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OBJECTIVES: To compare results after stimulation with corifollitropin alfa (Elonva) in unselected group of women entering for the first time in in vitro fertilization programme (IVF) with results from Phase III randomized trials with selected groups of women. DESIGN: Prospective study. SETTING: Sanatorium Pronatal, Praha. METHODS: 40 unselected women with adequat ovarian reserve entering for the first time in IVF programme were stimulated with corifollitropin alfa and GnRH antagonists. Avarage age in the study group was 32,8 years (29-42 years), women younger then 36 and less then 60 kg received Elonva 100 µg , all others (age > 36 let, weight > 60 kg) Elonva 150 µg. Five days after egg retrieval one blastocyst was transferred (single embryo transfer - eSET). Our results were compared with the resuls in higly selected groups of women from Phase III randomized trials. RESULTS: After stimulation with corifollitropin alfa and GnRH antagonists on average 10,6 (9,2 ± 4,2) eggs could be retrieved, among them 7,3 (6,6 ± 3,9) were M II oocytes (68,9%) and fertilisation rate was 84,6%. After first embryo transfer ("fresh" embryos and embryos from "freeze all" cycles) 14 pregnancies were achieved (37,8%), three pregnancies were achieved later from transfer of frozen-thawed embryos (cumulative pregnancy rate 45,9%). There were three abortions. No severe hyperstimulation syndrom occured. Our results in unselected group of women stimulated for the first in an IVF programme with corifollitropin alfa are fully comparable with results published in randomized trials with selected group of patiens. CONCLUSION: Corifollitropin alfa in combination with daily GnRH antagonist can be successfully used in normal-responder patients stimulated for the first time in an IVF programmeKeywords: corifollitropin alfa, GnRH antagonists, ovarian stimulation, pregnancy.
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Fármacos para la Fertilidad Femenina/administración & dosificación , Fertilización In Vitro/métodos , Hormona Folículo Estimulante Humana/administración & dosificación , Infertilidad/terapia , Inducción de la Ovulación/métodos , Adulto , Femenino , Hormona Folículo Estimulante Humana/sangre , Humanos , Embarazo , Índice de Embarazo , Estudios Prospectivos , Resultado del TratamientoRESUMEN
OBJECTIVE: To demonstrate successful restoration of ovarian function after ovarian transplantation in young woman with premature ovarian failure after gonadotoxic treatment of breast carcinoma and to discuss its possibilities. DESIGN: Case report. SETTING: Sanatorium Pronatal, Prague. CASE REPORT: We demonstrate successful ovarian transplantation and restoration of ovarian function in 33 years old woman after cryopreservation of ovarian tissue performed before chemotherapy and radiotherapy of breast carcinoma and discuss possibilities of this approach in women diagnosed with cancer and subsequent risk of ovarian silure. CONCLUSION: Cryopreservation of ovarian tissue is one of efficient options to preserve fertility in young patients facing gonadotoxic treatment.
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Antineoplásicos/efectos adversos , Neoplasias de la Mama/tratamiento farmacológico , Criopreservación , Preservación de la Fertilidad/métodos , Ovario/trasplante , Insuficiencia Ovárica Primaria/cirugía , Adulto , Femenino , Humanos , Insuficiencia Ovárica Primaria/inducido químicamente , Trasplante AutólogoRESUMEN
INTRODUCTION: During the 25th symposium of assisted reproduction in Brno was lunch time organised as the lunch table discussion on the selected topics of assisted reproduction. More than 150 specialists reviewed themes related to gynecology and embryology.Discussed topics: Lunch table discussion covered the following topics: (1) Cross-border health care in assisted reproduction; (2) Indication for PGS (preimplantation genetic screening) in the context of actual information; (3) Does ovarian stimulation belong to the ambulance of registering gynecologists? (4) Therapy with clomifen - only for IVF specialists? (5) How and with whom should psychological support be directed during IVF? (6) Stimulation in women with low ovarian reserve; (7) Is basic semen analyses sufficient? (8) Time-lapse systems as relevant markers of embryonic development; (9) How to be oriented with choices of media and consumables in the IVF lab, and (10) "Freeze All" - is this new trend in cryopreservation suitable for all? CONCLUSIONS: Panel conclusions were presented during the afternoon session, which had great attendance, featured lively commentary, and produced some definitive consensus. Certain issues remained inconclusive, and these matters will be the subject of further discussion in the future. Specific summation of all deductions is presented in this paper.
