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The aim of this study was to provide a comprehensive genetic/phenotypic characterization of subjects suffering infertility owing to sperm macrocephaly (n = 3) or globozoospermia (n = 9) and to investigate whether the patients' genetic status was correlated with the alteration of various sperm parameters. AURKC was sequenced in case of sperm macrocephaly while the DPY19L2 status has been analyzed by multiple approaches including a novel qPCR-based copy number assay in case of globozoospermia. Globozoospermic patients were also analyzed for SPACA1, a novel candidate gene herein tested for the first time in humans. The effect of the patients' genetic status was interrogated by implementing the molecular screening with the characterization of several sperm parameters: (i) routine sperm analysis, integrated with transmission electron microscopy; (ii) sperm fluorescent in situ hybridization (FISH) analysis; (iii) sperm DNA fragmentation (DF) analysis. Moreover, for the first time, we performed microsatellite instability analysis as a marker of genome instability in men with sperm macrocephaly and globozoospermia. Finally, artificial reproductive technology (ART) history has been reported for those patients who underwent the treatment. Macrocephalic patients had an AURKC mutation and >89% tetraploid, highly fragmented spermatozoa. DPY19L2 was mutated in all patients with >80% globozoospermia: the two homozygous deleted men and the compound heterozygous showed the severest phenotype (90-100%). The newly developed qPCR method was fully validated and has the potential of detecting also yet undiscovered deletions. DPY19L2 status is unlikely related to FISH anomalies and DF, although globozoospermic men showed a higher disomy rate and DF compared with internal reference values. No patient was mutated for SPACA1. Our data support the general agreement on the negative correlation between macro/globozoospermia and conventional intracytoplasmic sperm injection outcomes. Microsatellites were stable in all patients analyzed. The comprehensive picture provided on these severe phenotypes causing infertility is of relevance in the management of patients undergoing ART.
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Infertilidad Masculina/complicaciones , Espermatozoides/anomalías , Humanos , Hibridación Fluorescente in Situ , Masculino , Microscopía Electrónica de Transmisión , Espermatozoides/ultraestructuraRESUMEN
'Prostatitis-like symptoms' (PLS) are a cluster of bothersome conditions defined as 'perineal and/or ejaculatory pain or discomfort and National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) pain subdomain score ≥4' (Nickel's criteria). PLS may originate from the prostate or from other portions of the male genital tract. Although PLS could be associated with 'prostatitis', they should not be confused. The NIH-CPSI is considered the gold-standard for assessing PLS severity. Although previous studies investigated the impact of prostatitis, vesiculitis or epididymitis on semen parameters, correlations between their related symptoms and seminal or scrotal/transrectal colour-Doppler ultrasound (CDU) characteristics have not been carefully determined. And no previous study evaluated the CDU features of PLS in infertile men. This study was aimed at investigating possible associations among NIH-CPSI (total and subdomain) scores and PLS, with seminal, clinical and scrotal/transrectal CDU parameters in a cohort of males of infertile couples. PLS of 400 men (35.8 ± 7.2 years) with a suspected male factor were assessed by the NIH-CPSI. All patients underwent, during the same day, semen analysis, seminal plasma interleukin 8 (sIL-8, a marker of male genital tract inflammation), biochemical evaluation, urine/seminal cultures, scrotal/transrectal CDU. PLS was detected in 39 (9.8%) subjects. After adjusting for age, waist and total testosterone (TT), no association among NIH-CPSI (total or subdomain) scores or PLS and sperm parameters was observed. However, we found a positive association with current positive urine and/or seminal cultures, sIL-8 levels and CDU features suggestive of inflammation of the epididymis, seminal vesicles, prostate, but not of the testis. The aforementioned significant associations of PLS were further confirmed by comparing PLS patients with age-, waist- and TT-matched PLS-free patients (1 : 3 ratio). In conclusion, NIH-CPSI scores and PLS evaluated in males of infertile couples, are not related to sperm parameters, but mainly to clinical and CDU signs of infection/inflammation.
