Stability of αB-crystallin gene expression in canine mammary gland neoplasms. Should it be considered as circulating tumor cell genetic marker?

αB-crystallin is a member of a small family of thermal shock proteins that protects cells from stress. Because of lack of its expression in peripheral blood leukocytes, it was proposed as a molecular marker of circulating tumor cells in canine mammary gland tumors. The aim of the present study was to determine if αB-crystallin shows stability of expression, what is the requirement for this type of marker. It was also assessed whether there is co-expression of αB-crystallin with the basal marker, cytokeratin 17. For this purpose, samples of various types of canine mammary gland tumors of epithelial origin, were selected. Using RT-qPCR, we have found αB-crystallin and cytokeratin 17 co-expression in benign and malignant canine mammary gland tumors. It has been demonstrated that the expression of αB-crystallin in tested neoplastic samples is not stable in comparison to the control group. Furthermore αB-crystallin overor down- expression was associated witch the same cytokeratin 17 pattern. αB-crystallin can be a marker of circulating tumor cells in the bloodstream, but for cancers in which basal marker expression occurs and thus not universal for all cancers originating from the mammary gland tissue.

Serum progesterone concentrations on the day of oocyte retrieval above 9.23 ng/ml may predict ovarian hyperstimulation syndrome risk in in vitro fertilized patients.

The aim of our study was to evaluate serum progesterone levels on the day of oocyte retrieval as a promising biomarker inorder to evaluate the risk of ovarian hyperstimulation syndrome in a group with controlled ovarian hyperstimulation protocols using either gonadotropin antagonists or agonists (GnRH), compare with a natural cycle control group. Patients were divided into 3 groups (148 patients in total): control group in the natural cycle, patients treated with GnRH agonist and patients treated with GnRH antagonist. When we compared both controlled ovarian hyperstimulation (COH) protocol groups with the control group, we found statistically higher levels of progesterone in patients after COH (control versus long protocol group: 1.43 ± 1.28 ng/ml versus 8.95 ± 5.95 ng/ml; P < 0.001; control versus GnRH antagonist group: 1.43 ± 1.28 ng/ml versus 7.18 ± 5.13 ng/ml; P < 0.001). According to receiver operating characteristic (ROC) analysis, the level of serum progesterone on the day of oocyte retrieval, above which the risk of ovarian hyperstimulation syndrome (OHSS) is associated with a more than fourfold higher risk (OR 4.24; 95% CI 2.6 - 6.9) was found to be 9.23 ng/ml, with AUC: 0.896, P = 0.026 (95% CI 0.845 - 0.947). Progesterone level on the day of oocyte retrieval may be used as an additional sensitivity marker in treatment of early forms as well by freezing of embryos in prevention of late forms of OHSS.

Stress urinary incontinent women, the influence of age and hormonal status on estrogen receptor alpha and beta gene expression and protein immunoexpression in paraurethral tissues.

The underlying cause of stress urinary incontinence (SUI) is an anatomical abnormality associated with paraurethral connective tissue dysfunction. The question as to whether estrogens affect the quality of that tissue remains unexplained. Samples of paraurethral connective tissue from 81 women were examined (the SUI's n = 49; the control's n = 32). In both groups, the patients were subdivided into pre- and postmenopausals. Primary study outcome was comparison of the estrogen receptor alpha (ERα) and the estrogen receptor beta (ERß) gene and protein in paraurethral tissue between SUI and control group. Secondary study outcome was comparison of these receptors according to hormonal status of the patients and their age. In both examined groups, we found both ER proteins. The ERα gene expression was detected in-19/32 (SUI) samples and in 24/31 (control), and ERß gene expression 31/32 and 30/31 samples, respectively. The SUI's had significantly lower ERa gene expression premenopausally than the control's. The analysis found considerably lower ERß and reduced ERα gene expression in postmenopausals, approaches the significance level. There was also significant decrease in both receptors' genes expression in post-53 women, compared to younger patients. Spearman's correlation test revealed a statistically significant decrease in ERß gene with age. Both estrogen receptors are found in women's paraurethral tissue, so this tissue is an estrogen target. No correlation between ERß gene expression and immunoexpression and SUI was found. The ERα gene seems to play a key role in SUI in the premenopausal period, but ERß gene expression in the paraurethral connective tissue decreases with age.