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The guidlines are the recommendation for good laboratory practice in embryological laboratories. In this first part the requirements of the clean environment from the point of view of the oocytes, sperm, and embryos protection against infection and from point of view of the embryological laboratory staff health protection were described.
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Guías de Práctica Clínica como Asunto , Técnicas Reproductivas Asistidas/normas , Femenino , HumanosRESUMEN
AIM OF THE STUDY: To assess the PGD results in couples with robertsonian and reciprocal translocations. DESIGN: Retrospective study. SETTING: Sanatorium Pronatal, Prague, accredited IVF unit. METHODS: 94 infertile couples with translocation (44 couples with robertsonian and 50 couples with reciprocal translocations) were included in the study. The mean woman's age was not different: 33 +/- 4,4 in robertsonian vers. 33 +/- 3.9 in reciprocal translocations. The performance of FISH probes in specific cases was tested on patient's lymfocytes before the treatment was started. After ovarian stimulation (recombinant FSH or hMG + GnRH agonist, "long" protocol) and transvaginal oocyte pick-up, embryo biopsy of a single cell was performed 72 hours after fertilization. After blastomere fixation, translocated chromosomes + chromosomes 13, 18, 21, X and Y were tested using FISH. The maximum of two embryos euploid for detected chromosomes were transferred, supernumerary euploid embryos were frozen. RESULTS: From the total number of 629 embryos, 126 embryos (21.9%) were detected as normal or with balanced translocation--25.2% (68/270) in couples with robertsonian and 16.4% (59/359) with reciprocal translocation. Embryotransfer was performed in 30 cycles (68.2%) in robertsonian and 27 (54%) in reciprocal translocations. 24 pregnancies were achieved--15 (39% per cycle and 50% per ET) for robertsonian and 9 (19% per cycle and 33% per ET) for reciprocal translocation--this difference was statistically significant (p = 0.033). Only one pregnancy in each group ended as abortion. SUMMARY: IVF is a valuable option for couples with infertility problems and translocation. This technique allows in short-term a conception and delivery of a healthy baby with general better prognosis for couples with robertsonian translocation.
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Heterocigoto , Infertilidad/genética , Diagnóstico Preimplantación , Translocación Genética , Femenino , Fertilización In Vitro , Humanos , Recién Nacido , Infertilidad/terapia , Masculino , EmbarazoRESUMEN
OBJECTIVE: To compare clinical and embryological characteristics in donor cycles triggered for final oocytes maturation with Pregnyl 10 000 IU i.m. versus triptorelin 0.2 mg s.c. in the same patients in two sequential stimulation cycles. The aim of the study is to decrease the risk of the development of ovarian hyperstimulation syndrome (OHSS) at high response donors by the replacement of Pregnyl 10 000 IU i.m. vs. triptorelin 0.2 mg s.c. The administration of a single dose of gonadotropin-releasing hormone agonist (triptorelin 0.2 mg s.c.) induces release of LH from the pituitary gland similarly to a spontaneous LH surge. SUBJECT: Prospective cross-over trial. SETTING: Sanatorium Pronatal, Praha. SUBJECT AND METHOD: From August 2009 to July 2010 we analysed 24 stimulation cycles in 12 egg donors treated with GnRH antagonist protocol with recombinant FSH (follitropin beta). We identified patients with more than 15 follicles during examination by transvaginal ultrasound. When at least 3 leading follicles reached 17 