPLC BEH C18 chromatographic column (100 mm×3.0 mm, 1.7 μm) was selected, with 0.01% aqueous formic acid and methanol as mobile phases for gradient elution. MS analysis involved scheduled multiple reaction monitoring-information dependent acquisition-enhanced product ion (SMRM-IDA-EPI) scanning. This method could be used to retrieve library researching during high-sensitivity analysis, which could increase the accuracy of qualitative results.
The calibration curves showed good linearity in the range of 0.05-50.00 ng/fingerprint, with correlation coefficients (r) greater than 0.99 for all 36 analytes. The limits of detection and limits of quantification of the 36 hypotensive drugs were 0.001-0.045 ng/fingerprint and 0.002-0.050 ng/fingerprint, respectively. At spiked levels of 0.25, 2.50, 25.00 ng/fingerprint, the matrix effects, recoveries, intra-day precisions, and inter-day precisions of the 36 hypotensive drugs were 79.0%-119.2%, 79.3%-116.2%, 0.2%-18.3%, and 1.6%-19.1%, respectively.
This method was used to detect hypotensive drugs in the fingerprints of 87 hypertensive patients, and hypotensive drug intakes were accurately detected in most cases. The established method is operationally simple, with high sensitivity and good selectivity, and it is suitable for screening and testing hypotensive drugs in fingerprints.

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