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Infertilidad Masculina/diagnóstico por imagen , Dolor Pélvico/complicaciones , Prostatitis/diagnóstico por imagen , Prostatitis/epidemiología , Adolescente , Adulto , Epidídimo/inmunología , Epidídimo/patología , Epididimitis , Humanos , Inflamación/inmunología , Interleucina-8/análisis , Masculino , Persona de Mediana Edad , Próstata/inmunología , Próstata/patología , Prostatitis/diagnóstico , Estudios Retrospectivos , Semen , Análisis de Semen , Vesículas Seminales/inmunología , Vesículas Seminales/patología , Encuestas y Cuestionarios , Testículo/inmunología , Testículo/patología , Testosterona/sangre , Ultrasonografía , Adulto JovenRESUMEN
OBJECTIVES: Previous studies concerning ultrasound evaluation of the seminal vesicles (SV) were performed on a limited series of subjects, and considered few parameters, often only before ejaculation and without assessing the patients' sexual abstinence. The aim of this study was to evaluate the volume and the emptying characteristics of the SV and their possible correlations with scrotal and transrectal ultrasound features. METHODS: The SV of 368 men seeking medical care for couple infertility were evaluated by ultrasound. All patients underwent, during the same ultrasound session, scrotal and transrectal evaluation, before and after ejaculation, and the ejaculate was subjected to semen analysis. A new parameter, SV ejection fraction, calculated as: [(SV volume before ejaculation - SV volume after ejaculation)/SV volume before ejaculation] × 100, was evaluated. RESULTS: After adjusting for sexual abstinence and age, both pre-ejaculatory SV volume and SV ejection fraction were positively associated with ejaculate volume. As assessed by receiver operating characteristic curve, a cut-off for SV ejection fraction of 21.6% discriminates subjects with normal ejaculate volume (≥1.5 ml) and pH (≥7.2 ml) with both sensitivity and specificity equal to 75%. Subjects with SV ejection fraction of <21.6% more often had a higher post-ejaculatory SV volume and ejaculatory duct abnormalities. Furthermore, a higher post-ejaculatory SV volume was associated with a higher prostate volume and SV abnormalities. Higher epididymal and deferential diameters were also detected in subjects with a higher post-ejaculatory SV volume or reduced SV ejection fraction. No association between SV and testis ultrasound features or sperm parameters was observed. Associations with SV ejection fraction were confirmed in nested 1:1 case-control analysis. CONCLUSIONS: The SV contribute significantly to the ejaculate volume. A new parameter, SV ejection fraction, could be useful in assessing SV emptying. A SV ejection fraction of <21.6% was associated with prostate-vesicular and epididymal ultrasound abnormalities.
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Infertilidad Masculina/diagnóstico por imagen , Vesículas Seminales/diagnóstico por imagen , Adulto , Estudios de Casos y Controles , Estudios de Cohortes , Eyaculación , Conductos Eyaculadores/diagnóstico por imagen , Epidídimo/diagnóstico por imagen , Humanos , Masculino , Próstata/diagnóstico por imagen , Escroto/diagnóstico por imagen , Análisis de Semen , UltrasonografíaRESUMEN
BACKGROUND: Obesity is associated with a systemic, low-grade inflammatory state. Although the relationship between obesity and semen parameters or prostate diseases has been previously investigated, the association between body mass index (BMI), prostate inflammatory diseases and color- Doppler ultrasound (CDU) of the male genital tract (MGT) has been poorly studied. AIM: To evaluate the association between BMI and CDU features of the MGT, signs and symptoms of prostate inflammation, semen parameters. MATERIALS/SUBJECTS AND METHODS: We studied 222 men seeking medical care for couple infertility. According to the World Health Organization classification, subjects were divided into 3 groups: normal weight (no.=131, BMI=18.5-24.9 kg/m2), overweight (no.=71, BMI=25.0-29.9 kg/m2), obese (no.=20, BMI≥30.0 kg/m2). All patients underwent simultaneous testosterone evaluation and seminal analysis, including interleukin 8 (sIL-8), along with scrotal and transrectal CDU, before and after ejaculation. Prostatitis symptoms were evaluated by National Institutes of Health- Chronic Prostatitis Symptom Index questionnaire. RESULTS: After adjusting for age and testosterone levels, higher BMI was significantly related to higher prostate volume and several CDU features of the prostate, including macro-calcifications, inhomogeneity, higher arterial peak systolic velocity (the latter adjusted also for blood pressure), but not with abnormalities of testis, epididymis, seminal vesicles. Furthermore, higher BMI and BMI class were significantly related to higher sIL-8, a reliable surrogate marker of prostate inflammatory diseases, even after adjustment for age. Conversely, no associations among BMI, clinical symptoms of prostatitis or semen parameters were observed. CONCLUSIONS: Subjects with higher BMI might develop CDU and biochemical signs suggestive of prostate inflammation, although not clinically overt.