mm in diameter we administrated Pregnyl 10 000 IU i.m. for final oocytes maturation and triptorelin 0.2 mg s.c in the subsequent treatment cycle. RESULTS: The primary outcome measure was number of oocytes, proportion mature oocytes and fertilized oocytes. The secondary outcome were duration of FSH stimulation, total dose of gonadotropins and mean daily dose of gonadotropins. Data was analysed by paired t-test. We retrieved 17.2 +/- 8.6 vs. 15.8 +/- 5.3 (ns) oocytes, 12.6 +/- 7.3 vs. 13.0 +/- 5.4 (ns) metaphase II oocytes, proportion of metaphase II oocytes (%) was 73 vs. 83 (ns), number of fertilized oocytes 11.5 +/- 6.7 vs. 11.7 +/- 4.5 (ns), fertilization rate (%) 91 vs. 90 (ns) in Pregnyl's vs. triptorelin's group, resp. Duration of FSH stimulation (days) 12.2 +/- 0.8 vs. 12.4 +/- 0.7 (ns), total dose of gonadotropins (IU) 1744 +/- 277 vs. 1740 +/- 276 (ns), mean daily dose of gonadotropins (IU) 238 +/- 43 vs. 221 +/- 36 (ns), were not statistically different in both groups. CONCLUSIONS: Number of mature oocytes and subsequent embryonic cleavage is comparable to standard hCG treatment. There are no differences in clinical and embryological characteristics in both groups. Only one patient with administration of Pregnyl 10 000 IU i.m. was treated for OHSS grade II by vaginal paracentesis. Administration of triptorelin 0.2 mg s.c. is a safe and effective approach to achieve mature oocytes in egg donation programme, where we do not take care of implantation, which has got some limitations based on several studies.
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Gonadotropina Coriónica/farmacología , Donación de Oocito , Oocitos/efectos de los fármacos , Pamoato de Triptorelina/farmacología , Femenino , Humanos , Síndrome de Hiperestimulación Ovárica/prevención & controlRESUMEN
Calreticulin, upon translocation to the cell surface, plays a critical role in the recognition of tumour cells and in experimentally induced cellular anti-tumour immunity. However, less is known about anti-calreticulin antibodies and their role in malignancies. Using enzyme-linked immunosorbent assay (ELISA), we found immunoglobulin (Ig)A and/or IgG anti-calreticulin antibodies in sera of approximately 63% of patients with hepatocellular carcinoma (HCC), 57% of patients with colorectal adenocarcinoma (CRA) and 47% of patients with pancreatic adenocarcinoma (PACA), while healthy controls, patients with viral hepatitis C and with chronic pancreatitis reached only 2%, 20% and 31% seropositivity, respectively. We found significantly elevated mean levels of IgA anti-calreticulin antibodies (P < 0.001) in patients with HCC (78.7 +/- 52.3 AU, mean +/- standard deviation), PACA (66.5 +/- 30.9 AU) and CRA (61.8 +/- 25.8 AU) when compared to healthy controls (41.4 +/- 19.2 AU). Significantly elevated mean levels of IgG anti-calreticulin antibodies (P < 0.001) were detected in patients with HCC (121.9 +/- 94.2 AU), gall bladder adenocarcinoma (118.4 +/- 80.0 AU) and PACA (88.7 +/- 55.6 AU) when compared to healthy controls (56.7 +/- 22.9 AU). Pepscan analysis revealed a large number of antigenic epitopes of calreticulin recognized by both IgA and IgG antibodies of patients with HCC and PACA, indicating robust systemic immune response. Moreover, significantly elevated levels of antibodies against peptide KGEWKPRQIDNP (P < 0.001) in these patients, tested by ELISA, confirmed the distinct character of antibody reactivity against calreticulin. The high occurrence and specificity of serum anti-calreticulin autoantibodies in the majority of patients with some gastrointestinal malignancies provide the evidence for their possible clinical relevance.
