Live birth of monochorionic triamniotic triplets after in vitro fertilization and blastocyst transfer: case report and review of the literature

Monozygotic triplet pregnancies are very rare, even after in vitro fertilization (IVF). We present a case of a live birth of triplets from a monochorionic triamniotic pregnancy after blastocyst transfer. A 29-year-old woman underwent intracytoplasmic sperm injection (IVF-ICSI). Her medical history included a 4-year infertility treatment with ovarian stimulation, three cycles of intrauterine insemination, hysteroscopy and an unsuccessful attempt at IVF. During the second IVF attempt, the patient underwent ovulation simulation according to the long GnRH agonist protocol. Eleven metaphase II (MII) oocytes were injected with spermatozoa, resulting in four expanded blastocysts. In the end, two blastocysts (4AA) were transferred into the uterine cavity. Ultrasound examination performed at 7 weeks of gestation showed an ongoing triamniotic triplet pregnancy. Regular uterine contractions began at 33 weeks of gestation and a cesarean section was performed, resulting in a birth of three healthy girls, weighing 2060g (Apgar 7), 1860g (Apgar 6), 2000g (Apgar 6). After 13 days of hospitalization the infants and the mother were discharged home. Conclusion: The causes of monozygotic multiple gestations in spontaneous and ART pregnancies are poorly understood. They seem to occur more often after IVF. Any definitive relationship between particular stages of the IVF procedure and monozygotic multiple pregnancies remains controversial

Successful autotransplantation of cryopreserved ovarian tissue with recovery of the ovarian function

Cel pracy: Celem pracy był autologiczny przeszczep tkanki jajnikowej poddanej kriokonserwacji u pacjentki z przedwczesnym wygaśnięciem czynności jajników po agresywnej terapii onkologicznej. Materiał i metody: W 18 tygodniu ciąży u 28 letniej ciężarnej (C2P1) rozpoznano inwazyjną postać gruczołowego raka szyjki macicy. W 31 tygodniu przeprowadzono elektywne cięcie cesarskie, podczas, którego urodził się noworodek płci męskiej (1780g Apgar 8/10). Następnie wykonano radykalne wycięcie macicy techniką „nervesparing” z przydatkami, węzłami chłonnymi biodrowymi i około aortalnymi dolnymi. Jajniki poddano kriokonserwacji techniką powolnego zamrażania. Pacjentka przeszła cykl radiochemioterapii z następczą brachyterapią. Przedwczesne wygaśnięcie czynności jajników potwierdzono pomiarem hormonów płciowych: estradiol – 2 pg/ml, FSH – 96,52 IU/ml, LH – 37,55 IU/ml, AMH – 0,03 ng/ml. Trzynaście miesięcy po zabiegu chirurgicznym w powłokach brzusznych laparoskopowo wytworzono zachyłek otrzewnowy, w którym umieszczono rozmrożoną tkankę jajnikową Wyniki: Dziewięć tygodni po transplantacji uzyskano remisje objawów wypadowych, wzrost stężenia estradiolu (53 pg/ml), spadek FSH (64,89 IU/ml) i LH (33,39 IU/ml). Dwadzieścia cztery tygodnie po zabiegu zaobserwowano wysokie stężenia estradiolu (269 pg/ml), fizjologiczne stężenia FSH (5,92 IU/ml), LH (4,09 IU/ml) oraz wzrost stężenia AMH (0,37 ng/ml). Podczas badania ultrasonograficznego w przeszczepionej tkance jajnika uwidoczniono pęcherzyk dominujący. Wnioski: Przeszczep tkanki jajnikowej spowodował powrót czynność hormonalnej jajnika. Transplantacja tkanki jajnikowej może posłużyć, jako alternatywne leczenie przedwczesnej menopauzy spowodowanej agresywnym leczeniem onkologicznym.  Objectives: The aim of the study was autotransplantation of cryopreserved ovarian tissue to a patient suffering from premature ovarian failure caused by aggressive oncological therapy. Material and methods: A 28-year-old woman, GII PI, was diagnosed with invasive adenocarcinoma of the cervix at 18 weeks of gestation. At 31 weeks of gestation, a cesarean section was performed, resulting in the delivery of a healthy male newborn, followed by simultaneous, radical hysterectomy with bilateral salpingo-oophorectomy and lymphadenectomy. Half of each ovary was cryopreserved. The patient was scheduled for radiochemotherapy, supplemented with brachytherapy. After the intervention, the patient experienced menopausal symptoms. The basal hormonal levels were: estradiol – 2 pg/ml, FSH – 96.52 IU/ml, LH – 37.55 IU/ml, AMH – 0.03 ng/ml. Thirteen months after surgery, the peritoneal pocket was formed on the anterior abdominal wall during laparoscopy and heterotrophic autotransplantation of the frozen-thawed ovarian tissue was performed, replacing 59% of the tissue. Results: Nine weeks after transplantation, symptom resolution, an increase in estradiol (53 pg/ml), and a decrease in FSH (64.89 IU/ml) and LH (33.39 IU/ml) levels were noted. Twenty-four weeks after transplantation, high estradiol levels (269 pg/ml), normal level of FSH (5.92 IU/ml) and LH (4.09 IU/ml), and an increase in AMH (0.37 ng/ml) were observed. Follicular development in the transplanted ovarian tissue was confirmed. Conclusions: Cryopreservation and transplantation of ovarian tissue allowed to restore the ovarian function. It could offer an alternative physiological solution to treating premature ovarian failure caused by oncological therapy.