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Índice de Masa Corporal , Infertilidad/diagnóstico por imagen , Interleucina-8/análisis , Próstata/diagnóstico por imagen , Prostatitis/diagnóstico , Semen/química , Adolescente , Adulto , Humanos , Infertilidad/etiología , Masculino , Persona de Mediana Edad , Obesidad/complicaciones , Próstata/patología , Testosterona/sangre , Ultrasonografía Doppler en ColorRESUMEN
This study was aimed at evaluating the association between seminal plasma interleukin-8 (sIL-8) and colour-Doppler ultrasound (CDU) characteristics of the male genital tract in a series of patients fulfilling the criteria of male accessory gland infections (MAGI). Of 250 subjects seeking medical care for couple infertility, 79 (mean age: 36.4 ± 7.5 years) met the criteria of MAGI and scored higher than the rest of the sample on the National Institutes of Health-Chronic Prostatitis Symptom Index score. All patients underwent simultaneous hormone evaluation and seminal analysis (including sIL-8), along with scrotal and transrectal CDU before and after ejaculation. After adjusting for age, sIL-8 in patients with MAGI was significantly related to several abnormal semen and CDU parameters. In particular, leucocytospermia was closely associated with sIL-8. Ejaculate volume, unlike other semen or hormonal parameters, was negatively associated with sIL-8. When scrotal CDU was performed, sIL-8 was positively related to CDU inhomogeneous, hypo-echoic, hyper-echoic epididymis and to epididymal calcifications. In addition, a positive correlation among sIL-8, hyperaemic epididymis and an increased size of epididymal tail was found. When transrectal CDU was performed, an association among sIL-8 and hyper-echoic seminal vesicles, dilated ejaculatory ducts and duct calcifications was also observed. Finally, sIL-8 was positively related to prostate CDU abnormalities such as calcifications, inhomogeneous/hypo-echoic texture, hyperaemia and high arterial blood flow. No association was found with testis parameters. In conclusion, sIL-8 levels in patients with MAGI are associated with several parameters and CDU abnormalities of epididymis, seminal vesicles, ejaculatory ducts and prostate, but not of the testis. Furthermore, sIL-8 positively correlates with CDU signs of ejaculatory duct inflammatory subobstruction.
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Infertilidad Masculina/metabolismo , Interleucina-8/metabolismo , Semen/metabolismo , Adulto , Ensayo de Inmunoadsorción Enzimática , Humanos , Infertilidad Masculina/diagnóstico por imagen , Masculino , Ultrasonografía DopplerRESUMEN
Endometrial cancer (EC) is a hormone-dependent cancer that currently represents the most frequent malignancy of the female reproductive tract. The involvement of steroid hormones in EC etiology and progression has been reported. More recently, gonadotropins, and, in particular LH/hCG, are emerging as novel regulators of tumor progression. In the present review, we discuss the role of the LH/hCG axis (i.e. LH/hCG and its receptors, LH/hCG-R) in both gonadal and nongonadal tissues, in physiological and neoplastic conditions. In cancer cells, LH/hCG mainly controls cell proliferation and apoptosis. In particular, in EC LH/hCG improves cell invasiveness, through a mechanism which involves the LH/hCG-R, which in turn activate protein kinase A and modulate integrin adhesion receptors. Indeed, the LH/hCG-R mRNA is expressed in primary ECs and this expression correlates with LH/hCG-induced cell invasiveness in vitro. These results lead to hypothesize that recurrent and metastatic ECs, which express LH/hCG-R, could benefit from therapies aimed at decreasing LH levels, through Gn-RH analogues. Hence, the LH/hCG axis could represent a prognostic factor and a new therapeutic target in EC.