High levels of soluble vascular endothelial growth factor receptor 1/sFlt1 and low levels of vascular endothelial growth factor in follicular fluid on the day of oocyte retrieval correlate with ovarian hyperstimulation syndrom regardless of the stimulation protocol.

The aim of the study was to assess the predictive value of vascular endothelial growth factor (VEGF), its soluble receptor - sVEGF-R1/sFlt1 and endocrine gland-derived vascular endothelial growth factor (EG-VEGF) concentrations in serum and follicular fluid (FF) for ovarian hyperstimulation syndrome (OHSS) in women undergoing controlled ovarian hyperstimulation (COH) protocols. Patients have been divided into 3 groups: control group on natural cycle, patients stimulated with GnRH agonist and patients stimulated with GnRH antagonist. The FF and serum concentrations of VEGF, EG-VEGF, sVEGF R1 and the expression of VEGF and EG-VEGF mRNA in GC in small and large follicles collected from patients were investigated. When we compared all patients in a trial, OHSS occurrence was correlated with higher level of sVEGF R1 and a lower level of VEGF in a follicular fluid from large follicles in a day of oocyte retrieval. The VEGF/sVEGF-R1 ratio for patients in COH groups, above which the risk of developing OHSS is very low (OR 0.1 (95% CI 0.01 - 0.29, P = 0.0006) was found to be 0.281 pg/ml, with AUC - 0.738, P = 0.042, (95% CI 0.656 - 0.82). High levels of sVEGF-R1 and low level of VEGF in FF on the day of oocyte retrieval correlate with OHSS regardless of the stimulation protocol.

Pain assessment during outpatient hysteroscopy using room temperature versus warm normal saline solution as a distention medium - a prospective randomized study.

OBJECTIVE: To assess the efficacy of warm normal saline distention solution versus a standard, room-temperature normal saline as distention medium for pain relief during outpatient hysteroscopy. MATERIALS AND METHODS: A prospective randomized case-placebo controlled study was conducted in tertiary care centre - Central Clinical Hospital of Ministry of Interior and Administration. Study group consisted of 100 women referred for outpatient hysteroscopy between January 2015 and July 2015. Every patient, who was referred for an office hysteroscopy, was offered to participate in the study to receive a sterile, 0.9% normal saline warmed up to 36°C as distention medium. Control group were women receiving sterile, room temperature of 25°C, 0.9% normal saline solution as a distention medium. No pre-medication nor analgesia were used. A visual analogue scale (VAS) was used for one-dimensional pain assessment. Women were asked to mark a VAS score before, during, and five and 15 minutes following the procedure. RESULTS: Median VAS scores during and directly after the anaesthesia-free hysteroscopy were no different between two groups. (p = 0.554 and p = 0.121, respectively). There were also no differences in the procedure time between groups (p = 0.845). CONCLUSIONS: Warm normal saline distention solution does not reduce the pain during and at the end of the outpatient hysteroscopy. The effect does not depend on the age of women, menopausal status, parity or type of outpatient hysteroscopy (operative or diagnostic).

Galanin regulates blood glucose level in the zebrafish: a morphological and functional study.

The present study has demonstrated the galaninergic innervation of the endocrine pancreas including sources of the galaninergic nerve fibers, and the influence of galanin receptor agonists on blood glucose level in the zebrafish. For the first time, a very abundant galaninergic innervation of the endocrine pancreas during development is shown, from the second day post-fertilization to adulthood. The fibers originated from ganglia consisting of galanin-IR, non-adrenergic (non-sensory) neurons located rostrally to the pancreatic tissue. The ganglia were found on the dorsal side of the initial part of the anterior intestinal segment, close to the intestinal branch of the vagus nerve. The galanin-IR neurons did not show immunoreactivity for applied antibodies against tyrosine hydroxylase, choline acetyltransferase, and vesicular acetylcholine transporter. Intraperitoneal injections of galanin analog NAX 5055 resulted in a statistically significant increase in the blood glucose level. Injections of another galanin receptor agonist, galnon, also caused a rise in blood glucose level; however, it was not statistically significant. The present findings suggest that, like in mammals, in the zebrafish galanin is involved in the regulation of blood glucose level. However, further studies are needed to elucidate the exact mechanism of the galanin action.

Zebrafish: an animal model for research in veterinary medicine.

