Fertility in women of late reproductive age: the role of serum anti-Müllerian hormone (AMH) levels in its assessment.

INTRODUCTION: Fertility is referred to the capability for having offspring and can be evaluated by fertility rate. Women's fertility is strictly dependent on individual's age. The fertility peak occurs in the early 20s, and it starts to decline in the third and fourth decades of life (falling sharply after age 35). AIM: The aim of this work is to review the available data concerning fertility in women of late reproductive age, especially the role of serum anti-Müllerian hormone (AMH) levels. RESULTS: There are a lot of factors responsible for decrease of fertility in women of late reproductive age. These factors can be classified as oocyte-dependent (decrease in oocyte quantity and quality) and oocyte-independent (reproductive organs [uterus, oviducts] status and general health). Anti-Müllerian hormone (AMH) is a dimeric glycoprotein of the transforming growth factor-ß (TGF-ß) superfamily produced directly by the ovarian granulosa cells of secondary, preantral, and early antral follicles. It has been used as an ovarian reserve marker since 2002. Anti-Müllerian hormone seems to be the best endocrine marker for assessing the age-related decline of the ovarian pool in healthy women. Evaluation of AMH's predictive value in the naturally aging population is important for counseling women about reproductive planning as well as for treatment planning for women experiencing hormone-sensitive gynecological conditions such as endometriosis and fibroids. CONCLUSIONS: AMH can be considered as an indicator of fertility in late reproductive age women and pregnancy outcome in assisted reproductive technology cycles. AMH can strongly predict poor response in the controlled ovarian stimulation.

The impact of the presence of antithyroid antibodies on pregnancy outcome following intracytoplasmatic sperm injection-ICSI and embryo transfer in women with normal thyreotropine levels.

PURPOSE: The aim of our study was to investigate the impact of anti-thyroid peroxidase antibodies (Anti-TPO) on pregnancy outcome following the in vitro fertilization and embryo transfer (IVF-ET) in general groups and in subgroups divided according to AMH level and age. METHODS: A total of 114 patients positive for anti-thyroid peroxidase antibodies (Anti-TPO+ group) and 495 infertile women negative for anti-thyroid peroxidase antibodies (Anti-TPO- group) undergoing IVF with ICSI from April 2010 to April 2012 were analyzed retrospectively. RESULTS: There were no significant differences in age, BMI, basal FSH, LH, AMH levels and duration of infertility between the two main groups. No significant differences in terms of the days of ovarian stimulation, estradiol level in day 8, total gonadotropin dose, number of oocytes retrieved, available embryos and blastocysts, number of embryos transferred nor in rates of fertilization, implantation, clinical pregnancy, live birth and abortion rate between two main groups were found. The only statistically significant difference among the groups with different anti-TPO antibodies levels was found in basal FSH concentration and BMI. Among the clinical outcomes of IVF with respect to the different anti-TPO levels, the only significant difference was found for the number of oocytes retrieved. Analysis of the baseline parameters in relationship to age categories and AMH levels found significant differences between women positive and negative for thyroid antibodies with respect to basal FSH and LH levels for women >37 years and for basal FSH in AMH <0.6 subgroup. CONCLUSIONS: The present study reveals that patients with anti-TPO antibodies showed no significant differences in fertilization, implantation, pregnancy rates, live birth rates and no higher risk for miscarriage following IVF-ET when compared with those negative for anti-thyroid antibodies.

Microsatellite instability and HPV genotype in Polish women with cervical cancer.

INTRODUCTION: Human papillomaviruses (HPVs) are associated with anogenital cancer. Little is known about the prevalence of microsatellite instability (MSI) in cervical cancer. The aim of this study was to investigate the incidence of microsatellite instability in cervical cancer and to see whether there is a relation between MSI, HPV and clinicopathological characteristics in the study population. RESULTS: Using three assays (pU1M/2R, GP5+/6+ and E6-nested multiplex PCR) HPV was detected in 110 out of 113 patients with histologically confirmed cervical cancer. The presence of MSI was investigated in 95 of the 113 cases using seven microsatellite loci. In total, 12 out of the 95 patients (12.6%) showed MSI. None of clinicopathological parameters showed a significant difference between microsatellite stable and MSI cases. CONCLUSION: In this population of Polish cervical cancer patients, 12.6% showed microsatellite instability. There was no correlation between MSI positivity and clinicopathological parameters and/or survival.

Survival of 231 cervical cancer patients, treated by radical hysterectomy, according to clinical and histopathological features.