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AIM: The aim of the work was to compare two different techniques, local anaesthesia versus continuous intravenous administration of remifentanil for oocyte retrieval. The impact on the medical assisted procreation (PMA) procedures in terms of number and quality of oocytes collected was evaluated. METHODS: The experimental design was a retrospective study which compared two different techniques, local anaesthesia versus continuous intravenous administration of remifentanil. 548 women with the same infertility duration and ovarian reserve were classified in two groups of 274 women each, that underwent to oocytes retrieval with intravenous infusion of remifentanil (first group) or with local anesthesia (second group). RESULTS: The analysis showed that the intravenous infusion of remifentanil doesn't interfere in the quality of oocytes retrieved and embryo score. CONCLUSION: The administration of intravenous remifentanil makes easier the pick-up of oocytes because women had no pain during the procedure. In this way, it was possible to recover more oocytes and to verify that the drug doesn't interfere with the exitus of the techniques. For these reasons we decided to continue in using intravenous infusion of remifentanil for the retrieval of oocytes.
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Alfentanilo/farmacología , Anestesia Local/métodos , Fertilización In Vitro , Piperidinas/farmacología , Propofol/farmacología , Técnicas Reproductivas Asistidas , Adulto , Alfentanilo/administración & dosificación , Femenino , Humanos , Infusiones Intravenosas , Oocitos/efectos de los fármacos , Inducción de la Ovulación , Piperidinas/administración & dosificación , Propofol/administración & dosificación , Remifentanilo , Estudios RetrospectivosRESUMEN
Currently, different approaches are used to select oocytes for in vitro fertilization (IVF) procedures, but they do not assure a significant association with the pregnancy outcome. Since several studies have proposed the expression of HLA-G antigens in early embryos to be a possible marker of elevated implantation rate, we have investigated the presence of soluble HLA-G molecules in 50 follicular fluids (FFs). The results have shown soluble HLA-G antigens (sHLA-G) in 19/50 (38%) FFs. Furthermore, we have related the presence of sHLA-G molecules in FFs to detection of the soluble antigens in culture supernatants of the corresponding fertilized oocyte, evidencing a significant relationship (p=1.3 x 10(-6); Fisher exact p-test). Specific ELISA and Western blot approaches identified both HLA-G5 and soluble HLA-G1 molecules in FFs while immunocytochemical analysis indicated polymorphonuclear-like and granulosa cells as responsible for production of sHLA-G1 and HLA-G5 molecules. In contrast, only sHLA-G1 antigens were detected in culture supernatants of fertilized oocytes. Overall, these results suggest a role for sHLA-G molecules in the ovulatory process and propose the FFs analysis for sHLA-G molecule presence as a useful tool for oocyte selection in IVF.
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Fertilización In Vitro/métodos , Líquido Folicular/inmunología , Antígenos HLA/análisis , Antígenos de Histocompatibilidad Clase I/análisis , Western Blotting , Implantación del Embrión , Ensayo de Inmunoadsorción Enzimática , Femenino , Antígenos HLA/fisiología , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/fisiología , Humanos , Inmunohistoquímica , Masculino , Embarazo , Inyecciones de Esperma Intracitoplasmáticas , Cigoto/inmunologíaRESUMEN
AIM: The aim of the study was to compare the clinical results and efficiency of three insemination technique: intraperitoneal insemination (IPI), fallopian sperm perfusion (FSP) and intrauterine insemination (IUI). METHODS: The experimental design was a prospective, randomized trial. A total of 101 homologous insemination cycles were performed in 71 consecutive couples with unexplained or male subfertility. Couples were randomized to receive IPI or FSP or IUI by predefined tables of randomization and each couple was submitted to the same insemination technique. The primary outcome of the study was the achievement of clinical pregnancy. RESULTS: The results of the study underlined firstly that basal couple composition was not statistically different between the three groups. Moreover, no significant difference in clinical pregnancy rate was observed, despite a clearly positive trend for FSP, especially for unexplained infertility. CONCLUSIONS: Our results showed that the three techniques of insemination IUI, FSP and IPI have similar efficacy on the achievement of clinical pregnancy in couples affected by longstanding infertility.
