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Sperm Class Analyzer® articles:
 First Evaluation of Human Sperm Quality in Various Geographic Regions of Switzerland
Crausaz, Michel; Vargas, Josefina; Parapanov, Roumen; Chollet, Yves; Wisard, Marc; Stettler, Eric; Senn, Alfred; Germonda, Marc
A decline in human sperm quality and quantity has been reported in numerous Western countries. This
observation was also accompanied by an increase in urogenital malformations. The need for epidemiological
studies dealing with unbiased populations in order to understand the causes of these observations is obvious.
In Switzerland, the large majority of young men are asked to attend a military camp to be drafted into the army.
A few weeks before this camp, conscripts were contacted and invited to participate in a large national study on
semen quality. The participation was totally voluntary and anonymous. From September 2005 to June 2007, 770
volunteers filled out a questionnaire, underwent a clinical examination and provided sperm, blood and urine samples.
Using self-rated health assessments, the observed cohort could be considered as healthy and no testicular
cancer was found. Moreover, the testicular volumes, measured using Prader’s orchidometry and ultrasonography,
were comparable to those already published for young male populations. The median sperm concentration was
47 x 106 /ml, which is close to the concentration reported in Denmark, known to have the highest incidence of
testicular cancer in Europe. Statistically significant differences were observed between regions with a lower sperm
concentration for men residing in the Alps (43 x 106 /ml) and in the Zürich area (36 x 106 /ml) compared to men
from West Plateau (54 x 106 /ml) and from the Jura (54 x 106 /ml). Such a regional discrepancy could be related to
environmental factors, including endocrine disruptors. In order to confirm such regional differences more volunteers
from the already studied regions should be studied and other parts of the country should be investigated.
The rather low sperm concentration of Swiss young volunteers should be considered as a national health issue
and investigated further.
CHIMIA International Journal for Chemistry, Volume 62, Number 5, May 2008 , pp. 395-400(6)
10.2533/chimia.2008.395
Expression and localization of δ, κ and µ opioid receptors in human spermatozoa and implications for sperm motility
Ekaitz Agirregoitia, Asier Valdivia, Arkaitz Carracedo, Luis Casis, Javier Gil, Nerea Subiran, Carmen Ochoa and Jon Irazusta
CONTEXT: Endogenous opioid peptides signal through δ, κ and µ-opioid receptors.
Some of these peptides such as endorphins and enkephalins are present in the male
reproductive tract, but the presence of the corresponding receptors in human sperm cells
has not yet been reported.
OBJECTIVE: To study the expression and localization of δ, κ and µ-opioid receptors
on human spermatozoa and the implication in sperm motility.
METHODS: The expression of receptors was studied by RT-PCR, Western blot and
immunofluorescence techniques. We evaluated the effects of activation of each opioid
receptor by specific agonist and antagonist.
RESULTS: Human spermatozoa express δ, κ and µ-opioid receptors. These receptors
were located in different parts of the head, in the middle region and in the tail of the
sperm. Progressive motility of spermatozoa, an important parameter to evaluate male
fertility, was found to be significantly reduced after incubation with the µ-receptor
agonist morphine, whereas this effect was antagonized in the presence of the
corresponding antagonist, naloxone. The δ receptor antagonist naltrindole significantly
reduced progressive motility immediately after its addition. However, the δ receptor
agonist DPDPE had no significant effect. Finally, neither the κ receptor agonist U50488
nor its antagonist nor-binaltorphimine significantly affected the progressive motility of
human spermatozoa.
CONCLUSION: We report for first time the presence of functional δ-, κ- and µ-opioid
receptors in human sperm membranes. These findings are indicative of a role for the
opioid system in the regulation of sperm physiology.
J Clin Endocrin Metabol. 2006;91:4969–4975.
10.1210/jc.2006-0599
Ureaplasma urealyticum reduces motility and induces membrane alterations in human spermatozoa
Nunez-Calonge R, Caballero P, Redondo C, Baquero F, Martinez-Ferrer M, Meseguer MA.
