scholarly journals Effects of diadenosine polyphosphates and seminal fluid vesicles on rabbit sperm cells

Reproduction ◽  
2003 ◽  
pp. 827-835 ◽  
Author(s):  
A Minelli ◽  
L Liguori ◽  
I Bellezza ◽  
T Renieri ◽  
C Castellini
Keyword(s):  
High Performance ◽  
Acrosome Reaction ◽  
Seminal Fluid ◽  
Membrane Vesicles ◽  
Fluorescein Isothiocyanate ◽  
Plasma Electron ◽  
Sperm Cells ◽  
Experimental Conditions ◽  
Diadenosine Polyphosphates ◽  
Rabbit Spermatozoa

Membrane vesicles were isolated from rabbit seminal plasma. Electron microscopy analyses showed the presence of numerous small, round vesicles with a diameter of about 70 nm. Determination of enzyme activities was carried out by high performance liquid chromatography and showed that the vesicles can degrade the diadenosine polyphosphates (ApnA), Ap3A and Ap4A and ATP and ADP, but not AMP. Studies of the degradation of diadenosine compounds by the vesicles present in seminal fluid showed an increasing production of AMP as the by-product and a time-dependent generation of dephosphorylated products consistent with the presence of ecto-ATP diphosphophosphatase (ecto-apyrase). In the presence of rabbit spermatozoa, AMP did not accumulate because 5'nucleotidase and adenosine deaminase, present at the surface of sperm cells, transformed AMP into adenosine and inosine. The effects of seminal fluid vesicles and diadenosine compounds on the acquisition of fertilizing capacity by rabbit spermatozoa were evaluated by Pisum sativum agglutinin fluorescein isothiocyanate conjugated staining. The results obtained with uncapacitated spermatozoa showed that the capacitating effector BSA could be substituted efficiently by the addition of diadenosine compounds and vesicles previously incubated for 2 h to the capacitative medium. Under these experimental conditions, the spontaneous acrosome reaction rate was not increased. Capacitated rabbit spermatozoa did not undergo acrosome reaction when l-alpha-lysophosphatidylcholine was substituted by diadenosine compounds previously incubated with vesicles. In conclusion, this study has shown that rabbit seminal fluid vesicles can degrade diadenosine compounds to AMP and that the addition of the vesicles and diadenosine compounds to uncapacitated rabbit spermatozoa favours the acquisition of the fertilizing capacity.

Micro-Thermal Field-Flow Fractionation

2002 ◽  
Vol 67 (11) ◽  
pp. 1596-1608 ◽  
Author(s):  
Josef Janča
Keyword(s):  
High Performance ◽  
Thermal Field ◽  
Temperature Drop ◽  
Point Of View ◽  
Constant Field ◽  
Field Flow Fractionation ◽  
Operational Mode ◽  
Experimental Conditions ◽  
Standard Size ◽  
Flow Fractionation

The effect of miniaturization of the separation channel on the performance of thermal field-flow fractionation (TFFF) is substantiated theoretically. The experiments carried out under carefully chosen experimental conditions proved the high performance of the separation of polymers within an extended range of molar masses from relatively low up to ultrahigh-molar-mass (UHMM) samples. The new micro-TFFF allows to achieve high resolution when applying constant field force operation, it makes easy the programming of the temperature drop which is an advantageous operational mode from the point of view of the time of analysis, and it extends considerably the range of perfectly controlled temperature of the cold wall due to a substantial decrease in the heat energy flux compared with standard size channels.

scholarly journals How Relevant Is It to Use Mineral Proxies to Mimic the Atmospheric Reactivity of Natural Dust Samples? A Reactivity Study Using SO2 as Probe Molecule

Minerals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 282
Author(s):  
Darya Urupina ◽  
Manolis N. Romanias ◽  
Frederic Thevenet
Keyword(s):  
High Performance ◽  
Real Life ◽  
Complex Mixtures ◽  
Experimental Conditions ◽  
The Real ◽  
Atmospheric Processes ◽  
Real Effects ◽  
Mineral Aerosols ◽  
Life Impact ◽  
Reactivity Study

The experimental investigation of heterogeneous atmospheric processes involving mineral aerosols is extensively performed in the literature using proxy materials. In this work we questioned the validity of using proxies such as Fe2O3, FeOOH, Al2O3, MgO, CaO, TiO2, MnO2, SiO2, and CaCO3 to represent the behavior of complex mixtures of minerals, such as natural desert and volcanic dusts. Five volcanic dusts and three desert dusts were compared to a number of metal oxides, commonly used in the literature to mimic the behavior of desert dusts in the ability to form sulfites and sulfates on the surface exposed to SO2 gas. First, all samples were aged at room temperature, atmospheric pressure, under controlled experimental conditions of 175 ppm SO2 for 1 h under 30% of relative humidity. Second, they were extracted with 1% formalin and analyzed by High-Performance Liquid Chromatography (HPLC) to quantify and compare the amount of sulfites and sulfates formed on their surfaces. It was evidenced that under the experimental conditions of this study neither one selected pure oxide nor a mixture of oxides can adequately typify the behavior of complex mixtures of natural minerals. Therefore, to evaluate the real-life impact of natural dust on atmospheric processes it is of vital importance to work directly with the natural samples, both to observe the real effects of desert and volcanic dusts and to evaluate the relevancy of proposed proxies.