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Infertilidad Masculina , Inseminación Artificial/métodos , Femenino , Humanos , Masculino , Embarazo/estadística & datos numéricos , Estudios ProspectivosRESUMEN
BACKGROUND: In human reproduction, embryo implantation is complex and poorly understood. At present, no single markers are used in routine treatment to assay biochemical functions of the human embryo. Soluble human leukocyte antigen-G (sHLA-G) could be considered a possible marker of embryo developmental potential. It is localized primarily on the extravillous trophoblast, making this antigen a potential mediator of immune interaction at the maternal-fetal interface during gestation. METHODS: Soluble-HLA-G levels were evaluated by an enzyme-linked immunosorbent assay (ELISA) employing monoclonal antibody MEM-G9. It was evaluated in 318 media of single embryo cultures. We correlated the presence of sHLA-G with embryo morphology and the pregnancy obtained in that treatment cycle. RESULTS: No correlation was found between embryo morphology and sHLA-G levels. Pregnancy was observed only when the medium of at least one transferred embryo contained sHLA-G. In 26 out of 66 patients, none of the obtained embryos showed any detectable sHLA-G molecules and no pregnancy occurred. CONCLUSIONS: From our results, we propose sHLA-G as a potential marker of embryo development: the sHLA-G ELISA can be a useful biochemical assay in addition to embryo morphology in embryo selection for transfer in IVF treatment if there are other embryos with the same morphology.
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Embrión de Mamíferos/inmunología , Desarrollo Embrionario/inmunología , Antígenos HLA/metabolismo , Antígenos de Histocompatibilidad Clase I/metabolismo , Biomarcadores/metabolismo , Medios de Cultivo , Transferencia de Embrión , Ensayo de Inmunoadsorción Enzimática , Femenino , Fertilización In Vitro , Antígenos HLA-G , Humanos , Técnicas In Vitro , Masculino , Embarazo , Solubilidad , Inyecciones de Esperma IntracitoplasmáticasRESUMEN
Our purpose was to test the liquid-based thin layer method on the endometrial cytology. One hundred sixty two consecutive patients before the hysterectomy (55 women because of various causes; 107 asymptomatic postmenopausal women because of prolapsed uterus) had the endometrial cytology (all women) and the biopsy (107 postmenopausal women prolapsed uterus affected). Cytohistologic concordance was 98%: all endometrial neoplasms and atypical hyperplasia (10 cases) and 15 of the 18 (83%) simple hyperplasias were diagnosed by thin layer endometrial cytology. In asymptomatic postmenopausal women cytology gave sufficient material for the diagnosis significantively more often than endometrial biopsy (respectively 82% and 24%; p = 0.000).
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Endometrio/patología , Técnicas de Preparación Histocitológica , Enfermedades Uterinas/diagnóstico , Adenocarcinoma/diagnóstico , Adenocarcinoma/patología , Biopsia , Hiperplasia Endometrial/diagnóstico , Hiperplasia Endometrial/patología , Neoplasias Endometriales/diagnóstico , Neoplasias Endometriales/patología , Femenino , Técnicas de Preparación Histocitológica/estadística & datos numéricos , Humanos , Histerectomía , Persona de Mediana Edad , Posmenopausia , Enfermedades Uterinas/patología , Prolapso Uterino/patologíaRESUMEN
Oxytocin (OT) is a neurohypophysial hormone with unclear physiological functions in the male. Several previous studies indicated that OT might have a role in the ejaculatory process, stimulating sperm release from the epididymal storage. In this study we investigated on the presence and function of OT receptor (OTR) in rabbit and human epididymis. By using RT-PCR, Western and binding studies, we found that OTR gene and protein is expressed in the human epididymis and stimulates in vitro contractility. The immunolocalization of OTR suggests that the receptor is not only present in the smooth muscle cells of the human epididymis but also in the epithelial compartment. Experiments performed in rabbit epididymal epithelial (rEE) cells in culture indicate that OT induces the release of an other potent stimulator of epididymal contractility, endothelin-1 (ET-1), Blocking the ET(A) subtype of the ET-1 receptors, by using a specific antagonist (BQ-123), partially counteracts the contractile effect of OT, suggesting positive interactions between the two peptides in regulating epididymal contractility. Finally, to investigate whether an acute OT administration increases sperm release also in humans, we treated oligozoospermic patients with an intravenous bolus of OT (2.5 IU), just before sperm collection. In a small, single blind study, we found that OT almost doubled sperm retrieval when compared with vehicle administration. Our results indicate that OT might have physiological functions also in the male, controlling epididymal motility and sperm progression through the male genital tract.
