41 Effect of vitrification on DNA integrity of human spermatozoa

2020 ◽  
Vol 32 (2) ◽  
pp. 146
Author(s):  
Y. Toishibekov ◽  
B. Shalekenov ◽  
Y. Assanova ◽  
S. Shalekenov ◽  
Y. Kuandykov ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Liquid Nitrogen ◽  
Dna Fragmentation ◽  
Room Temperature ◽  
Human Spermatozoa ◽  
Basic Medium ◽  
Sperm Suspension ◽  
Human Tubal Fluid

The aim of our study was to investigate the effect of vitrification with sucrose on swim-up-prepared human spermatozoa in comparison with standard, conventional manual freezing with permeable cryoprotectants. After informed consent, 35 ejaculates were obtained from 35 patients with normozoospermia who were patients of a fertility clinic. All specimens used for this study had fulfilled the following quality criteria for spermatozoa concentration and motility on IVOS (Hamilton Thorne). Semen analysis was performed according to published guidelines of the World Health Organization (WHO Laboratory Manual for the Examination and Processing of Human Semen, 2010). After swim-up, each sample was centrifuged, resuspended with the basic medium (human tubal fluid + 1% human serum albumin) to achieve a concentration of 5×106 spermatozoa/mL, and finally aliquoted into two equal subsamples. Each of these aliquots was assigned to one of two groups: group 1 included conventionally cryopreserved spermatozoa and group 2 included spermatozoa that were vitrified. For conventional cryopreservation, freezing media (15% (vol/vol) glycerol, 20% (vol/vol) egg yolk) and citrate was added to the washed spermatozoa in a 1:2 ratio. The sperm suspension was aspirated into 0.5-mL straws (CryoBioSystem). Subsequent to the room-temperature incubation for 10min, straws were placed horizontally in the vapour phase for 15min and then submerged into liquid nitrogen. For thawing, cryopreserved straws were immersed in water (23°C) for 5min. For preparation of vitrification solution, the basic medium (human tubal fluid + 1% human serum albumin) was diluted 1:1 with 0.5M sucrose. Immediately after processing, the sperm suspension was diluted in a 1:1 ratio with the vitrification solution to reach a final sucrose concentration of 0.25M. The vitrification and sperm solution (300μL) were aspirated into the straws 0.5mL. Straws were then left at room temperature (20-21°C) for 10min and subsequently submerged horizontally into the liquid nitrogen (Isachenko et al. 2012 J. Androl. 33, 462-468; https://doi.org/10.2164/jandrol.111.013789) and stored similarly to the conventionally cryopreserved straws. To thaw, vitrified straws were immersed in a water bath (42°C) for 20s. The DNA fragmentation was analysed using the APO DIRECT kit (BD PharmingenTM). The cells were stained according to the manufacturer's protocol, followed by flow cytometry analysis CyFlow (Sysmex-Partec). An analysis of variance with a significance of 0.05 for nonparametric statistical analysis to establish differences between groups was used. In our study, no statistically significant differences were observed in the total motility, progressive motility, or velocity parameters of spermatozoa (P>0.05) post-thawing. Also, higher percentages of DNA fragmentation (35.1±8.1% vs. 20.1±6.8%; P<0.05) were found in spermatozoa cryopreserved by means of vitrification with sucrose compared with conventional cryopreservation. Therefore, these methods are comparable and either can be implemented for the storage of spermatozoa to be used for future assisted-reproduction-technology procedures. Vitrification of human spermatozoa provides a simpler, faster, more cost-effective alternative to conventional cryopreservation methods.

Laser immunonephelometry for routine quantification of urinary albumin excretion.

1987 ◽  
Vol 33 (2) ◽  
pp. 209-213 ◽  
Author(s):  
M Marre ◽  
J P Claudel ◽  
P Ciret ◽  
N Luis ◽  
L Suarez ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Room Temperature ◽  
Normal Values ◽  
Urinary Albumin Excretion ◽  
Urinary Albumin ◽  
Albumin Excretion ◽  
Large Numbers ◽  
Time Of Collection

Abstract We describe a laser-immunonephelometric method for quantifying urinary albumin excretion (UAE) in large numbers of samples. For a 150-microL sample incubated at room temperature for 45 min with 40 microL of antiserum specific for human serum albumin, the assay range for albumin was 0.34 to 43.0 mg/L. For samples analyzed undiluted and diluted 10-fold, the range of measurable albumin was from 0.34 to 430.0 mg/L. With an automated version of this method, one can assay 240 samples per hour. Intra- and interassay CVs were less than 6% and 9%, respectively. Measurements by this method (y) correlated well with those obtained by a RIA method (x): y = 1.00x + 0.163 mg/L (n = 233; r = 0.996). The day-to-day CV for UAE was determined for three consecutive determinations done on each of 60 controls according to the time of collection and in 212 diabetics according to the amount of 24-h UAE. For controls, UAE was 8.0 +/- 8.1 mg/24 h (mean +/- SD), CV 44 +/- 23%. The CV was similar for diurnal (50 +/- 28%) and overnight (58 +/- 32%) collections from controls; and for diabetics with normal values for UAE: 35 +/- 32%, with slight albuminuria (25-300 mg/24 h):37 +/- 28%, or with macroalbuminuria (greater than 300 mg/24 h): 47 +/- 42%.