The zebrafish (Danio rerio) has become known as an excellent model organism for studies of vertebrate biology, vertebrate genetics, embryonal development, diseases and drug screening. Nevertheless, there is still lack of detailed reports about usage of the zebrafish as a model in veterinary medicine. Comparing to other vertebrates, they can lay hundreds of eggs at weekly intervals, externally fertilized zebrafish embryos are accessible to observation and manipulation at all stages of their development, which makes possible to simplify the research techniques such as fate mapping, fluorescent tracer time-lapse lineage analysis and single cell transplantation. Although zebrafish are only 2.5 cm long, they are easy to maintain. Intraperitoneal and intracerebroventricular injections, blood sampling and measurement of food intake are possible to be carry out in adult zebrafish. Danio rerio is a useful animal model for neurobiology, developmental biology, drug research, virology, microbiology and genetics. A lot of diseases, for which the zebrafish is a perfect model organism, affect aquatic animals. For a part of them, like those caused by Mycobacterium marinum or Pseudoloma neutrophila, Danio rerio is a natural host, but the zebrafish is also susceptible to the most of fish diseases including Itch, Spring viraemia of carp and Infectious spleen and kidney necrosis. The zebrafish is commonly used in research of bacterial virulence. The zebrafish embryo allows for rapid, non-invasive and real time analysis of bacterial infections in a vertebrate host. Plenty of common pathogens can be examined using zebrafish model: Streptococcus iniae, Vibrio anguillarum or Listeria monocytogenes. The steps are taken to use the zebrafish also in fungal research, especially that dealing with Candida albicans and Cryptococcus neoformans. Although, the zebrafish is used commonly as an animal model to study diseases caused by external agents, it is also useful in studies of metabolic disorders including fatty liver disease and diabetes. The zebrafish is also a valuable tool as a model in behavioral studies connected with feeding, predator evasion, habituation and memory or lateralized control of behavior. The aim of the present article is to familiarize the reader with the possibilities of Danio rerio as an experimental model for veterinary medicine.

Accuracy and diagnostic value of outpatient hysteroscopy for malign and benign disease.

BACKGROUND: The aim of the study was to evaluate accuracy of the outpatient hysteroscopy. MATERIALS AND METHODS: This was a retrospective cohort study of 494 women who underwent outpatient hysteroscopy after administration ofnon-steroidal anti-inflammatory agents with the 3.2 mm hysteroscope. Normal saline solution was used as the distension medium. All women were discharged in good general condition afterwards. RESULTS: In cases of abnormal uterine bleeding, there was no apparent pathology found in 112 cases (83.6%). Detection rate (DR) of endometrial polyps was 88.7% with false positive rate (FPR) of 4.6%. Positive predictive value (PPV) was 82.7% with negative predictive value (NPV) of 93.1%. Detection rate (DR) of the submucosal fibroids was 57.7%. Positive predictive value (PPV) was 57.7% with negative predictive value (NPV) of 95.0%. Endometrial cancer was confirmed in ten cases (2.0%), being suspected in eight cases during the procedure. DR in case of the endometrial cancer was 80.0% with FPR of 0.4%. PPV was 66.7% with NPV) of 99.6%. CONCLUSIONS: Outpatient hysteroscopy seems to be an effective and accurate diagnostic tool.

High levels of women's satisfaction and compliance with transdermal contraception: results from a European multinational, 6-month study.

OBJECTIVE: To investigate compliance, satisfaction, and preference in women using a transdermal contraceptive patch. METHODS: Women (18-46 years) from eight European countries used contraceptive patches (norelgestromin 6 mg, ethinylestradiol 600 µg) for six, 4-week treatment cycles. Compliance, satisfaction, and preference were assessed after 3 and 6 cycles and study completion using self-report methods. RESULTS: Of the 778 participants, 36.8% (n = 287) used no contraception at baseline. The most common methods were oral contraceptives (67.9%, n = 334) and barrier methods (21.5%, n = 106). Of oral contraception users, 63.5% (n = 212) were satisfied or very satisfied with their previous method, but compliance was poor with 77.8% (n = 260) reporting missed doses. After 3 and 6 cycles, >80% of all included women were satisfied or very satisfied with the patch. At study completion, most participants (73.7%) reported a preference for the patch compared to their previous method. Of 4107 cycles, 3718 (90.5%) were completed with perfect compliance. Two pregnancies occurred during this study, representing a Pearl Index of 0.63. No new safety issues were identified and the patch was well tolerated. CONCLUSION: Women were highly satisfied with transdermal contraception and preferred this form of family planning over their previous method. Transdermal contraception represents a valuable addition to contraceptive options with potential to offer high compliance and efficacy.