Detection of intracavitary lesions in 820 infertile women: comparison of outpatient hysteroscopy with histopathological examination

Objectives: The aim of the study was to compare the results of a diagnostic hysteroscopy with a histopathology examination (referential test) in a group of infertile women. Materials and methods: Eight hundred and twenty infertile patients were included in the study. The subjects with intracavitary lesions underwent operative hysteroscopy to enable the removal of polyps and intracavitary myomas. Endometrial biopsy was performed in all patients with no pathologies in hysteroscopy. The removed tissue underwent histopathological examination. Results: The mean age was 32.9 ±4.1. A total of 648 (79%) patients were diagnosed with primary and 172 (21%) with secondary infertility; 542 (66.1%) hysteroscopies were performed with no anesthesia and 278 (33.9%) hysteroscopies were performed in short total intravenous anesthesia. Sensitivity and specificity, accuracy, error, positive predictive value (PPV) and negative predictive value (NPV) of hysteroscopy in detecting endometrial lesions were 99.6%, 96.6%, 97.4%, 2.6%, 92.2% (PPV) and 99.8% (NPV), respectively. The agreement between hysteroscopy and pathology report was very high (kappa K=0.94). In case of normal uterine cavity, 562 of the 563 endometrial samples showed evidence of normal endometrium. In all 32 cases of resected submucosal myomas histopathology confirmed the hysteroscopic findings (sensitivity100%, specificity 100%, accuracy 100%, error 0%, kappa K=1.0). Both, hysteroscopy and histopathology confirmed the presence of endometrial polyps in 199 cases. The diagnosis of a polyp was not confirmed in histopathological findings (false-positive results) in 20 hysteroscopies. No endometrial polyps were missed during hysteroscopy. Sensitivity, specificity, accuracy, error, positive and negative predictive values in detecting endometrial polyps were 100%, 96.8%, 97.6%, 2.4%, 90.9% (PPV) and 100% (NPV), respectively. The Kappa coefficient agreement between hysteroscopy and histopathology for endometrial polyps was 0.91. Conclusions: Hysteroscopy is a method of high sensitivity and specificity in detecting pathologies, but in case of a pathology it cannot replace histological examination. Our result show that a routine endometrial biopsy performer in infertile patients with no pathology in hysteroscopy should not be recommended

Pro- and anti-inflammatory cytokines and growth factors in patients undergoing in vitro fertilization procedure treated with prednisone

Embryo implantation is a key moment in pregnancy. Abnormal production of pro- and anti-inflammatory cytokines, their receptors and other immune factors may result in embryo implantation failure and pregnancy loss. The aim of this study was to determine the profile of selected pro- and anti-inflammatory factors in the blood plasma of patients undergoing in vitro fertilization (IVF) and control women who achieved pregnancy after natural conception. The examined patients were administered steroid prednisone. We present results concern the plasma levels of IFN-ɣ, BDNF, LIF, VEGF-A, sTNFR1 and IL-10. We found that IVF patients receiving steroids differed significantly from patients who were not administered such treatment in terms of IFN-γ and IL-10 levels. Moreover, IVF patients differed in secretion of all tested factors with the fertile controls. Our results indicated that women who secrete at least 1409 pg/ml of sTNFR1 have a chance to become pregnant naturally and give birth to a child, while patients after IVF must achieve a concentration of 962.3 pg/ml sTNFR1 in blood plasma for successful pregnancy. In addition, IVF patients secreting VEGF-A above 43.28 pg/ml have a greater risk of miscarriage or a failed transfer in comparison to women secreting below this value. In conclusion, fertile women present a different profile of pro- and anti-inflammatory cytokines, and growth factors compared to patients with recurrent implantation failure (RIF)