Department of Gynecology, Ramon y Cajal Hospital, Institute Nacional of Health, Madrid, Spain.
The in-vitro effects of several concentrations of Ureaplasma urealyticum on the motility, membrane integrity and morphology of washed spermatozoa from healthy donors were studied. A significant reduction in sperm motility and signs of membrane alteration, directly related to U.urealyticum concentration and contact time were observed. Scanning electron microscopy examination showed masses of U. urealyticum attached to the head and middle piece of some of deformed spermatozoa. It is suggested that U.urealyticum is involved in sperm changes leading to male infertility, particularly when there is heavy U. urealyticum colonization or specific infections with this microorganism.
Hum Reprod. 1998 Oct;13(1O):2756-61.
Objective evaluation of the morphology of human epididymal sperm heads
Soler C, Perez-Sanchez F, Schulze H, Bergmann M, Oberpenning F, Yeung C, Cooper TG.
Department of Animal Biology, University of Valencia, Spain; Institute of Reproductive Medicine of the University, Munster, Germany.
Spermatozoa were obtained from nine epididymal regions of six epididymides taken from five men undergoing castration for prostatic carcinoma (53-76 years) and from one man with testicular cancer (38 years). Spermatozoa were obtained by mincing tissue in phosphate-buffered saline, making air dried smears and staining with Hemacolor. The percentage of sperm heads categorised subjectively as normal (of uniform shape) or otherwise was calculated for each region. This confirmed that grossly swollen sperm heads (previously shown to be artefacts) were only present in proximal regions of the duct. A computer-aided sperm morphology analyser (Sperm Class Analyzer(R)) was used to provide objective measurements of sperm head area, perimeter, length and width of the uniform sperm heads and revealed that there was a gradual and statistically significant decline in sperm head size upon maturation, as occurs in other species. There was no significant difference between the morphometric parameters of spermatozoa obtained from the distal cauda epididymis and those obtained from the ejaculates of young normozoospermic patients.
Int J Androl. 2000 Apr;23(2):77-84.
Human Sperm in The Third Dimension
Evaluation of automated system Sperm Class Analyzer (SCA) for semen analysis
Carlos Aulesa, M. Cabrera, R. Alonso, M. Benítez y M. Martínez
Unidad de Seminología, Laboratorios Clínicos, Ciutat Sanitària Vall d’Hebron, Barcelona, España
Objective
Evaluation of the efficiency of the New Sperm Class Analyzer® (SCA v.3.2.0) in sperm automated analysis for the calculation of the motility, concentration and morphology parameters using the latest CASA (Computer-Assisted Sperm Analysis) technology in software for spermiogram analysis.
Materials and methods
Analysis of 150 semen samples from the clinical areas of Fertility and Urology, 42 samples were analyzed with the SCA concentration module and 65 samples were analyzed with the SCA motility module using a disposable 10 micron deep Leja Chamber in both. At the same time the samples were analyzed by the manual method using the Neubauer Improved chamber for counting and slides with 22×22mm cover slip heated to 37°C with an average of 200 spermatozoa for motility, in order to test the precision, linearity and accuracy of SCA system compared with the manual method. The morphology analysis was evaluated using 50 pre-stained slides and evaluating 200 cells both manually by an ESHRE qualified technician and the SCA method and then calculating the correlation coefficient.
Results
The counting with SCA system compared with the manual method has shown a within-day imprecision of 10.13% with a range between 3.2% and 17.1% depending on the count level; correlation with the manual method was r=0.98 (P=0.001). The linearity of the SCA was shown to be linear between 0.5 million and 190 million sperm/ml. The motility module showed a higher within-day imprecision in counting WHO ¿type b¿ and ¿type c¿ spermatozoa (21.81%) than the ¿type a¿ and ¿type d¿, with an average value of 10.32%, also depending on the count rate. The reliability of the motility parameter was evaluated up to a of 120 million sperm/ml. The morphology module, with a customised configuration of parameters, had a significant positive correlation (r=0.899; P=0.0001) compared with the manual method.
