Comparison of three staining methods for the morphological evaluation of human spermatozoa

Objective: To compare different staining methods to evaluate human sperm morphology. Design: Prospective study. Setting: Patients at the Departments of Dermatology and Urology, University of Jena, Germany. Patient(s): A total of 94 randomly collected patients attending the andrological outpatient clinics of the Departments of Dermatology and Urology, University of Jena, Germany. Intervention(s): None. Main Outcome Measure(s): Statistical comparison of resultant standard morphological parameters (mean percentages) after staining according to Papanicolaou and Shorr methods and with Testsimplets® prestained slides. Result(s): All morphological parameters investigated (percent normal morphology, percent head, midpiece, and flagellar abnormalities) correlated statistically significantly positively, however with markedly lower correlation coefficients for the Testsimplets®results. As compared with the mean Papanicolaou (4.78% ± 2.54%) and Shorr staining (4.75% ± 2.64%) results, a statistically significantly lower percentage of morphologically normal spermatozoa was determined after using the Testsimplets® slides (3.89% ± 2.53%). In general, the mean values of all parameters differed for all comparisons with the Testsimplets® slides and especially for the percentage of flagellar defects but not between the Papanicolaou and the Shorr staining results. Conclusion(s): The results show an extensive agreement between the Papanicolaou- and Shorr-stained smears, whereas Testsimplets® staining exhibited statistically significant deviations. Because the correct evaluation of sperm morphology is of essence within the scope of assisted reproduction and in andrological diagnostics, the use of rapid staining methods cannot be recommended. © 2008 American Society for Reproductive Medicine.Articl

Typha capensis (Rohrb.)N.E.Br. (bulrush) extract scavenges free radicals, inhibits collagenase activity and affects human sperm motility and mitochondrial membrane potential in vitro: A pilot study

The biodiversity in South Africa provides more than 30000 higher plants, of which more than 3000 are used by traditional healers to treat diseases. Typha capensis (bulrush) is one of the medicinal plants used in South Africa to treat male fertility problems. Considering that South African traditional healers have been recognised by Law and the health benefits of T. capensis have not been scientifically investigated yet, this study aimed at investigating the in vitro effects of aqueous extracts from this plant on male reproductive functions. Both leaves and rhizomes of T. capensis were dried, infused with distilled water and freeze-dried. Motile sperm from 50 men were isolated by swim-up and incubated with 1μgml -1 aqueous extract of Typha rhizome for 1h at 37°C. Vitality, motility, sperm production of reactive oxygen species and mitochondrial membrane potential were analysed in the test sample, a control and in the pellet from the swim-up. Results showed that the rhizome extract had significant (P<0.0001) negative effects on all parameters. The extracts from the leaves and rhizomes revealed dose-dependent inhibitory activity for collagenase and free radical formation. No inhibitory activity for elastase was found. The inhibitory activity for collagenase might indicate possible anti-cancer effects. © 2011 Blackwell Verlag GmbH

Optimized protocol for the biocompatibility testing of compression stockings and similar products with close skin contact in vitro

Biocompatibility of six different compression stockings and cytotoxic effects were determined using HaCaT keratinocytes, L929 mouse fibroblasts, primary adult and juvenile keratinocytes Cells were quantified using a luminometric ATP assay and the photometric BCA test. Cytotoxic effects were determined by LDH release. An area-based extraction ratio of 1.25 cm2:mL could be shown to be superior to the weight-based extraction of test material. Extraction medium should be an acidic sweat solution as this helps to recreate in vivo conditions. Monolayer cultures of HaCaT keratinocytes or L929 mouse fibroblasts should be used for testing. Primary adult keratinocytes or primary juvenile keratinocytes can also be used. For the latter, testing under DMEM with FCS is recommended to achieve comparable results. It was found that the compression stockings tested exhibited no negative influence on cell viability in vitro and no direct cytotoxic effects measured as release of LDH. Henc e, good biocompatibility could be asserted