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Adenocarcinoma/inmunología , Anticuerpos Antineoplásicos/inmunología , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Linfocitos B/inmunología , Calreticulina/inmunología , Carcinoma Hepatocelular/inmunología , Neoplasias Colorrectales/inmunología , Neoplasias Hepáticas/inmunología , Proteínas de Neoplasias/inmunología , Neoplasias Pancreáticas/inmunología , Adenocarcinoma/sangre , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antineoplásicos/sangre , Especificidad de Anticuerpos , Autoanticuerpos/sangre , Carcinoma Hepatocelular/sangre , Neoplasias Colorrectales/sangre , Ensayo de Inmunoadsorción Enzimática , Epítopos/inmunología , Femenino , Neoplasias de la Vesícula Biliar/sangre , Neoplasias de la Vesícula Biliar/inmunología , Hepatitis C Crónica/sangre , Hepatitis C Crónica/inmunología , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina A/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Neoplasias Hepáticas/sangre , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/sangre , Pancreatitis/sangre , Pancreatitis/inmunología , Adulto JovenRESUMEN
Pulmonary hypertension is involved in the development of various diseases and therefore it can be caused by several mechanisms from a simple pressure elevation in the pulmonary artery to the serious impairments of pulmonary vessels. The recently increased interest in the problems of pulmonary hypertension results namely from the new therapeutic means for the treatment of pulmonary arterial hypertension and chronic thrombembolic pulmonary hypertension. The algorism of pharmacotherapy results from the test of acute pulmonary vasodilation. Only the patients with positive test are indicated to the treatment with high doses of calcium channel blockers. Patients with negative test receive beside the chronic anticoagulation therapy also a specific pharmacotherapy (prostanoids, antagonists of endotheline receptors, phosphodiesterase 5 inhibitors) with not only vasodilatory but also with antiproliferative and antiaggregatory effects. When all possibilities of pharmacotherapy are exhausted, balloon atrial septostomy or lung transplantation should be considered. It has been shown recently that similar pharmacotheraeutic approaches as they are used in patients with pulmonary arterial hypertension are effective in some cases of other forms of chronic pulmonary hypertension. Method of choice in the treatment of chronic thromboembolic pulmonary hypertension is the pulmonary endarterectomy in patients with surgically curable thrombotic obstruction. In patients who are not suitable for surgical treatment it is necessary to try pharmacotherapy (prostacycline, bosentan, sildenafil) or lung transplantation. Complicated diagnosis and therapy of pulmonary hypertension requires concentrating the treatment into specialized centres with multidisciplinary background and sufficient experience. In the Czech Republic, the care of patients with pulmonary hypertension is concentrated into the Cardio Center of the 2nd Medical Department of the 1st Faculty of Medicine and General Teaching Hospital in Prague and into the Cardio Center of the Institute of Clinical and Experimental Medicine in Prague. Complex care to patients with chronic thromboembolic pulmonary hypertension is given at the Cardio Center of the General Teaching Hospital in Prague, where since September 2004, 99 patients were surgically treated with results comparable to the best similar departments abroad.
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Hipertensión Pulmonar/terapia , Enfermedad Crónica , Cardiopatías/complicaciones , Humanos , Hipertensión Pulmonar/complicaciones , Embolia Pulmonar/complicacionesRESUMEN
Refractory coeliac disease (RCD) is a very rare and dangerous form of CD, in which gluten-free diet loses its therapeutic effect and the damage of intestinal mucosa persists. Because of the adherence to the diet, serological markers of CD [immunoglobulin A (IgA) antibodies against gliadin, tissue transglutaminase (tTG) and endomysium] are often missing in RCD patients. We found substantially elevated levels of IgA anti-calreticulin (CRT) antibodies in the sera of almost all RCD patients tested. These sera were negative for IgA antibodies to gliadin and tTG and only some of them showed IgA antibodies to enterocytes. Analysis of patients' IgA reactivity to CRT fragments (quarters and halves) by Western blotting revealed differences in the specificity of IgA antibodies between RCD and CD patients. We therefore used the Pepscan technique with synthetic overlapping decapeptides of CRT to characterize antigenic epitopes recognized by serum IgA antibodies of RCD patients. Employing this method we demonstrated several dominant antigenic epitopes recognized by IgA antibodies of RCD patients on the CRT molecule. Epitope GVTKAAEKQMKD was recognized predominantly by serum IgA of RCD patients. Our results suggest that testing for serum IgA antibodies against CRT and its selected peptide could be a very useful tool in RCD differential diagnosis.