Alternative reproductive tactics and sex-biased gene expression : the study of the bulb mite transcriptome

The sexes experience different selective pressures, which can lead to highly divergent phenotypes that are achieved via sex-biased gene expression. The effect of sexual dimorphism on the degree of sex-bias in gene expression can be studied in species characterized by sexually selected alternative male phenotypes. We analyzed gene expression in the bulb mite Rhizoglyphus robini (Acari, Acaridae), in which more sexually dimorphic, aggressive fighter males, possessing thickened legs of the third pair which are used to kill rivals, coexist with unarmored scrambler males. We sequenced transcriptomes of adult females and both types of males and de-novo assembled 114,456 transcriptome-based gene models (TGMs). Significantly more TGMs had male-biased expression than female-biased expression. Among TGMs that were over expressed in one, but not both, male morphs (compared to expression in females), we found about four times more fighter-biased genes than scrambler-biased genes. This demonstrates that the degree of expression bias reflects the degree of sexually selected dimorphism. However, the number of sex-biased genes was much higher than the number of genes differentially expressed between male morphs, and most male-biased genes were shared between morphs, suggesting that selection pressures act similarly on males irrespective of their morph. Furthermore, we found that male-biased genes evolved at a faster rate than female-biased genes, as evidenced by a higher rate of both gene-turnover and amino acid substitution, indicating that sexual selection, acting more strongly on males, accelerates the rate of molecular evolution. Interestingly, gene turnover was relatively higher, but amino acid substitution rate relatively lower among fighter-biased genes, suggesting that different components of sexual selection may have different effects on the evolution of sex-biased genes

Żywa ciąża bliźniacza otrzymana za pomocą metody ciągłego monitorowania zarodków u pacjentki z niepowodzeniami IVF-ET – opis przypadku i przegląd piśmiennictwa

Introduction: A standard assessment of embryo morphology at given time points does not always allow to transfer the embryo with the highest implantation potential. The effect of transfer of an improper embryo results in a lack of pregnancy or a miscarriage and, as a consequence, exposes the patient to unnecessary emotional stress and necessity to perform yet another transfer of frozen embryos. We present a case of a patient with earlier IVF failures. The use of time-lapse technique in this case helped to choose two good embryos. The transfer resulted in ongoing twin pregnancy. Material and methods: A 35-year-old woman with history of IVF-ET treatment failure was deemed eligible for an ICSI procedure because of the male factor. Ovarian stimulation was performed according to the agonist long protocol. Eight MII oocytes were fertilized and seven embryos were obtained. Continuous embryo monitoring was performed with the use of Primo Vision system. Forty-four hours after fertilization only 2 correctly developing embryos were identified. They were transferred on day 3. The development of the remaining 5 embryos was arrested. These embryos did not achieve the blastocyst stage on day 5-6 after fertilization. Forty days after embryo transfer, a twin pregnancy, confirmed with fetal heart rate of both fetuses, was revealed on ultrasound examination. Currently, the patient is at 27 weeks of ongoing twin gestation. Conclusions: The system of continuous embryo monitoring introduces new criteria for the examination of embryo development. These new parameters can be useful in clinical practice. However, prospective randomized studies are necessary to provide data confirming the usefulness of time-lapse technique in IVF treatment.Wstęp: Standardowa obserwacja zarodków w procedurze IVF nie zawsze pozwala na wybór do transferu zarodka o najwyższym potencjale implantacyjnym. Naraża to pacjentkę na stres emocjonalny spowodowany brakiem ciąży, ewentualnym poronieniem oraz koniecznością powtórnego transferu zarodków mrożonych. W pracy przedstawiamy przypadek pacjentki po dwóch nieudanych próbach zapłodnienia pozaustrojowego in vitro, u której podczas kolejnego IVF-ICSI system time-lapse pomógł w wyborze dwóch prawidłowych zarodków. Zarodki przetransferowano w 3 dobie obserwacji i otrzymano żywą prawidłowo przebiegającą ciążę bliźniaczą. Materiał i metody: Trzydziestopięcioletnia pacjentka z dwoma niepowodzeniami IVF-ICSI została zakwalifikowana do ponownej procedury IVF-ICSI. Wskazaniem do leczenia był czynnik męski. Po stymulacji jajników według protokołu długiego uzyskano 8 prawidłowych komórek MII, uzyskując 7 zarodków. Do ciągłego monitorowania rozwoju zarodków wykorzystano system Primo Vision (Cryo Innovation). 44 godziny po ICSI zidentyfikowano tylko dwa prawidłowo rozwijające się zarodki, które podano do jamy macicy w trzeciej dobie obserwacji. Pozostałe embriony nie wykazały cech dalszych podziałów. Czterdzieści dni po transferze w badaniu ultrasonograficznym uwidoczniono ciążę bliźniaczą dwukosmówkową z czynnością serca obydwu płodów. Obecnie pacjentka jest w 27 tygodniu prawidłowo rozwijającej się ciąży bliźniaczej. Wnioski: System ciągłego monitorowania rozwoju zarodków wprowadza nowe kryteria ich oceny, które mogą być przydatne w praktyce klinicznej. Konieczne są jednak prospektywne randomizowane badania oceniające wartość systemów time-lapse w leczeniu metodą zapłodnienia pozaustrojowego