PURPOSE OF INVESTIGATION: The purpose of the study was to estimate the five-year survival of cervical cancer patients after radical hysterectomy, taking into account clinical data and histopathological parameters. METHODS: 231 patients with invasive cervical carcinoma were diagnosed, surgically treated--Piver III--and followed-up. Histological examination of specimens was performed according to the British NHS-CSP guidelines. RESULTS: We discovered no statistical significance as regards age at diagnosis, age at menarche and menopause, and number of pregnancies, deliveries and abortions, in relation to survival. We concluded that the clinical stage according to FIGO classification influenced survival. Statistical significances were: Ia2 vs Ib, Ib vs IIa and IIa vs more advanced than IIa. The following histopathological parameters correlated with survival: depth of cervical invasion, primary lesion volume, and parametrial, uterine, vaginal and lymph node involvement. Using Cox's proportional hazards model we found that only lymph node status and FIGO staging were independent parameters correlating with survival and mortality risk in our study. CONCLUSION: Prognostic indexes classifying patients at specific disease stages into different categories of risk should be based on histopathological features listed above. Such indexes are yet to be validated in larger, prospective studies conducted in different patient populations.

An investigation of the potential effect of sperm nuclear vacuoles in human spermatozoa on DNA fragmentation using a neutral and alkaline Comet assay.

Presence of vacuoles and degree of sperm DNA damage are considered to be the basic factors used for the assessment of sperm fertilization capacity. We aimed to investigate the link between these two parameters. According to our knowledge, this is the first study where the Comet assay was used to assess the degree of DNA fragmentation of sperm categorized by Motile Sperm Organelle Morphology Examination (MSOME) Grades. Semen samples from 10 patients were assessed. Spermatozoa were graded into four MSOME groups according to the Vanderzwalmen's criteria. A total of 3930 motile spermatozoa were selected one-by-one using an inverted microscope and transferred onto two different slides. The degree of DNA fragmentation was analyzed by alkaline and neutral Comet assay. Results of the neutral Comet assay showed that Grade I spermatozoa (absence of vacuoles) presented significantly lower dsDNA fragmentation level (mean: 3.13 ± 1.17%) than Grade II (maximum of two small vacuoles; mean: 10.34 ± 2.65%), Grade III (more than two small vacuoles or at least one large vacuole; mean: 23.88 ± 8.37%), and Grade IV (large vacuoles associated with abnormal head shapes or other abnormalities; mean: 36.94 ± 7.78%; p < 0.05). Results of the alkaline Comet assay showed that Grade I spermatozoa had significantly lower DNA (ssDNA + dsDNA) fragmentation level (mean: 8.33 ± 3.62%) than Grade III (mean: 25.64 ± 9.15%) and Grade IV (mean: 40.10 ± 9.10%, p < 0.05), but not significantly lower than Grade II (mean: 12.73 ± 5.06%; p > 0.05). Probably, the vacuoles may be responsible for double strand DNA breaks rather than single strand DNA breaks (only 2.39% spermatozoa in MSOME Grade II, 1.76% in III, and 3.16% in IV has single strand breaks). The results demonstrate that lower MSOME grading correlates with lower sperm DNA fragmentation. Therefore, the observation of sperm nuclear vacuoles using real-time optical microscopy without precise DNA fragmentation examination is not sufficient for optimal sperm selection for intracytoplasmic sperm injection.

Conversion of methanol to hydrocarbons over zeolite ZSM-23 (MTT): exceptional effects of particle size on catalyst lifetime.

A variety of synthetic procedures have been used to obtain zeolite ZSM-23 (MTT) catalysts with crystallite sizes ranging from the micrometer to nanometer scale. When the acidic zeolite is used as a catalyst for the methanol to hydrocarbon (MTH) reaction, the catalytic lifetime is dramatically influenced by the crystallite shape and size.

HPV and histological status of pelvic lymph node metastases in cervical cancer: a prospective study.

AIMS: The purpose of this prospective study was to describe the incidence and distribution pattern of human papillomavirus (HPV) DNA in intraoperative dissected lymph nodes and to relate this to the pathological confirmation of metastasis. METHODS: Samples of primary cervical cancer lesions and dissected lymph nodes were obtained from women undergoing surgical treatment. The presence of HPV DNA was detected by the polymerase chain reaction. RESULTS: Tissue from 79 tumours and 365 lymph nodes was analysed. Metastasis to the lymph nodes was found in 19 cases. Metastasis correlated with the volume of the primary lesion, the depth of cervical and vaginal invasion, and with invasion of the corpus. HPV DNA was found in 60 of the primary lesions and 31 of the lymph nodes. The presence of HPV DNA in the lymph nodes correlated with the volume of the primary lesion and vaginal invasion. CONCLUSIONS: The incidence of HPV DNA in lymph nodes is twice as high as that of histopathologically confirmed metastases. The risk of the presence of HPV DNA and histopathologically confirmed metastases in lymph nodes is related to certain features of the primary tumour.