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Anticuerpos Antiidiotipos/inmunología , Calreticulina/inmunología , Enfermedad Celíaca/inmunología , Inmunoglobulina A/inmunología , Anciano , Anticuerpos Antiidiotipos/sangre , Western Blotting , Calreticulina/sangre , Enfermedad Celíaca/sangre , Enfermedad Celíaca/diagnóstico , Dieta Sin Gluten/efectos adversos , Enterocitos/química , Enterocitos/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Gliadina/sangre , Gliadina/inmunología , Humanos , Inmunoglobulina A/sangre , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Transglutaminasas/sangre , Transglutaminasas/inmunologíaRESUMEN
INTRODUCTION: Pulmonary arterial hypertension (PAH) is a serious primary illness of the pulmonary arterioles, characterised by progressive precapillary pulmonary hypertension. The conventional therapy for this condition is so-called specific pharmacotherapy, which addresses the key mechanisms in the pathophysiology of the illness, making use of drugs from the prostanoid group, endothelin receptor antagonists and phosphodiesterase inhibitors. Treprostinil is a stable analogue of prostacyclin, which can be administered subcutaneously, intravenously or by inhalation. PATIENT SAMPLE AND METHOD: In the centre for pulmonary hypertension in the Second Internal Clinic of Cardiology and Angiology of 1st Faculty of Medicine, Charles University, and the General Teaching Hospital in Prague, 22 patients with PAH (idiopathic PAH, familial PAH, PAH associated with congenital heart disease and PAH associated with systemic connective tissue disease) were treated with trerpostinil, 18 patients with a continuous subcutaneous infusion and 4 patients with a continuous intravenous infusion. The indicators followed were the distance reached in a 6-minute walking test, functional capacity assessed by NYHA classification and mortality. RESULTS: The patients for whom treprostinil treatment was indicated had an average pressure in the right atrium of 11.9 +/- 4.2 mm Hg, average pressure in the pulmonary artery of 56.8 +/- 10.7 mm Hg, a cardiac index of 1.78 +/- 0.25 l/min/m2 and a total pulmonary resistance of 16.26 +/- 4.48 WU. 15 patients were functionally NYHA III and 7 patients were NYHA IV. The average distance achieved in a 6-minute walk test before the start of treatment was 326 +/- 83 m. When treated with gradually increasing doses of treprostinil the distance achieved in the 6-minute walk test improved. After 6 months, the group that received subcutaneous treatment had extended their distance to 359 m, after 12 months it was 393 m, after 24 months 447 m and after 36 months 494 m. After 6 months, the group that received intravenous treatment had extended their distance to 473 m, which increased to 451 m after 12 months and 489 m after 24 months. Functional capacity also improved. In total 5 patients were unable to tolerate the subcutaneous infusion, of whom 3 were placed on intravenous treprostinil and 2 on oral bosentan. 7 of the patients died in the period examined (31.8%). CONCLUSION: Treprostinil improves symptoms and hemodynamics for PAH patients and reduces mortality.
