Biomedical subjects

P N Zarmakoupis

Publications and source records attributed to P N Zarmakoupis.

6 recordsLinked to original sources

PubMed1995

Inhibition of human endometrial stromal cell proliferation by interleukin 6.

We investigated the ability of interleukin 6 (IL-6) to modulate human endometrial stromal cell growth in vitro. Stromal cell proliferation in response to treatment with varying concentrations of IL-6 was determined. Endometrial tissue was obtained from 10 normally cycling women during the secretory phase of their menstrual cycle. Treatment with IL-6 resulted in a dose- and cell-density-dependent inhibition of endometrial stromal cell proliferation in vitro. The maximal inhibition was observed with 200 pg/ml of IL-6 and at a concentration of 10(5) cells/well. During in-vitro culture, stromal cells produced low amounts of IL-6 and demonstrated the presence of IL-6 receptor. These data demonstrate that IL-6 acts as a growth-regulatory signal for human endometrial stromal cells. We postulate that IL-6 may contribute to the maintenance of homeostasis in normal endometrium and that perturbation of IL-6 mediated responses may play a role in disorders of the endometrium such as endometrial cancer and endometriosis.

Cell Count↗

PubMed1995

Dysregulation of interleukin-6 responses in ectopic endometrial stromal cells: correlation with decreased soluble receptor levels in peritoneal fluid of women with endometriosis.

Endometriosis (EM) is characterized by the aberrant growth of endometrial cells at sites outside the uterus. We showed previously that peritoneal leukocyte interleukin-6 (IL-6) production is altered in women with EM relative to that in normal control women. Because studies suggest that IL-6 may be growth regulatory for endometrial cells, we examined IL-6 and IL-6 soluble receptor (IL-6sR) in the peritoneal fluid of 40 women. In addition, the growth responsiveness of ectopic endometrial stromal cells to IL-6 was evaluated. The severity of EM correlated with increased levels of IL-6 accompanied by decreased IL-6sR in peritoneal fluid (controls, 1.0 +/- 0.1 and 525.4 +/- 53; stage I-II EM, 1.4 +/- 0.2 and 274.6 +/- 26; stage III-IV EM, 19.3 +/- 4.6 and 319.4 +/- 26; adhesions, 1.9 +/- 0.4 and 324.7 +/- 26 pmol/L IL-6 and IL-6sR, respectively). Additional studies revealed that unstimulated endometrial stromal cells from ectopic implants secreted this cytokine in vitro. Furthermore, these cells were resistant to growth inhibition induced by exposure to additional IL-6; this response correlated with weak expression of IL-6 receptor. Taken together, these findings lend further support to the hypothesis that dysregulation of IL-6 responses plays a role in the pathophysiology of EM.

Adolescent↗

PubMed1995

Epididymal spermatozoa: recovery and subsequent improvements of mouse epididymal spermatozoa via the SpermPrep filtration method.

This study was designed to determine the effects of Sephadex filtration (SpermPrep method) on the separation of viable motile, morphologically normal mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3 hr incubation period. Spermatozoa were harvested from the caudae epididymides (5 animals per run or replication; n = 10) following bilateral testicular excision and incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15 min. The specimen was then split into 2 aliquots, with Alquot 1 as the control and Aliquot 2 used for filtration. SpermPrep I column was employed according to the manufacturer's specifications (ZBL, Inc., Lexington, KY, USA) using TYB. During filtration (10 min), different fractions were obtained: first 5 min (Sample 1) and second 5 min (Sample 2). The filtered fractions were evaluated and incubated at 37 degrees C and assessed for percentage and grade of motility (0-4) every 30 min for 3 hr. Filtration resulted in a significant improvement in percentage and grade of motility (91.5% and 3.0 vs. 76.5% and 2.5, respectively). The results point out very clearly that the filtration via the SpermPrep method improved the percentage and grade of motility (p < 0.05) but not the percentage normal morphology of the spermatozoa. Also the SpermPrep I enabled the recovery of 45% (8.3 x 10(6) spermatozoa) of the total spermatozoa processed in the control aliquot (18.4 x 10(6) spermatozoa) which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3 hr showed greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs. 39% and 0.82, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

A method of short-term cryostorage and selection of viable sperm for use in the various assisted reproductive techniques.