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Epidídimo/fisiología , Receptores de Oxitocina/fisiología , Adulto , Anciano , Animales , Unión Competitiva , Endotelina-1/metabolismo , Endotelina-1/farmacología , Epidídimo/química , Epidídimo/citología , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Genitales Masculinos/citología , Genitales Masculinos/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Contracción Muscular/efectos de los fármacos , Miocitos del Músculo Liso/citología , Miocitos del Músculo Liso/efectos de los fármacos , Oligospermia/tratamiento farmacológico , Oxitocina/administración & dosificación , Oxitocina/farmacología , ARN Mensajero/metabolismo , Conejos , Receptores de Oxitocina/análisis , Receptores de Oxitocina/genética , Método Simple Ciego , Recuento de Espermatozoides , Distribución TisularRESUMEN
We report the case of a young woman taking a low-dose oral contraceptive (gestodene 0.075mg and ethinylestradiol 0.02mg) in whom we documented by both hormonal assays and sonographic evaluations the occurrence of ovulation, oocyte fertilization and embryo implantation. However, the implantation process did not complete and only a biochemical pregnancy was registered. On the basis of known actions of estroprogestin on endometrium that are not conducive to implantation, it is possible that the pregnancy loss was originated by oral contraceptive's hormonal influence at endometrial level.
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Anticonceptivos Orales/efectos adversos , Implantación del Embrión/efectos de los fármacos , Aborto Espontáneo , Adulto , Gonadotropina Coriónica Humana de Subunidad beta/sangre , Endometrio/diagnóstico por imagen , Etinilestradiol/efectos adversos , Femenino , Humanos , Norpregnenos/efectos adversos , Ovulación , Embarazo , UltrasonografíaRESUMEN
We report here a case of a patient affected by endometrial cancer and treated primarily with leuprolide, the surgical approach being unfeasible due to her compromised conditions. The therapy was continued for more than 6 years, and no progression of the disease was observed. During this period, some histological and immunohistochemical evaluations of the tumour (morphology, grading, proliferation and apoptotic index, E-cadherin expression) were performed. Furthermore, the expression of m-RNA for luteinizing-hormone releasing hormone (LHRH) receptors was determined. The results showed a discrepancy between some biological parameters of the tumour and its clinical characteristics. In fact, despite features suggestive of a progression of the cancer (such as the increase of both tumour grading and proliferating capacity (MIB-1), and a fall in the reparative process (appearance of mutated p53, reduced expression of both bcl-2 and c-erb-2) being detected, neither local invasion nor metastatic lesions were clinically observed. This discrepancy might be due to the maintenance of high levels of E-cadhezin. Moreover, since this tumour was shown to express mRNA for LHRH receptors, new evidence is provided about the favourable impact of LHRH analogue treatment in patients affected by endometrial cancer.
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Adenocarcinoma/tratamiento farmacológico , Antineoplásicos Hormonales/uso terapéutico , Neoplasias Endometriales/tratamiento farmacológico , Leuprolida/uso terapéutico , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Anciano , Biomarcadores de Tumor/metabolismo , Progresión de la Enfermedad , Supervivencia sin Enfermedad , Neoplasias Endometriales/metabolismo , Neoplasias Endometriales/patología , Femenino , Humanos , Proteínas de Neoplasias/metabolismo , ARN Mensajero/metabolismo , Receptores LHRH/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The aim of the present research was to investigate the changes of the sugar residues in the oviduct in the course of ageing in postmenopausal women vs normally menstruating women, by means of lectin histochemistry. Twenty asymptomatic postmenopausal women (48-83 years old) were recruited among patients who underwent a vaginal hysterectomy. Eight normally menstruating women were recruited as controls. Fragments of Fallopian tubes (pars ampullaris) were fixed in 10% formalin and routinely processed. The sections were labelled with HRP-lectins (PNA, SBA, DBA, WGA, Con A, LTA, UEAI). Some sections were pre-treated with neuraminidase prior to staining with HRP-lectins. Among the postmenopausal patients, our histochemical data showed that there was no difference in the localization and distribution of sugar residues of glycoconjugates as detected by various HRP-lectins. Moreover, our results demonstrated that the oviductal epithelium is characterized by apical reactivity in both ciliated and non-ciliated cells. In the course of ageing, the ciliated cells changed their morphology from bathyprismatic to large and rounded shape. ConA lectin reacted intensely with such highly degenerating ciliated cells and could be considered a marker of these cells. The degenerating ciliated cells are also characterized by the absence of sialic acid. In comparison with the sugar residues present in the control group, the oviductal epithelium of postmenopausal women is characterized by the loss of reactivity with DBA, WGA and ConA. Moreover, PNA reactive material was present at the free border of the ciliated and non-ciliated cells. The latter findings were statistically confirmed and could be considered strictly related to the ageing process.