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Antihipertensivos/administración & dosificación , Epoprostenol/análogos & derivados , Hipertensión Pulmonar/tratamiento farmacológico , Adulto , Anciano , Epoprostenol/administración & dosificación , Tolerancia al Ejercicio , Femenino , Humanos , Hipertensión Pulmonar/fisiopatología , Bombas de Infusión , Infusiones Intravenosas , Masculino , Persona de Mediana Edad , Circulación PulmonarRESUMEN
We examined seven patients with lichen planus pigmentosus (LPP) clinically and microscopically. Clinically, all patients had a striking predominance of lesions in an intertriginous location, with most of them in the axillae. Microscopically, two biopsies were of significance. Except for the regressive lichen planus, which is usual in LPP, the active inflammatory phase was also present. In these biopsies the very intensive hydropic degeneration of basal keratinocytes was combined with the absence of compensatory increased proliferation of keratinocytes, i.e. without acanthosis. The short duration of this process probably led to the quick transformation into a long noninflammatory regressive phase with incontinence of the pigment. These specific morphogenetic dynamics are possibly why most of the morphs of LPP present as brown, non-pruritic, small inflammatory macules. Because of the highly characteristic inverse location of the lesions in our patients we propose the designation LPP-inversus for this variant of the disease.
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Liquen Plano/patología , Anciano , Axila , Biopsia con Aguja , Femenino , Ingle , Humanos , Hiperpigmentación/patología , Inmunohistoquímica , Liquen Plano/diagnóstico , Masculino , Persona de Mediana Edad , Pronóstico , MuestreoRESUMEN
Calreticulin (CRT), a high-affintiy calcium binding protein and chaperone, was recently identified as one of the targets of autoantibodies in coeliac disease. We evaluated the level of IgA and IgG antibodies to CRT in sera from patients with coeliac disease and various autoimmune diseases. The level of antibodies to gliadin (shown previously to cross-react with CTR), isolated enterocytes and tissue transglutaminase were determined for comparison. The mean level of IgA antibodies to CRT was significantly higher (P< 0.001) in sera from coeliac patients with active disease (139.9+/-11.2 AU/+/-SE) than in healthy controls (20.9+/-1.7 AU). In sera of patients with systemic lupus erythematosus (SLE), insulin dependent diabetes mellitus (IDDM), multiple sclerosis (MS) and autoimmune thyroiditis (AT) or inflammatory bowel disease (IBD) the mean level (25.8+/-3.7 to 38.1+/-5.6 AU) did not exceed the cut-off value. A low level of these antibodies, however, was detected in some sera of patients with MS and IBD. The level of IgG anti-CRT antibodies was increased in coeliac patients (mean 125.4+/-8.0 AU, P< 0.001) when compared to that in healthy controls (33.9+/-2.3 AU). The IgG anti-CRT antibodies were also detected in about 30% of SLE patients sera (54.1+/-3.6 AU, P< 0.001), but the mean level reached only half that detected in coeliac patients.
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Autoanticuerpos/sangre , Enfermedades Autoinmunes/inmunología , Proteínas de Unión al Calcio/inmunología , Enfermedad Celíaca/inmunología , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Ribonucleoproteínas/inmunología , Adolescente , Adulto , Anciano , Western Blotting , Calreticulina , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Humanos , Persona de Mediana EdadRESUMEN
Adhesive interactions between endothelium and circulating cells are crucial for the development of inflammatory reactions. We found significantly higher serum levels of soluble intracellular adhesion molecule-1 (sICAM-1, 492.5 +/- 22.1 ng/ml) in patients with active celiac disease (including IgA-deficient patients) than in patients on a gluten-free diet (335.7 +/- 20.0 ng/ml) (P < 0.001) and healthy controls (207.4 +/- 11.2 ng/ml) (P < 0.001). The concentration of soluble E-selectin in sera from celiac patients (37.2 +/- 3.4 ng/ml) was also higher (P < 0.001) than in sera from healthy controls (15.5 +/- 0.7 ng/ml) but, in contrast to sICAM-1, it remained high in the patients after treatment (30.2 +/- 2.7 ng/ml). Interestingly, the concentration of circulating soluble interleukin-2 receptors, molecules indicating lymphocyte activation, was only increased in sera from patients with active celiac disease (2943.0 +/- 214.1 pg/ml), and the level in sera from treated patients and healthy controls was comparable (1936 +/- 349 and 1416 +/- 111.7 pg/ml). The elevated serum level of soluble cell adhesion molecules could be used as a supplementary, noninvasive procedure for monitoring intestinal immune reactions.