Conclusions
The comparison of the concentration and motility of spermatozoa between the manual method and the SCA modules using 10 microns deep Leja chambers was positively evaluated. A series of premises are established for using the automated morphology performed on the Sperm Class Analyzer. The clinical usefulness of some of the new kinetic and morphometric parameters of sperm provided by the semen analyser is also established.
Laboratorio Clinico. 2009;02:8-16.
Morphometric dimensions of the human sperm head depend on the staining method used
L. Maree, S.S. du Plessis, R. Menkveld and G. van der Horst
Department of Medical Biosciences, University of the Western Cape, Private Bag X17, Bellville 7530, South Africa. Division of Medical Physiology, Department of Biomedical Sciences, Stellenbosch University, PO Box 19063, Tygerberg 7505, South Africa. Andrology Laboratory, Department of Obstetrics and Gynaecology, Tygerberg Academic Hospital and Stellenbosch University, Tygerberg 7505, South Africa
BACKGROUND: Assessment of sperm morphology (including morphometry) is extensively used to determine one of the qualities of a semen sample and depends on the differential staining of spermatozoa. A staining technique should cause as little change to sperm dimensions and form as possible in order to reliably evaluate the morphometric features of the sperm. Various staining techniques have been employed, but only a few have been recommended by the World Health Organization and are amenable to automated sperm morphometry analysis. Our study was aimed at comparing the effect of three staining techniques [Papanicolaou (PAP), Rapidiff® (RD) and SpermBlue® (SB)] on human sperm head dimensions and to compare these with the head dimensions in fresh semen.
METHODS: Smears made from human semen samples (n = 24) were stained according to the three staining techniques and sperm head morphometry was assessed with the Sperm Class Analyzer. Head dimensions of fresh spermatozoa were measured with a digital calliper on a computer screen. The minimum number of spermatozoa to be analyzed to represent the sperm population and the degree of inter-laboratory variation were determined. Electron micrographs from the same semen samples were used to determine the actual acrosome coverage of the spermatozoa in the semen (n = 7) in order to verify the results of the automatic analyses.
RESULTS: The osmolality of human semen differs from that of the RD and PAP fixatives and stains, but is more similar to the SB fixative and stain. At least 100 spermatozoa should be analyzed to include a representative sample of the sperm population. RD caused sperm heads to swell, PAP caused them to shrink and SB had no significant effect on sperm head dimensions when compared with spermatozoa in fresh semen. Very little inter-laboratory variations were found. The percentage acrosome coverage was significantly different between the three staining techniques, as well as between the RD and PAP stains and the manual measurements obtained using the electron micrographs.
CONCLUSIONS: Different staining techniques change the morphometric dimensions of the human sperm head, probably due to the fact that either the fixatives or stains are not iso-osmotic in relation to human semen. Since these changes in sperm head dimensions are not uniform, care should be taken when selecting a staining technique. Ideally, stained spermatozoa should have dimensions as close to spermatozoa in fresh semen as possible, as was found with the SB staining method, resulting in accurate evaluations of sperm head morphometry.