The objective of this study was to determine if spermatozoa, following short-term cryostorage at 5 degrees C in Test-Yolk buffer (TYB; ZBL, Inc., Lexington, KY, USA), could be recovered and improved via the SpermPrep filtration method and to assess the possibly enhanced fertilizing capacity of the selected spermatozoa. Semen specimens from 20 men were collected, evaluated, diluted 1:1 (v/v) with TYB, divided into aliquots and cooled to 5 degrees C for 24 and 48 hr. Semen samples were assessed for volume, sperm count, percentage and grade of motility, percentage of morphologically normal spermatozoa and outcome of the sperm penetration assay (SPA). After storage, aliquots were rewarmed at 37 degrees C, centrifuged, and the pellet was resuspended in 1.0 ml of SpermPrep media (ZBL, Inc.). Following 15 min of incubation, the rewarmed spermatozoa were filtered via the SpermPrep I filtration column (ZBL, Inc.) and assessed accordingly. The results obtained in this study indicate that the short-term cryostorage procedure yielded spermatozoa of adequate qualitative characteristics when compared to the fresh spermatozoa. Furthermore, filtration of rewarmed specimens yielded spermatozoa of significantly higher qualitative characteristics and superior fertilizing capacity following a short-term cryostorage period in TYB when compared to fresh and rewarmed spermatozoa (p < 0.05). This method of short-term cryostorage in TYB and selection of superior spermatozoa via the SpermPrep filtration method could further enhance the fertilizing ability of patients who produce spermatozoa characterized by deficient capacitation, acrosome reaction and subsequent fertilization.

Cryopreservation↗

PubMed1994

Improvements and short-term viability of mouse epididymal spermatozoa recovered through the Spermprep filtration method.

This study was designed to determine the effects of Sephadex filtration (SpermprepI method) on the separation of motile, morphologically normal, mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3-h incubation period. Spermatozoa were harvested from the caudae epididymie (5 animals per run or replication; n=10) following bilateral testicular excision, after which they were incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15-min. The specimens were then split into 2 1-ml aliquots, with Aliquot 1 as the control and Aliquot 2 as the filtered sample. The SpermprepI column was employed according to the manufacturer's specifications using TYB. During filtration (10-min), 2 different fractions were obtained: first 5-min (Sample 1) and second 5-min (Sample 2). The 2 fractions were evaluated and incubated at 37 degrees C and assessed for percentage of motility and grade of motility (0 to 4) every 30-min for 3-h. Filtration resulted in a significant improvement in the percentage and grade of motility (91.5% and 3.0 vs 76.5% and 2.5, respectively). The results indicate that filtration with the SpermprepI method improved the percentage and grade of motility (P<0.05) but not the percentage of normal morphology of the spermatozoa. In addition, the SpermprepI method enabled the recovery of 45% (8.3x10(6) spermatozoa recovered) of the total number of spermatozoa processed in the control aliquot (18.4x10(6) spermatozoa), which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3-h showed a greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs 39% and 0.82, respectively. The SpermprepI filtration method selected a higher proportion of quality spermatozoa, which also displayed significant long-term motility (longevity) during in vitro incubation.

Journal Article↗

PubMed1994

Differences in seminal parameters in specimens collected via intercourse and incomplete intercourse (coitus interruptus)

Whatever the mechanism of such improvements may be, the results in this study point out that coitus interruptus in the human may not be the method of choice for collection of semen specimens, especially in patients with spermatogenic dysfunctions such as hypospermia, oligospermia, and asthenospermia. It should also be noted at this point that, for whatever purpose (semen analysis or artificial insemination by husband), the collected specimen should as closely as possible resemble the ejaculate delivered during intercourse. The complete coitus method, as applied in this study, showed that completion of the ejaculatory process during intercourse as compared with the coitus interruptus method, may assist in the improvement of the collected specimen and should closely resemble the ejaculate obtained during intercourse without the use of Silastic condoms. Furthermore, on the basis of the results generated in this study, the complete coitus method should always be the method of choice for male infertility patients with ejaculatory and spermatogenic dysfunctions as well as for scientists and clinicians who deal in the field of infertility diagnosis and treatment.

Coitus↗