Leptin gene expression in subcutaneous adipose tissue in girls before and during puberty.

OBJECTIVES: Leptin functions as a neuroendocrine hormone and it is related to the onset of puberty in animal models. Its role in normal human sexual maturation is still incompletely defined. The aim of the study was to assess the relationships between leptin mRNA (gene) expression, thickness of subcutaneous fat tissue and the serum concentration of leptin in girls before and during puberty. STUDY DESIGN: Twenty-nine lean girls were studied (mean age 10.8+/-1.9 years). The subjects were divided into two groups according to pubertal status. The first group consisted of 14 prepubertal girls and second group of 15 girls who were in puberty. Body height, weight, arm circumference, skin fold thickness at abdominal, triceps and subscapular sites were measured. Serum leptin was assessed by RIA method. Leptin mRNA was measured in subcutaneous abdominal adipose tissue by semi-quantitative assays based on reverse transcription (RT) of the mRNA and polymerase chain reaction (PCR) amplification of the cDNA. RESULTS: Girls in pubertal stages had higher serum leptin concentration than prepubertal girls. The mean values of leptin mRNA level in subcutaneous abdominal adipose tissue were not statistically different between groups. There was also no difference between the thickness of skin folds in investigated girls. A positive correlation between leptin mRNA expression and skin fold thickness, BMI and arm circumference as well as between the leptin concentration and skin fold thickness, BMI and arm circumference were observed. CONCLUSIONS: The level of leptin gene expression and serum leptin concentrations depend on the amount of fat tissue. We can propose that initiation of pubertal events does not result from increased of leptin mRNA expression in subcutaneous abdominal fat cells or from its increased concentration in blood.

Contraceptive efficacy and safety of DMPA-SC.

DMPA-SC 104 mg/0.65 mL is a new, low-dose subcutaneous (SC) formulation of Depo-Provera contraceptive injection (150 mg/mL medroxyprogesterone acetate injectable suspension) that provides efficacy, safety and immediacy of onset equivalent to Depo-Provera intramuscular (IM) injection. Two large, open-label, Phase 3 studies assessed the 1-year contraceptive efficacy, safety and patient satisfaction with DMPA-SC administered every 3 months (12-13 weeks). Zero pregnancies were reported in both studies, which included a total of 16,023 woman-cycles of exposure to DMPA-SC and substantial numbers of overweight or obese women. DMPA-SC was well-tolerated and adverse events were similar to those reported previously with Depo-Provera IM. Thus, DMPA-SC offers women a new, highly effective and convenient long-acting contraceptive option.

Selective alterations in insulin receptor substrates-1, -2 and -4 in theca but not granulosa cells from polycystic ovaries.

The elevated insulin concentrations that occur in many women with polycystic ovary syndrome (PCOS) can contribute significantly to ovarian hyperandrogenism. The objective of the present study was to compare the content of proximal insulin signalling molecules in theca and granulosa cells between polycystic ovaries and regular cycling controls. Individual follicles (3-7 mm) were obtained from 11 women with PCOS and 10 regularly cycling control women. The theca and granulosa cells were microdissected from each follicle. Total protein was extracted and signalling proteins were measured by western blot analysis. There was no difference in insulin receptor content between PCOS and controls in either theca or granulosa cells. Insulin receptor substrate (IRS)-1 and -2 were increased (P<0.05), but IRS-4 was decreased (P<0.03) in PCOS theca cells. There were no changes in IRS-1, -2 or -4 in granulosa cells. IRS-3 was undetectable in all samples. There were no changes in phosphatidyl inositol-3 kinase catalytic subunits p110alpha or p110beta in either theca or granulosa cells. These data demonstrate cell-specific alterations in IRS protein concentrations in theca cells from polycystic ovaries that are consistent with an exaggerated amplification of the insulin signal and which may play an important role in ovarian hyperandrogenism and thecal hyperplasia.