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Trompas Uterinas/crecimiento & desarrollo , Anciano , Anciano de 80 o más Años , Envejecimiento , Metabolismo de los Hidratos de Carbono , Endometrio/anatomía & histología , Endometrio/crecimiento & desarrollo , Trompas Uterinas/anatomía & histología , Trompas Uterinas/metabolismo , Femenino , Histocitoquímica , Peroxidasa de Rábano Silvestre , Humanos , Hidrólisis , Lectinas , Persona de Mediana Edad , Neuraminidasa , Adhesión en Parafina , Posmenopausia/fisiologíaRESUMEN
In a previous study we demonstrated that women with day 3 luteinizing hormone (LH) values < 3 IU/l subjected to controlled ovarian hyperstimulation without pituitary desensitization responded with a lower number of follicles > 15 mm compared to women with a higher basal LH level. The aim of this study was to determine whether in patients with day 3 LH levels < 3 IU/l a further reduction of serum LH concentration by gonadotropin-releasing hormone (GnRH) analog impairs follicular response to follicle stimulating hormone (FSH) and treatment outcome in in vitro fertilization (IVF) cycles. For this purpose we retrospectively studied 249 consecutive women subjected to standard IVF treatment employing pituitary desensitization with buserelin and follicular stimulation with urinary highly purified FSH. The patients were divided into two groups according to their day 3 LH value. The first group (group A) showed day 3 LH levels < 3 IU/l and the second (group B) had day 3 LH levels > 3 IU/l. Group A and B patients did not show statistically significant differences in the ovarian response to FSH, nor in IVF treatment outcome, showing that in FSH treated GnRH analog suppressed cycles, the ovarian responsiveness and IVF outcome do not differ according to basal LH values. However, the high dosage of FSH we employed in group A and B patients could account, at least in part, for this result. Indeed, comparative evaluations with unsuppressed cycles (our previous study) strongly suggest that a reduced ovarian responsiveness to gonadotropins in patients with day 3 LH values < 3 IU/l should be considered in clinical practice.
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Buserelina/uso terapéutico , Fertilización In Vitro , Hormona Luteinizante/sangre , Inducción de la Ovulación/métodos , Adulto , Estradiol/sangre , Femenino , Fármacos para la Fertilidad Femenina/uso terapéutico , Hormona Folículo Estimulante/sangre , Hormona Folículo Estimulante/uso terapéutico , Humanos , Infertilidad Femenina/etiología , Infertilidad Femenina/fisiopatología , Masculino , Ciclo Menstrual , Embarazo , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
HERG K(+)channels, besides contributing to regulate cardiac and neuronal excitability, are preferentially expressed in tumour cell lines of different histogenesis, where their role in the development and maintenance of the neoplastic phenotype is under study. We show here that both herg gene and HERG protein are expressed with high frequency in primary human endometrial cancers, as compared to normal and hyperplastic endometrium. RT-PCR and immunohistochemistry, using specific anti-HERG antibodies developed in our laboratory, were applied to tissue specimens obtained from 18 endometrial cancers and 11 non-cancerous endometrial tissues. herg RNA and HERG protein are expressed in 67% and 82%, respectively, of cancerous, while in only 18% of non-cancerous tissues. In particular, no expression was found in endometrial hyperplasia. Moreover, electrophysiological experiments confirmed the presence of functioning HERG channels on the plasma membrane of tumour cells. On the whole, these data are the first demonstration of the presence of HERG channels in primary human neoplasias, and could candidate HERG as a potential tool capable of marking cancerous versus hyperplastic endometrial growth.