Determination of the Isoflavone Genistein in Soybeans by High-Performance Liquid Chromatography Following Cloud Point Extraction

2012 ◽  
Vol 95 (3) ◽  
pp. 845-849 ◽  
Author(s):  
Mohammad Mirzaei ◽  
Atieh Karimi Naeini ◽  
Mansoureh Behzadi
Keyword(s):  
Cloud Point ◽  
High Performance ◽  
Cloud Point Extraction ◽  
Uv Detector ◽  
Experimental Conditions ◽  
Solid Ratio ◽  
Assisted Extraction ◽  
Ultrasonic Assisted ◽  
Replicate Measurements

Abstract A simple and sensitive method has been developed for preconcentration and determination of genistein in soybeans. This method is based on cloud point extraction (CPE) of genistein from soybeans using ethylene glycol monoalkyl ether (Genapol X-080) as a nonionic surfactant. The concentration of extracted genistein was determined by HPLC with a UV detector. Optimum experimental conditions were established. With 5% Genapol X-080 (v/v), a liquid/solid ratio of 25:1 mL/g, and ultrasonic-assisted extraction at 40°C for 45 min, the extraction percentage of genistein reached its highest value. The preconcentration factor for genistein was about 16.5. The RSD for seven replicate measurements and the LOD were ±4.45% and 15.0 ng/mL, respectively. CPE is simple, inexpensive, and suitable for extraction of genistein from soybean. It uses environmentally friendly surfactants and offers a convenient alternative to more conventional extraction systems.

scholarly journals Albumin binding of unconjugated [3H]bilirubin and its uptake by rat liver basolateral plasma membrane vesicles

1996 ◽  
Vol 316 (3) ◽  
pp. 999-1004 ◽  
Author(s):  
Lorella PASCOLO ◽  
Savino DEL VECCHIO ◽  
Ronald K. KOEHLER ◽  
J. Enrique BAYON ◽  
Cecile C. WEBSTER ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Rat Liver ◽  
Basolateral Membrane ◽  
Membrane Vesicles ◽  
Plasma Membrane Vesicles ◽  
Experimental Conditions ◽  
Saturation Kinetics ◽  
Basolateral Plasma Membrane ◽  
Component C ◽  
Uptake Studies

Using highly purified unconjugated [3H]bilirubin (UCB), we measured UCB binding to delipidated human serum albumin (HSA) and its uptake by basolateral rat liver plasma membrane vesicles, in both the absence and presence of an inside-positive membrane potential. Free UCB concentrations ([Bf]) were calculated from UCB–HSA affinity constants (K´f), determined by five cycles of ultrafiltration through a Centricon-10 device (Amicon) of the same solutions used in the uptake studies. At HSA concentrations from 12 to 380 μM, K´f (litre/mol) was inversely related to [HSA], irrespective of the [Bt]/[HSA] ratio. K´f was 2.066×106+(3.258×108/[HSA]). When 50 mM KCl was iso-osmotically substituted for sucrose, the K´f value was significantly lower {2.077×106+(1.099×108/[HSA])}. The transport occurred into an osmotic-sensitive space. Below saturation ([Bf] ⩽ 65 nM), both electroneutral and electrogenic components followed saturation kinetics with respect to [Bf], with Km values of 28±7 and 57±8 nM respectively (mean±S.D., n = 3, P < 0.001). The Vmax was greater for the electrogenic than for the electroneutral component (112±12 versus 45±4 pmol of UCB·mg-1 of protein·15 s-1, P < 0.001). Sulphobromophthalein trans-stimulated both electrogenic (61%) and electroneutral (72%) UCB uptake. These data indicate that: (a) as [HSA] increases, K´f decreases, thus increasing the concentration of free UCB. This may account for much of the enhanced hepatocytic uptake of organic anions observed with increasing [HSA]. (b) UCB is taken up at the basolateral membrane of the hepatocyte by two systems with Km values within the range of physiological free UCB levels in plasma. The electrogenic component shows a lower affinity and a higher capacity than the electroneutral component. (c) It is important to calculate the actual [Bf] using a K´f value determined under the same experimental conditions (medium and [HSA]) used for the uptake studies.

scholarly journals Creep Behaviour and Microstructural Characterization of VAT 36 and VAT 32 Superalloys