Wpływ energii wiązek fotonowych na rozkład dawek dla planów IMRT i VMAT

Purpose. Estimation of the difference between photon X-6MV and X-20MV beams for IMRT and VMAT plan’s, in respect to dose-volume results and irradiation time (MU).Materials and methods. For each of 74 selected patients four plans, two IMRT with X-6MV & X-20MV beams and two VMAT: X-6MV & X-20MV arcs, were performed. Patients were divided into two groups according to tumor localization: head & neck (H&N) and pelvis. Those localizations were chosen to highlight potential differences regarding the depth of target volume. Each plan was optimized using the same plan objectives and constraints. Plans were compared according to dose-volume results for target and Organs at Risk (OaR’s) with Radiation Planning Index (RPI), and beamon time regarding the monitor units (MUs).Results. The mean RPI factor for both technique (IMRT/VMAT) and energies (X-6MV/X-20MV) were similar for H&Nregion in the range of 0.2310–0.2934 and for the pelvis region the range was 0.3683–0.4007. The difference were not statistically significant (p > 0.05), showing the photon between 6–20 MV, doesn’t influence the dose-volume results, for both localization: H&N and pelvis. The mean monitor units in IMRT plans varied from 765 MU to 1116 MU, as inVMAT plans it was from 325 MU to 492 MU. Generally, the number of MU on IMRT technique is greater than MU’s inVMAT (difference statistically significant), regardless of the beam energy (X-6MV, X-20MV) and localization (H&N, pelvis). Both techniques; IMRT and VMAT plans with higher photon energy, showed shorter irradiation time (expressedin MU). But, only for pelvic region on VMAT technique, is statistically significant (p = 0.0467).Conclusions. On average, photons beam, between 6–20 MV don’t induce significant dose-volume difference. However, higher energy used for planning regions other than head & neck, minimizes the number of MUs and significantly reduces the time of irradiation. Furthermore, reduction of beam on time can be achieved by using VMAT plan rather than IMRT plan.Cel. Ocena różnic w planach IMRT i VMAT pomiędzy wiązkami X-6MV i X-20MV w odniesieniu do uzyskanych rozkładów dawek oraz czasu napromieniania (MU).Materiały i metody. Na potrzeby porównania wybrano 74 pacjentów, następnie dla każdego obliczono 4 plany: dwa w technice IMRT z wiązkami X-6MV i X-20MV oraz dwa w technice VMAT z łukami X-6MV i X-20MV, uzyskując rozkłady dawek. Pacjentów podzielono na dwie grupy zgodnie z obszarem napromienienia: rejon głowy/szyi oraz rejon miednicy. Lokalizacje zostały wybrane w celu uwidocznienia ewentualnych różnic wynikających z różnych głębokości na których zdefiniowano obszar tarczowy. Każdy plan był optymalizowany z wykorzystaniem z tych samych wytycznych i ograniczeń związanych z obszarami anatomicznymi. Plany zostały następnie porównane pod względem dawek dla obszarów tarczowych i organów krytycznych z użyciem współczynnika RPI (Radiation Planning Index). Następnie porównano czas napromieniania w ujęciu liczby jednostek monitorowych MU.Wyniki. Średnie współczynniki RPI dla obydwóch technik (IMRT/VMAT) i energii wiązek/łuków (X-6MV/X-20MV) były porównywalne i zawierały się w przedziale 0,2310–0,2934 dla rejonu głowy i szyi oraz 0,3683–0,4007 dla rejonu miednicy. Różnice nie były statystycznie istotne (p > 0,05), wykazując brak wpływu wyboru energii wiązek/łuków na uzyskane rozkłady dawek dla obydwu obszarów napromieniania: głowy/szyi oraz miednicy. Średnia liczba jednostek monitorowych zawierała się w przedziale 765 MU do 1116 MU dla planów w technice IMRT oraz 325 MU do 492 MU dla planów w technice VMAT. Liczba jednostek monitorowych w planach wykorzystujących technikę IMRT była zawsze większa od liczby jednostek w planach z techniką VMAT (potwierdzona istotnością statystyczną), niezależnie od stosowanej energii wiązek/łuków oraz napromienianego regionu głowa/szyja/miednica. Wykorzystanie wiązek/łuków o wyższej energii podczas planowania z wykorzystaniem każdej techniki (IMRT i VMAT) skutkowało zmniejszeniem czasu napromieniania (rozpatrywanego w oparciu o jednostki monitorowe), jednak tylko w przypadku techniki VMAT i obszaru miednicy jest to poparte istotnością statystyczną (p = 0,0467).Wnioski. Zasadniczo wybór energii wiązek z przedziału 6–20 MV nie wprowadza znaczących różnic w uzyskiwanych rozkładach dawek, jednakże wykorzystanie wyższej energii w obszarach napromieniania innych niż głowa/szyja zmniejsza liczbę jednostek monitorowych i znacząco skraca czas napromieniania. Ponadto dalsze skrócenie czasu napromieniania jest możliwe z zastosowaniem techniki VMAT zamiast IMRT