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Enfermedad Celíaca/sangre , Selectina E/sangre , Molécula 1 de Adhesión Intercelular/sangre , Receptores de Interleucina-2/sangre , Adolescente , Adulto , Enfermedad Celíaca/dietoterapia , Niño , Preescolar , Femenino , Humanos , Lactante , MasculinoRESUMEN
OBJECTIVE: The study of zona free oocytes fertilization using intracytoplasmic sperm injection (ICSI) with the aim to increase the pregnancy rate in the assisted reproduction. DESIGN: Pilot study. SETTING: Assisted Reproduction Centre, Gynecology Obstetrics Department of the 1st Medical Faculty and General Faculty Hospital, Charles University, Prague. METHODS: Zona free oocytes were fertilized using ICSI and cultured in standard conditions in vitro for 5 days until the blastocyst stage. Blastocysts were cryopreserved using 8% glycerol. RESULTS: The fertilization process of zona free oocyte was physiological, oocytes expulsed the polar body and in standard time, i.e. 18 hour post insemination, two distinct, morphologically normal pronuclei were apparent in cytoplasm. During early cleavage, from the first to the third day of in vitro culture, the dynamics of cell division was normal, the three-dimensional arrangement of cells was more flattened than in normal, zona intact embryo. The fourth and fifth day of culture, the morphological appearance of embryo corresponded to normal development. CONCLUSION: The fertilization of zona free oocytes using ICSI can give morphologically normal zygote and after 5 days culture in vitro these zygotes can develop into blastocyst stage. This method can increase the pregnancy rate in patients with repeatedly low number of oocytes or with defects of zona pellucida.
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Infertilidad/terapia , Oocitos , Inyecciones de Esperma Intracitoplasmáticas , Zona Pelúcida , Adulto , Femenino , Humanos , Proyectos Piloto , EmbarazoRESUMEN
We analyzed the immune response to gliadin in suckling rats and rats hand-fed with an artificial milk formula, an animal model of gluten enteropathy. Animals of both groups were intragastrically given either gliadin or albumin (control animals) or gliadin from birth till day 55. When compared to the controls, spleen lymphocytes from both groups of gliadin-treated rats cultivated in vitro exhibited a significant increase of spontaneous 3H-thymidine incorporation. Moreover, the proliferation of spleen and mesenteric lymph node (MLN) lymhocytes from both groups of gliadin-treated suckling and hand-fed rats was specifically increased by the in vitro gliadin challenge. Spleen B cells from gliadin-treated rats spontaneously produced higher amounts of gliadin-specific antibodies than those from the controls, however, in vitro stimulation by gliadin caused no further increase in antibody production. Apoptotic DNA fragmentation in MLN cells was higher in gliadin-treated rats than in albumin-treated ones, independently of the milk diet during the suckling period.
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Animales Lactantes , Formación de Anticuerpos , Gliadina/inmunología , Inmunidad Celular , Animales , Anticuerpos/análisis , Apoptosis , Linfocitos B/inmunología , Enfermedad Celíaca/inmunología , División Celular , Fragmentación del ADN , Dieta , Alimentos Formulados , Gliadina/administración & dosificación , Ganglios Linfáticos/citología , Activación de Linfocitos , Mesenterio , Leche , Ratas , Ratas Wistar , Bazo/citologíaRESUMEN
In the Centre for Assisted Reproduction of the Second Gynaecological and Obstetric Clinic, First Medical Faculty, Charles University and General Hospital Prague, during the period from 1993-1997, 900 couples were examined on account of sterility. In 8%, i.e. 72 couples the cause was not elucidated. The female patients were divided into groups by age, parity and period of sterility. In these groups the cumulative conception (CCR) was compared as well as the fecundity per cycle after three months without treatment, after three cycles of intrauterine insemination (IUI) and after three cycles of in vitro fertilization (IVF) or intracytoplasmic sperm injection into the oocyte (ICSI). In the group of patients with sterility shorter than three years, the cumulative conception rate after three months without treatment was 34.8%, after the subsequent three cycles of IUI 27.3% and after three cycles of IVF/ICSI 66.7%. In women with sterility extending over more than three years these results were significantly lower: without treatment CCR was 12.5%, when IUI was used it was 16.1% and after IVF/ICSI 40.5%. There were also marked differences in the age groups. Based on these results the authors elaborated an optimal individual therapeutic pattern. Treatment can be postponed by three months in women under 30 years and those where sterility persists for less than three years. If they do not become pregnant, 3-4 IUI cycles should be attempted. Even women where IUI fails have a fair chance to become pregnant by IVF ET. In women above 30 years with sterility lasting more than three years there the chance that they will become pregnant without treatment is only when secondary sterility is involved. IUI increases the probability of pregnancy in this group only slightly. If during stimulation before IUI more than four follicles grow, it is advisable to convert these cycles to IVF. In women above 39 years the authors recommend IVF or ICSI resp. as the method of first choice.