Leptin messenger ribonucleic acid (mRNA) content in the human placenta at term: relationship to levels of leptin in cord blood and placental weight.

During fetal life, placental tissue represents an additional source of leptin for the mother and conceptus. It has been suggested that feto-placental production of leptin may be involved in placental and fetal growth regulation. The aim of this study was to examine the correlation between leptin mRNA expression in the placenta and the concentrations of leptin in cord blood. A total of 30 healthy, pregnant women who gave birth to healthy neonates were included in the study. Maternal blood (obtained from the cubital vein) and umbilical cord blood were drawn immediately after birth. Serum leptin concentration was determined by enzyme-linked immunosorbent assay and serum insulin concentration was measured by radioimmunoassay. Leptin mRNA was measured in placental tissue by a reverse transcriptase-polymerase chain reaction. The estimated mean leptin mRNA expression in placenta was 4.65 +/- 1.83 pg mRNA/microg DNA. Leptin mRNA correlated with cord serum leptin concentrations (r = 0.3691, p = 0.045). Placental weight correlated with placental leptin mRNA (r = 0.3686, p = 0.045). The mean leptin concentration in cord serum at birth was slightly lower (3.1 +/- 1.9 ng/ml) than that found in maternal serum (3.9 +/- 1.2 ng/ml). A positive correlation was observed between cord and maternal serum leptin levels (r = 0.58, p = 0.001). The mean insulin concentration in maternal serum was not significantly higher than that in umbilical serum: 22.2 +/- 17.8 microIU/ml vs. 6.9 +/- 3.6 microIU/ml; r = 0.069, p = 0.71). Neither maternal nor umbilical insulin concentrations correlated with leptin concentration in cord or maternal peripheral serum.

Vertebral fractures: a hidden problem of osteoporosis.

Osteoporosis is a progressive skeletal disease, which in many cases remains silent and asymptomatic until a fracture occurs. Vertebral fractures are the earliest and most common osteoporotic fractures. The prevalence of vertebral fractures increases steadily with age, ranging between 20% for 50-year-old postmenopausal women to 64.5% for older women. The majority of vertebral fractures are not connected with severe trauma, and only one in three is diagnosed clinically. Usually vertebral fractures are associated with such clinical symptoms as back pain, posture change, loss of height, functional impairment, disability, and diminished quality of live. Women with the most severe vertebral fractures are the most likely to incur further fractures, with as much as 3.4 times the risk of hip fracture, and 12.6 times the risk of new vertebral fractures. Almost 20% of women will experience another fracture within 1 year after a vertebral fracture. Vertebral fractures are accompanied by increased mortality. The relative risk of death following vertebral fracture is almost 9 times higher. The most important purpose of osteoporosis management in postmenopausal women is prevention of the first vertebral fracture. Raloxifene (Evista) is the only SERM approved by the American FDA for the treatment and prevention of osteoporosis. It is the first compound with selective estrogen agonist activity in bone and in the cardiovascular system, but with estrogen antagonist activity or no activity in reproductive tissues and breast. Raloxifene reduces the risk of positive estrogen receptor breast cancer, decreases total cholesterol and LDL cholesterol, increases HDL cholesterol, does not increase the risk of endometrial cancer or cause bleeding and spotting. After 3 years of treatment Raloxifene reduces the risk of first vertebral fracture by 55%. The fracture risk within one year is reduced by as much as 68%. Continued observation has proved its sustained efficacy in the further reduction of fracture risk by 49% in the fourth year. Raloxifene treatment does not change the physiological structure of bone quality and does not cause fibrosis, osteomalacia or other toxic effects.

Luteinizing hormone receptor, steroidogenesis acute regulatory protein, and steroidogenic enzyme messenger ribonucleic acids are overexpressed in thecal and granulosa cells from polycystic ovaries.