Asunto(s)
Proteínas de Transporte de Catión , Proteínas de Unión al ADN , Neoplasias Endometriales/metabolismo , Endometrio/metabolismo , Canales de Potasio con Entrada de Voltaje , Canales de Potasio/metabolismo , Transactivadores , Anciano , Anciano de 80 o más Años , Bencimidazoles/farmacología , Western Blotting , Canal de Potasio ERG1 , Electrofisiología , Neoplasias Endometriales/genética , Neoplasias Endometriales/patología , Endometrio/patología , Canales de Potasio Éter-A-Go-Go , Femenino , Regulación de la Expresión Génica , Humanos , Hiperplasia , Inmunohistoquímica , Potenciales de la Membrana/genética , Persona de Mediana Edad , Canales de Potasio/genética , ARN/genética , ARN/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sulfanilamidas/farmacología , Regulador Transcripcional ERG , Células Tumorales CultivadasRESUMEN
We studied the effects of luteinising hormone-releasing hormone (LHRH) agonist leuproreline (1 microM for 96 h) and LHRH antagonist cetrorelix on the cell growth of primary cultures from nine human endometrial cancers using the sulphorhodamine colorimetric test. Histological examinations and reverse transcription and polymerase chain reaction amplification (RT-PCR) for LHRH receptors were also performed. The endometrial cancers examined had a medium to high degree of proliferative activity and a low degree of apoptotic power; furthermore, they expressed the LHRH receptor RNA variably, detectable in 71% of cases. The addition of leuproreline or cetrorelix to cell cultures inhibited growth in a statistically significant way compared to untreated control cells; nevertheless, the percentage of cell growth inhibition obtained was very variable. These data suggest that LHRH analogues can exert differential inhibitory effects on the growth of endometrial cancer, which seems to be independent of the expression of specific LHRH receptors.
Asunto(s)
Neoplasias Endometriales/prevención & control , Hormona Liberadora de Gonadotropina/farmacología , Anciano , División Celular/efectos de los fármacos , Neoplasias Endometriales/genética , Neoplasias Endometriales/patología , Femenino , Hormona Liberadora de Gonadotropina/análogos & derivados , Humanos , Leuprolida/farmacología , Persona de Mediana Edad , Posmenopausia , ARN Mensajero/genética , Receptores LHRH/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Soluble human leukocyte antigen (sHLA) class I molecules have been described in all human fluids. These molecules play a significant role in immune function. sHLA has been shown to produce tolerance and to induce apoptosis in cytotoxic alloreactive T cells. They are also present in the supernatant of many cultured cells. Similarly, non-classic HLA class I antigens in soluble form are present in human fluids. Among these, HLA-G is the most important because of its location in fetal tissue that suggests maternal immunological tolerance of the fetal semiallograft. In our present study we show that using two monoclonal antibodies, w6/32 and TP25.99, in the enzyme-linked immunosorbent assay allows the detection of non-classic sHLA class I molecules in the medium from human embryo cultures. The sample were collected from oocytes cultures. Oocyte donors were 11 women attending the in vitro fertilization program. The results showed a significant association (chi2 = 9.66, p = 0.002) between sHLA antigens and the oocyte cleavage rate measured 48 h after fertilization.
Asunto(s)
Antígenos HLA/metabolismo , Antígenos de Histocompatibilidad Clase I/metabolismo , Oocitos/inmunología , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Fase de Segmentación del Huevo/inmunología , Medios de Cultivo , Femenino , Fertilización In Vitro , Humanos , Técnicas In Vitro , Oocitos/crecimiento & desarrollo , SolubilidadRESUMEN
In the clinical management of in-vitro fertilization (IVF) patients it would be very useful to know, before the embryo transfer, whether or not there is a significant chance of pregnancy in that cycle. If low, it would be better to freeze the embryos and postpone the embryo transfer to a subsequent cycle. For this reason, a retrospective study was carried out to investigate the correlations between the serum CA-125 values before embryo transfer and the clinical outcome of that IVF cycle. Women aged <40 years undergoing a complete infertility evaluation including laparoscopy and receiving gonadotrophin-releasing hormone analogue (GnRHa) suppression followed by purified follicle stimulating hormone (FSH) for IVF-embryo transfer were entered into the study. Ninety-seven cycles qualified for evaluation (26 pregnant and 71 non-pregnant cycles). CA-125 concentrations on the day of oocyte retrieval were significantly lower in the pregnant versus non-pregnant cycles in both non-endometriosis and endometriosis patients. To evaluate the existence of a cut-off value of CA-125 which would allow the prediction of a possible pregnancy with sufficient specificity and sensitivity, a receiver operating characteristic curve analysis was performed. This analysis demonstrated the absence of any predictive value of the subsequent pregnancy for CA-125 concentrations. For this reason, and in contrast with previous findings, CA-125 determinations before the embryo transfer in IVF patients do not appear to be a useful tool for clinicians to use in predicting the outcome of IVF in any given cycle.