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Infertilidad Femenina/terapia , Adulto , Femenino , Fertilización In Vitro , Humanos , Inseminación Artificial , Embarazo , Inyecciones de Esperma IntracitoplasmáticasRESUMEN
OBJECTIVE: To review the current status and new trends in assisted reproduction. DESIGN: The literature on intrauterine insemination, IVF, ICSI, MESA/TESE, preimplantation genetic diagnosis, cryopreservation, in vitro maturation, cloning and stem cells. SETTING: Assisted Reproduction Centre, Ist Medical Faculty of Charles University Prague and General Hospital Prague, Apolinárská 18, 128,00, Prague 2, Czech Republic. RESULTS: IUI, IVF, ICSI also in combination with MESA/TESE are widely used relatively high effective procedures. According to Czech Republic Assisted Reproduction Registry, the pregnancy rate per transfer using IVF, ICSI is 25%, 30% respectively. Cryopreservation of sperm and embryos is a basic part of assisted reproduction, the result with oocytes freezing are still not satisfactory. Preimplantation genetic diagnosis is performed in few countries only due to technical barriers and ethical restrictions. Other methods like cloning, nuclear transfer, embryotherapy or gene manipulation using embryonic stem cells are still only subjects of research. CONCLUSION: During the last ten years, assisted reproduction showed rapid development in a number of treated couples and the spectrum of used methods changed radically.
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Técnicas Reproductivas , República Checa , Femenino , Humanos , Masculino , EmbarazoRESUMEN
Fully grown rabbit oocytes, isolated from preovulatory follicles, exhibit highly condensed bivalents within an intact germinal vesicle while a very low level of histone H1 kinase activity could be detected in their extracts. Chromatin condensation started in growing oocytes isolated from antral follicles presenting a diameter of 0.5 mm. This event was accompanied by a transient rise in histone H1 kinase activity which culminated in large antral follicles measuring 0.75 to 1 mm in diameter. However, the extent of histone H1 kinase activity observed in these growing oocytes remained far less important than that recorded in extracts prepared from in vitro cultured metaphase I and metaphase II oocytes. Moreover, this activity was insufficient to induce germinal vesicle breakdown which will only occur with an increasing efficiency, following in vitro culture of medium, large, and fully grown antral follicles.
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Cromatina/ultraestructura , Oocitos/enzimología , Oocitos/ultraestructura , Protamina Quinasa/metabolismo , Animales , Activación Enzimática , Femenino , Meiosis/fisiología , Oocitos/crecimiento & desarrollo , ConejosRESUMEN
Ultrastructural changes of the germinal vesicle during the growth of rabbit oocytes were studied by means of light and electron microscopy, 3H-uridine autoradiography, Ag-NOR staining and E-PTA staining. Particular interest was paid to the nucleologenesis and condensation of chromatin. In contrast to other mammalian species, chromosome condensation in rabbit oocytes occurred concomitantly with rRNA synthesis-dependent nucleolar compaction and preceded nuclear envelope breakdown and resumption of meiosis.