Recent data suggest that steroidogenic enzyme messenger ribonucleic acids (mRNAs) may be overexpressed in thecal cells, and LH receptors may be prematurely expressed in granulosa cells in women with polycystic ovaries. The purpose of this study was to determine whether there is abnormal gene expression in thecal and granulosa cells from polycystic ovaries. Ovarian tissue specimens were obtained from 12 women with PCOS and 24 regularly cycling control women. The granulosa cells and the theca interna were microdissected from individual follicles. LH receptor, steroidogenesis acute regulatory protein (StAR), cholesterol side-chain cleavage cytochrome P450 (CYP11A), and 17alpha-hydroxylase/C(17-20) lyase cytochrome P450 (CYP17) mRNAs were measured by RT-PCR. There was no difference between 3- to 7-mm control follicles and dominant follicles with respect to LH receptor mRNA expression in either thecal or granulosa cells. CYP11A and CYP17 mRNAs were higher in thecal cells from 3- to 7-mm follicles than in dominant follicles, but StAR expression was not different. In granulosa cells, StAR and CYP11A mRNA expression was higher in dominant follicles than in 3- to 7-mm follicles. The mean levels of LH receptor, StAR, CYP11A, and CYP17 mRNA expression were higher in thecal cells from PCOS follicles than in size-matched control follicles. In granulosa cells, the mean levels of LH receptor and CYP11A, but not StAR, mRNA expression were higher in PCOS than in control follicles. These data demonstrate that regulatory protein and steroidogenic enzyme mRNAs are overexpressed in thecal and granulosa cells from polycystic ovaries and support the conclusions that the thecal cells are hyperstimulated and the granulosa cells may be prematurely luteinizing.

[Luteal and follicular phase inhibin A and B in regularly cycling women]. / Inhibina A oraz inhibina B fazy follikularnej oraz lutealnej cyklu u kobiet w piatej dekadzie zycia.

OBJECTIVE: To investigate the level of inhibin A nad B in luteal and follicular phase in women of reproductive age. PATIENTS: Seventy women 39-52 years of age with regular menstrual cycle. INTERVENTIONS: Blood samples obtained on days 3-8 and on days 22-25 of menstrual cycle were assayed for FSH, estradiol, inhibin A, inhibin B. RESULTS: Luteal and follicular phase inhibin B was correlated inversely with age. Luteal phase inhibin A was correlated inversely with follicular phase FSH. CONCLUSION: Main form of inhibin in follicular phase of the cycle is inhibin B and in luteal phase inhibin A. Inhibin B can be potential marker of ovarian aging.

[Aromatase (P450AROM) mRNA expression in normal, hyperplastic and malignant endometrium and aromatase activity in endometrial cancer tissue culture]. / Ekspresja genu P450AROM w prawidlowym, rozrostowym i nowotworowym endometrium oraz aktywnosc aromatazy w hodowli skrawków raka endometrium.

Aromatase (P450AROM) is the enzyme complex with converts testosterone to estradiol and androstendione to estrone. This enzyme was detected in various normal tissues and uterine pathology such as uterine myoma, endometrial cancer and endometriosis. The aim of the study was to estimate expression of P450AROM messenger ribonucleic acid (mRNA) in normal, hyperplastic and malignant endometrium, and the ability to convert androstenedione to estrone by endometrial cancer tissue. Normal endometrium was obtained from 16 (12 proliferative phase, 4 secretory phase) regularly cycling women after hysterectomy for myomas, hyperplastic endometrium (n = 5) and endometrial cancer (n = 5) from postmenopausal women. The ability to convert androstenedione to estrone was estimated in 16 cases of endometrial cancer in postmenopausal women. P450AROM mRNA was measured by a quantitative assay based on reverse transcribing the mRNA into cDNA with reverse transcriptase (RT) then amplification of the cDNA using the polymerase chain reaction (PCR). The mean (+/- SEM) expression of aromatase gene in proliferative endometrium was 84.4 +/- 14.0 pg mRNA/microgram DNA and in secretory endometrium 200.3 +/- 87.8 pg mRNA/microgram DNA. The mean (+/- SEM) P450AROM mRNA expression in endometrial hyperplasia was 92.9 +/- 17.8 pg mRNA/microgram DNA, in endometrial cancer was 14.3 +/- 7.7 pg mRNA/microgram DNA. Androstenedione to estrone conversion in endometrial cancer tissue culture was 252.5 +/- 91 fmol/g tissue/h. Our data confirm that human normal, hyperplastic and malignant endometrium do express P450AROM mRNA and that aromatase activity is present in endometrial cancer tissue.

Evaluation of matrix metalloproteinases-1 and -3 concentrations in the tunica albuginea, the apical wall of atretic follicles and the corpus luteum of normal human ovaries.