No evidence for reproductive isolation through sexual conflict in the bulb mite Rhizoglyphus robini

Sexual conflict leading to sexual antagonistic coevolution has been hypothesized to drive reproductive isolation in allopatric populations and hence lead to speciation. However, the generality of this speciation mechanism is under debate. We used experimental evolution in the bulb mite Rhizoglyphus robini to investigate whether sexual conflict promotes reproductive isolation measured comprehensively to include all possible pre- and post-zygotic mechanisms. We established replicate populations in which we either enforced monogamy, and hence removed sexual conflict by making male and female evolutionary interests congruent, or allowed promiscuity. After 35 and 45 generations of experimental evolution, we found no evidence of reproductive isolation between the populations in any of the mating systems. Our results indicate that sexual conflict does not necessarily drive fast reproductive isolation and it may not be a ubiquitous mechanism leading to speciation

454 sequencing reveals extreme complexity of the class II Major Histocompatibility Complex in the collared flycatcher

<p>Abstract</p> <p>Background</p> <p>Because of their functional significance, the Major Histocompatibility Complex (MHC) class I and II genes have been the subject of continuous interest in the fields of ecology, evolution and conservation. In some vertebrate groups MHC consists of multiple loci with similar alleles; therefore, the multiple loci must be genotyped simultaneously. In such complex systems, understanding of the evolutionary patterns and their causes has been limited due to challenges posed by genotyping.</p> <p>Results</p> <p>Here we used 454 amplicon sequencing to characterize MHC class IIB exon 2 variation in the collared flycatcher, an important organism in evolutionary and immuno-ecological studies. On the basis of over 152,000 sequencing reads we identified 194 putative alleles in 237 individuals. We found an extreme complexity of the MHC class IIB in the collared flycatchers, with our estimates pointing to the presence of at least nine expressed loci and a large, though difficult to estimate precisely, number of pseudogene loci. Many similar alleles occurred in the pseudogenes indicating either a series of recent duplications or extensive concerted evolution. The expressed alleles showed unambiguous signals of historical selection and the occurrence of apparent interlocus exchange of alleles. Placing the collared flycatcher's MHC sequences in the context of passerine diversity revealed transspecific MHC class II evolution within the Muscicapidae family.</p> <p>Conclusions</p> <p>454 amplicon sequencing is an effective tool for advancing our understanding of the MHC class II structure and evolutionary patterns in Passeriformes. We found a highly dynamic pattern of evolution of MHC class IIB genes with strong signals of selection and pronounced sequence divergence in expressed genes, in contrast to the apparent sequence homogenization in pseudogenes. We show that next generation sequencing offers a universal, affordable method for the characterization and, in perspective, genotyping of MHC systems of virtually any complexity.</p