Lower interleukin-2 and higher serum tumor necrosis factor-a levels are associated with perimenstrual, recurrent, facial Herpes simplex infection in young women.

The aim of this study was to look at a possible relationship between the recurrent perimenstrual dermatosis - facial Herpes simplex infection and the serum concentrations of interleukin-2 (IL-2) and tumor necrosis factor alpha (TNF-alpha). Twenty-one volunteers (19-26 year olds) were examined at five points of the menstrual cycle. Ten volunteers were characterised by recurrent Herpes simplex infection lasting either from the 18th or the 25th day of the menstrual cycle until a few days after menstruation. Eleven young women without symptoms formed the control group. Both groups were similar as regards blood levels of 17beta-estradiol and progesterone. The group with the frequent infectious symptoms was characterised, however, by lower concentrations of IL-2 throughout the whole menstrual cycle, as compared to those without the symptoms. Levels of IL-2 in this group additionally dropped significantly on the 18th and on 25th day of the cycle. Moreover, the group with symptoms was characterised by higher level of TNF-alpha on the 18th day. These changes were found during the menstrual cycle of the women with recurrent herpes infection who however, at the time of the examination were free of the clinical symptoms. There was a similar tendency in both groups towards an increase in the levels of TNF-a around menstruation. Measurement of the other serum pro-inflammatory marker - IL-6 showed higher levels of this cytokine during the menstrual cycle in the group with the clinical symptoms. The results indicate that a decrease of IL-2 together with an increase of TNF-alpha and IL-6 in the serum seem to be related to recurrent perimenstrual Herpes simplex infection.

[The use of biodegradable valtrac-bar rings for intestinal anastomosis in advanced ovarian cancer patients]. / Ocena przydatnosci biofragmentowanych pierscieni dociskowych typu Valtrac-Bar do zespolen jelitowych u chorych na zaawansowanego raka jajnika.

Intestinal anastomoses using Valtrac-Bar biofragmentable rings were performed in 9 advanced ovarian cancer patients presenting with symptoms of acute or subacute intestinal obstruction. Two ileo-transversal anastomoses, six sigmo-rectal and one ileo-ileal ones were performed. A part of sigmoid colon with tumour causing obstruction was resected in seven patients and in two patients a resection of an ileal loop was performed. In two patients a colostomy was necessary. The result of anastomosis with Valtrac-Bar was satisfactory in all patients: no cases of peritonitis, anastomosis leak or fistulae were observed. The treatment of intestinal obstruction allowed for a prolongation and improvement of quality of life of these patients and enabled further treatment with chemotherapy. We conclude that biofragmentable Valtrac-Bar ring is a safe and effective device enabling easy and fast intestinal anastomosis even in patients with inappropriate healing.

[Hormonal profile in infertile men]. / Profil hormonalny pacjentów z meska nieplodnosciá.

Some given sex hormones in patients treated due to marital infertility in this paper is analysed. Testosterone, prolactin, FSH and LH were examined. Males with sperm pattern showing evident pathology into our study were chosen: azoospermia, severe oligospermia, kryptozoospermia.

[Large-scale screening for early diagnosis of gestational diabetes mellitus (GDM)]. / Wczesne rozpoznawanie cukrzycy ciazowej w populacyjnych badaniach przesiewowych.

The aim of study was to evaluate incidence of GDM in Poland. All 1500 pregnant women between 24-28 week's gestation consulted in 4 centers were offered a 50 g oral glucose test (screening test). Capillary blood glucose was measured at 60 min after glucose ingestion. When blood glucose > 140 mg/dl, 75 g OGTT according to WHO criteria was performed. 241 women have abnormal screening test and in 181 cases blood glucose were at range 140-160 mg/dl, in 39 at range 160-180 and 21 were > 180 mg/dl. Only 14 women in the first group (140-160 mg/dl) have diagnosed GDM (7.7%). In second group 24 pregnant women have GDM (61.5%). Overall GDM incidence is shown to be 3.7% (57 women). The mean age for the GDM was 28.8 +/- 0.9 years compared with 26.4 +/- 0.4 years (p < 0.05) uncomplicated pregnancy.

[The use of PCR technique in the evaluation of the frequency of HPV infection in vulvar and cervical carcinoma]. / Zastosowanie techniki PCR do oceny czestosci wystepowania infekcji HPV w rakach sromu i szyjki macicy.