Z-Acrylonitrile Derivatives: Improved Synthesis, X-ray Structure, and Interaction with Human Serum Albumin

2020 ◽  
Vol 16 (8) ◽  
pp. 1149-1160
Author(s):  
Mehtab Parveen ◽  
Afroz Aslam ◽  
Shahab A.A. Nami ◽  
Musheer Ahmad
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Efficient Method ◽  
Zinc Chloride ◽  
Room Temperature ◽  
Internal Standard ◽  
Solid Catalyst ◽  
Heterocyclic Aldehydes ◽  
Aromatic Heterocyclic

Aim and Objective: In the synthesis of heterocyclic compounds, acrylonitrile derivatives are the most important and appropriate precursors. These compounds are the most important intermediates and subunits for the enhancement of molecules having pharmaceutical or biological interests. Nitrogen-containing compounds have received extensive consideration in the literature over the years. Materials and Methods: A facile, economic and efficient method has been developed for the synthesis of acrylonitrile derivatives using p-nitrophenylacetonitrile and aromatic/heterocyclic aldehydes in the presence of zinc chloride at room temperature. Spectroscopic data were obtained using the following instruments: Fourier transform infrared spectra (KBr discs, 4000-400 cm-1) by Shimadzu IR-408 Perkin-Elmer 1800 instrument; 1H NMR and 13C NMR spectra by Bruker Avance-II 400 MHz using DMSO-d6 as a solvent containing TMS as the internal standard. Results: To continue our ongoing studies to synthesize heterocyclic and pharmaceutical compounds by mild, facile and efficient protocols, herein we wish to report our experimental results on the synthesis of acrylonitrile derivatives, using various aromatic/heterocyclic aldehydes and p-nitrophenylacetonitrile in the presence of zinc chloride in ethanolic media at room temperature. Some of the new compounds were tested for their human serum albumin activity (HSA) while a study of interaction with HSA protein was performed for compounds 3a and 3b. The results show that compound 3b binds tightly to HSA as compared to compound 3a. Conclusion: It can be concluded that acrylonitrile derivatives can be synthesized by an efficient method via the reaction of p-nitrophenylacetonitrile with aromatic/heterocyclic aldehydes by the use of zinc chloride as an effective solid catalyst. The remarkable features of this procedure include excellent yields (90-95%), short reaction period (30 min.), moderate reaction environment, easy workup procedure and managing of the catalyst. This method may find a wide significance in organic synthesis for the synthesis of the Z-acrylonitrile.

Comparative Survival of Human Spermatozoa in Media Supplemented With Polyvinyl Alcohol or Human Serum Albumin

2000 ◽  
Vol 74 (3) ◽  
pp. S246 ◽  
Author(s):  
M.H Javed ◽  
M Geisman ◽  
M.A Shaikh ◽  
C Ruberto ◽  
A.P Del Valle
Keyword(s):  
Human Serum Albumin ◽  
Polyvinyl Alcohol ◽  
Serum Albumin ◽  
Human Serum ◽  
Human Spermatozoa

II. Lokalisation der Placenta mit 113Inm- Human-Serum-Albumin

1969 ◽  
Vol 08 (01) ◽  
pp. 15-21 ◽  
Author(s):  
K. E. Scheer ◽  
J. Heep ◽  
W. Maier-Borst ◽  
W. J. Lorenz ◽  
H. Sinn ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Vena Cava ◽  
Placenta Praevia ◽  
Wird Eine

ZusammenfassungNach tierexperimentellen Voruntersuchungen wurde die Placentographie mit trägerfreiem 113Inm -HSA als klinische Methode eingeführt. Vor Amniocentesen und bei Verdacht auf Placenta praevia werden Placentographien geschrieben. Den Schwangeren wird eine Aktivität von 500 μCi in die Cubitalvene injiziert. Die der Aktivität entsprechende Indiummenge ist kleiner als 0,1 ng. Die fetale Strahlenbelastung liegt unter lOmrad. Bei Anwendung von 113Inm-HSA entfällt eine Blockade der mütterlichen und fetalen Schilddrüsen. Die genaue Abgrenzung einer Placenta praevia wird nicht durch eine Blasenaktivität beeinträchtigt.Es wurden bisher 19 Placentalokalisationen durchgeführt. In allen Fällen konnte der Placentasitz eindeutig festgestellt werden. Bedingt durch die lange Liegezeit beim Aufnehmen eines Szintigramms kam es in zwei Fällen zu einem Vena-Cava-Kompressions-Syndrom. Zur Verhinderung dieser klinischen Zwischenfälle werden inzwischen Placentographien mit der Anger-Kamera aufgenommen. Mit Hilfe des divergierenden Kollimators konnte der gesamte Abdominalbereich erfaßt werden. Die Aufnahmezeit konnte auf 7 — 10 Minuten verkürzt werden. Die intravenöse injizierte Aktivität betrug bei dieser Methode ebenfalls 500 μCi. Der diagnostische Aussagewert der Kamerabilder ist szintigraphischen Aufnahmen gleichwertig.

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