Matrix metalloproteinases-1 (MMP-1) and -3 (MMP-3) are proteolytic enzymes involved in remodeling the ovarian extracellular matrix throughout the menstrual cycle. The aim of the present study was to evaluate the tissue concentrations of MMP-1 and MMP-3 in the apical wall of atretic follicles (androstenedione/estradiol ratio > 4), tunica albuginea dissected from the ovarian surface overlying areas devoid of follicles, corpus luteum, and tunica albuginea covering the corpus luteum. After extraction of MMPs from the tissue samples, their concentrations in the extracts were measured by enzyme-linked immunosorbent assays (ELISA). Significantly less MMP-1 was detected in the apical wall of atretic follicles compared to tunica albuginea taken from sites devoid of follicles. These data indicate that atresia is associated with relatively low concentrations of MMP-1 in the apical wall of the follicle. Moreover, there was a negative correlation between the amount of MMP-3 and the diameter of follicle. These data suggest that both MMPs play an important role in the final step of atresia. The amount of MMP-1 in the corpus luteum was several times lower than in the other tissues. This is likely due to stabilization of the extracellular matrix during the period of the corpus luteum maintenance. The concentration of MMP-3 did not differ significantly among the examined tissues.

Interstitial collagenase (MMP-1) activity in human ovarian tissue.

The collagen content and collagenase activity were estimated in human ovarian interstitial tissue devoid of all visible follicles in menstruating, fertile as well as climacteric women. The mean total collagenase activity in ovarian specimens taken during both follicular (n = 10, 3.97 +/- 0.58 U/g wet weight, ww) and luteal phase (n = 10, 3.39 +/- 1.24 U/g ww) of the normal menstrual cycle along with total collagen concentration (184.8 +/- 41.0 vs. 194.4 +/- 30.5 micrograms/mg ww, respectively) did not differ. Total collagenase activity of climacteric gonads (n = 5, 1.55 +/- 0.71 U/g ww) was lower than in specimens collected during both follicular and luteal phase (p = 0.0002 and p = 0.017, respectively). About 23% of the total collagenase activity in follicular phase ovarian extracts and only about 1% in luteal phase ovarian preparations was found in the latent form. The percentage of latent collagenase in ovarian tissue during the follicular phase was negatively correlated with the day of the menstrual cycle (r = -0.93, p = 0.007). Extracellular matrix remodelling in the human ovary can be correlated with the functional status of the follicular unit.

5alpha-reductase activity in women with polycystic ovary syndrome.

The recent demonstration of high concentrations of 5alpha-androstane-3,17-dione in the follicular fluid of polycystic ovaries suggests a potential role for 5alpha-reduced androgens in the etiology of polycystic ovary syndrome (PCOS). The purpose of the present study was to determine whether there is increased 5alpha-reductase activity or messenger ribonucleic acid (mRNA) expression in polycystic ovaries. 5alpha-Reductase 1 and 5alpha-reductase 2 mRNAs were measured in thecal (TC) and granulosa (GC) cells from individual follicles of 18 women with PCOS and 26 regularly cycling control women. Both 5alpha-reductase 1 and 2 mRNA expression was higher in GC than in TC, and 5alpha-reductase 2 mRNA levels were approximately 3-fold higher than 5alpha-reductase 1 mRNA. 5alpha-Reductase 1 and 2 mRNA expression were similar in GC from PCOS and control women, but 5alpha-reductase mRNA was decreased in TC from PCOS follicles. In control women, 5alpha-reductase 2 mRNA was highest in GC from 3- to 5-mm follicles and decreased to undetectable levels in GC from 7-mm follicles. A similar pattern of expression was present in GC from PCOS follicles, but detectable levels of 5alpha-reductase 2 mRNA were present in GC from 7-mm follicles. 5alpha-Reductase activity was measured in whole follicles by measuring the conversion of radiolabeled testosterone to dihydrotestosterone. Kinetic analysis of total 5alpha-reductase activity at physiological pH revealed a Km of 1.46 micromol/L and a maximal velocity of 0.31 nmol/min x mg protein, indicating predominantly type 1 activity. The total 5alpha-reductase activity was approximately 4-fold higher in PCOS follicles than in control follicles. These data demonstrate elevated 5alpha-reductase activity in polycystic ovaries and support the hypothesis that 5alpha-reduced androgens may play a role in the pathogenesis of PCOS.