OBJECTIVES: In this study we estimated the frequency of human papillomavirus (HPV) infection in patients with vulvar and cervical carcinoma by polymerase chain reaction (PCR). We took into consideration the age of patients, the HPV infection status and frequency of HPV types. MATERIALS AND METHODS: Vulvar and cervical smears obtained from 88 patients with vulvar and cervical carcinoma, treated in 1997 in II Department of Obstetrics and Gynecology, Medical University in Gdansk were investigated for HPV DNA. We used the multiplex polymerase chain reaction (M-PCR) with four primer pairs: two primer pairs located in the E6, E7 open reading frame (ORF) for HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 52b i 58 and primer par located in L1 ORF and type-specific primer for HPV16 (E6 ORF). We used restriction fragments length polimorphism PCR (RFLP-PCR) method for HPV typing. RESULTS: HPV DNA was detected in 3 (19%) of the 18 patients with vulvar carcinoma, of whom 3 (100%) had HPV16. In cervical carcinoma, DNA HPV identified in 50 (71%) of the 70, of whom 43 (86%) had HPV16, 3 had HPV31, 3 HPV33 and one was positive for HPV52-b. CONCLUSION: We did not find significant differences between the frequency of the HPV infection in different age groups of patients with cervical carcinoma. In vulvar cancer, we found the HPV infections significantly more often in group of patients in the age below 55.

[Estimation of HPV infection presence in cytological smears in a group of asymptomatic women]. / Ocena wystepowania infekcji HPV w badaniach cytologicznych w grupie pacjentek bezobjawowych.

Papanicolau (Pap) smear is the most important screening method in gynaecological oncology. According to EUROGIN and WHO statements it is suggested to perform additionally molecular detection of human papillomavirus (HPV), which is the most important risk factor of cervical cancer. The aim of this study was to estimate the HPV infection prevalence in healthy, asymptomatic women. Pap smears from 255 women were investigated according to The Bethesda Terminology. HPV DNA was detected by PCR method. We were able to detect high "oncogenic risk" HPV types: 16, 18, 31, 33, 35, 39, 45, 52b, 58 and "low oncogenic risk"--HPV6 and 11. We found only 15 cases (5.9%) of HPV DNA presence. Most of them were HPV16--in 5 patients (33.3%) and HPV58--in 3 (20%). HPV detection could be very useful supplement to Pap smears. It allows selecting the patients with high risk of cervical cancer many years before clinical manifestation of the disease.

Comparison of effect of a panel of membrane channel blockers on the proliferative, cytotoxic and cytoadherence abilities of human peripheral blood lymphocytes.

The ability of human peripheral blood lymphocytes (PBMNC) to incorporate tritiated thymidine upon stimulation with concanavalin A, to perform natural cytotoxic (NK) activity and to form two types of spontaneous "rosettes" with sheep red blood cells (SRBC) upon in vitro treatment with membrane channel blockers of different specificity was estimated. All five channel blockers used, i.e., verapamil, nifedipine, diltiazem, saxitoxin and tetraethylammonium exerted a biphasic, dose-dependent effect on the Con A-induced proliferation of PBMNC. Verapamil, saxitoxin and nifedipine significantly increased the percentage of PBMNC forming spontaneously "active" rosettes with the SRBC. All channel modifiers tested for this ability decreased the percentage of PBMNC forming "late" rosettes. Natural killer activity was significantly decreased in the presence of micromolar concentrations of calcium antagonists as well as when the cells were treated with tetraethylammonium. The results of all four parameters tested were also compared with these obtained in a group of patients treated with 300 mg per day verapamil for two weeks preceding the tests. In this case we have found that treatment in vivo significantly decreased the response of PBMNC to mitogen stimulation, but it was without effect on either NK cytotoxicity or rosette formation. Cells of verapamil-treated patients did not show increased response to Con A in the presence of low concentration of the blocker in vitro. Also, they were not able to form more "active" rosettes when treated with 1-100 nM verapamil. We believe that apparent bi-directional pattern of action of calcium, sodium and potassium channel blockers on the proliferation and rosette formation of human PBMNC can be explained by two sets of events producing opposite results. First, we suggest that at respectively low concentrations of the blockers all of them generate uniform modulation of transmembrane electrical potential (depolarization) of lymphocytes. Depolarization promotes cell adhesion and is probably beneficial for early stages of the response to mitogen. At higher concentrations, this effect is overshadowed by other pharmacological actions of the blockers, of mostly inhibitory character.