Human Reproduction 2010 25(6):1369-1382; doi:10.1093/humrep/deq075
SpermBlue®: A new universal stain for human and animal sperm which is also amenable to automated sperm morphology analysis
G van der Horst; L. Maree
Department of Medical Biosciences, University of the Western Cape, Private Bag X17, Bellville, South Africa
Our study was aimed at exploring a simple procedure to stain differentially the acrosome, head, midpiece, and flagellum of human and animal sperm. A further prerequisite was that sperm morphology of the stained samples could be analyzed using automated sperm morphology analysis (ASMA). We developed a new staining process using SpermBlue® fixative and SpermBlue® stain, which are iso-osmotic in relation to semen. The entire fixation and staining processes requires only 25 min. Three main steps are required. First, a routine sperm smear is made by either using semen or sperm in a diluting medium. The smear is allowed to air dry at room temperature. Second, the smear is fixed for 10 min by either placing the slide with the dried smear in a staining tray containing SpermBlue® fixative or by adding 1 ml SpermBlue® fixative to the slide. Third, the fixed smear is stained for 15 min by either immersing the slide in a staining tray containing SpermBlue® stain or adding four drops of SpermBlue® stain to the fixed smear. The stained slide is dipped gently in distilled water followed by air drying and mounting in DPX® or an equivalent medium. The method is simple and suitable for field conditions. Sperm of human, three monkey species, horse, boar, bull, ram, mouse, rat, domestic chicken, fish, and invertebrate species were stained successfully using the SpermBlue® staining process. SpermBlue® stains human and animal sperm different hues or intensities of blue. It is possible to distinguish clearly the acrosome, sperm head, midpiece, principal piece of the tail, and even the short end piece. The Sperm Class Analyzer® ASMA system was used successfully to quantify sperm head and midpiece measurements automatically at either 600 times or 1000 times magnification for most of the species studied.
Biotechnic and Histochemistry, Volume 84, Issue 6 December 2009 , pages 299 - 308
Clinical Implementation of the Halosperm® Test Kit in Combination with SCA®
Kellie Williams, PhD and J. Kevin Thibodeaux, PhD, HCLD
Tulsa Fertility Center, 115 East 15th St., Tulsa OK 74119
Fertility Magazine, Volume 12, pages 66-68
Seminal diagnostic
A. Sebastianelli, L. Caponecchia
Unit of Andrology and Physiopathology of Reproduction, Centre for Sterility in Couples and Cryopreservation
of Gametes, "S. Maria Goretti" Hospital, ASL of Latina, Italy
Journal of Andrological Sciences 2008;15(suppl 1):18-23
The in vitro effect of benzo[a]pyrene on human sperm hyperactivation and acrosome reaction
Dyutiman Mukhopadhyay, M.S., Parag Nandi, M.S., Alex C. Varghese, Ph.D., Rohit Gutgutia, M.D., Samir Banerjee, Ph.D., Asok K. Bhattacharyya, Ph.D., D.Sc.
Department of Zoology, University of Calcutta; Institute of Reproductive Health and Toxicology; Department of Environmental Science, University of Calcutta; IVF Division, Advanced Medicare and Research Institue Ltd.; Department of Biochemistry, University of Calcutta, Calbutta, India
Objective
To evaluate the in vitro effect of benzo[a]pyrene on sperm hyperactivation and acrosome status in normozoospermic semen samples of nonsmokers analyzed by computer-assisted semen analysis (CASA).
Design
Experimental in vitro study.
Setting
Andrology laboratory.
Patient(s)
Thirteen proven fertile, normozoospermic, and nonsmoking men.
Intervention(s)
Spermatozoa were washed free of seminal plasma and were treated with different concentrations of benzo[a]pyrene and compared with controls treated with medium alone. The benzo[a]pyrene concentrations were: 100, 50, 25, and 12.5 µg/mL.
Main Outcome Measure(s)
Effect of varying concentrations of benzo[a]pyrene on sperm hyperactivation and acrosomal reaction.
Result(s)
A statistically significant increase in sperm hyperactivation was observed in presence of benzo[a]pyrene at concentrations of =50 µg/mL. The result of the acrosome halo test showed that concentrations of benzo[a]pyrene =25 µg/mL statistically significantly decreased the percentage of halo formation, indicating an inappropriate (false) acrosome reaction.
Conclusion(s)
Benzo[a]pyrene statistically significantly affected sperm functional competence as evidenced by increased hyperactivation as well as premature acrosomal reaction.
Presented at the 64th annual meeting of American Society of Reproductive Medicine (ASRM), November 8-12, 2008, San Francisco, California.
(Gutgutia R, et al. In vitro analysis of the effect of benzo[a]pyrene on sperm hyperactivation [abstract]. Fertil Steril 2008;90:S185.)
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