Metals ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 877 ◽  
Author(s):  
Vagner Gobbi ◽  
Silvio Gobbi ◽  
Danieli Reis ◽  
Jorge Ferreira ◽  
José Araújo ◽  
...  
Keyword(s):  
Failure Analysis ◽  
Creep Resistance ◽  
High Performance ◽  
Creep Behaviour ◽  
Ductile Failure ◽  
Microstructural Characterization ◽  
Dislocation Slip ◽  
Creep Tests ◽  
Experimental Conditions ◽  
Wide Range

Superalloys are used primarily for the aerospace, automotive, and petrochemical industries. These applications require materials with high creep resistance. In this work, evaluation of creep resistance and microstructural characterization were carried out at two new nickel intermediate content alloys for application in aerospace industry and in high performance valves for automotive applications (alloys VAT 32 and VAT 36). The alloys are based on a high nickel chromium austenitic matrix with dispersion of intermetallic L12 and phases containing different (Nb,Ti)C carbides. Creep tests were performed at constant load, in the temperature range of 675–750 °C and stress range of 500–600 MPa. Microstructural characterization and failure analysis of fractured surfaces of crept samples were carried out with optical and scanning electron microscopy with EDS. Phases were identified by Rietveld refinement. The results showed that the superalloy VAT 32 has higher creep resistance than the VAT 36. The superior creep resistance of the alloy VAT 32 is related to its higher fraction of carbides (Nb,Ti)C and intermetallic L12 provided by the amount of carbon, titanium, and niobium in its chemical composition and subsequent heat treatment. During creep deformation these precipitates produce anchoring effect of grain boundaries, hindering relative slide between grains and therefore inhibiting crack formation. These volume defects act also as obstacles to dislocation slip and climb, decreasing the creep rate. Failure analysis of surface fractures of crept samples showed intergranular failure mechanism at crack origin for both alloys VAT 36 and VAT 32. Intergranular fracture involves nucleation, growth, and subsequent binding of voids. The final fractured portion showed transgranular ductile failure, with dimples of different shapes, generated by the formation and coalescence of microcavities with dissimilar shape and sizes. The occurrence of a given creep mechanism depends on the test conditions. At creep tests of VAT 32 and VAT 36, for lower stresses and higher temperature, possible dislocation climb over carbides and precipitates would prevail. For higher stresses and intermediate temperatures shear mechanisms involving stacking faults presumably occur over a wide range of experimental conditions.

scholarly journals Concentrations of Gatifloxacin in Plasma and Urine and Penetration into Prostatic and Seminal Fluid, Ejaculate, and Sperm Cells after Single Oral Administrations of 400 Milligrams to Volunteers

2001 ◽  
Vol 45 (1) ◽  
pp. 293-297 ◽  
Author(s):  
Christoph K. Naber ◽  
Michaela Steghafner ◽  
Martina Kinzig-Schippers ◽  
Christian Sauber ◽  
Fritz Sörgel ◽  
...  
Keyword(s):  
Maximum Concentration ◽  
Seminal Fluid ◽  
Plasma Concentrations ◽  
Sperm Cells ◽  
Chronic Bacterial Prostatitis ◽  
Time Curve ◽  
Chromatography Method ◽  
Simultaneous Application ◽  
Bacterial Prostatitis ◽  
Antibacterial Spectrum

ABSTRACT Gatifloxacin (GTX), a new fluoroquinolone with extended antibacterial activity, is an interesting candidate for the treatment of chronic bacterial prostatitis (CBP). Besides the antibacterial spectrum, the concentrations in the target tissues and fluids are crucial for the treatment of CBP. Thus, it was of interest to investigate its penetration into prostatic and seminal fluid. GTX concentrations in plasma, urine, ejaculate, prostatic and seminal fluid, and sperm cells were determined by a high-performance liquid chromatography method after oral intake of a single 400-mg dose in 10 male Caucasian volunteers in the fasting state. Simultaneous application of the renal contrast agent iohexol was used to estimate the maximal possible contamination of ejaculate and prostatic and seminal fluid by urine. GTX was well tolerated. The means (standard deviations) for the following parameters were as indicated: time to maximum concentration of drug in serum, 1.66 (0.91) h; maximum concentration of drug in serum, 2.90 (0.39) μg/ml; area under the concentration-time curve from 0 to 24 h, 25.65 μg · h/ml; and half life, 7.2 (0.90) h. Within 12 h about 50% of the drug was excreted unchanged into the urine. The mean renal clearance was 169 ml/min. The gatifloxacin concentrations in ejaculate, seminal fluid, and prostatic fluid were in the range of the corresponding plasma concentrations which were 1.92 (0.27) μg/ml at approximately the same time point (4 h after drug intake). The concentrations in sperm cells (0.195, 0.076, and 0.011 μg/ml) could be determined in three subjects. The good penetration into prostatic and seminal fluid, the good tolerance, and the previously reported broad antibacterial spectrum suggest that GTX may be a good alternative for the treatment of chronic bacterial prostatitis. Clinical studies should be performed to confirm this assumption.

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