315 IVF-DERIVED CROSSBRED EMBRYOS PRODUCED WITH SEXED SEMEN AND TRANSFERRED IN PAIRS TO BOS TAURUS × BOS INDICUS COWS AND HEIFERS

2015 ◽  
Vol 27 (1) ◽  
pp. 246
Author(s):  
S. Romo ◽  
S. Castañeda ◽  
C. A. Hernandez ◽  
J. H. Mendoza ◽  
F. J. Trejo ◽  
...  
Keyword(s):  
Production Efficiency ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Follicular Aspiration ◽  
Fisher’S Exact Test ◽  
Sexed Semen ◽  
Rectal Palpation

Biotechnology has continued to evolve rapidly, allowing the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include the in vitro maturation (IVM) and IVF of oocytes, sperm sexing and embryo transfer (ET) to recipient females to obtain pregnancies and offspring. These modern assisted-reproduction techniques (ART) can help produce twin pregnancies and calving of a pre-determined sex. The aim of this study was to produce a high proportion of female bovine embryos in vitro using X-chromosome-selected sexed semen and to transfer them in pairs to recipient females, in order to evaluate the efficiency of transferring two female embryos in both cows and heifers. Cebu-cross ovaries were obtained from a local slaughterhouse and transported to a nearby laboratory in Chiapas, Mexico, to obtain cumulus-oocyte complexes by follicular aspiration and culture in maturation medium for 24 h. For IVF, frozen X-sorted semen (Milking Gyr and Holstein breeds, 90% purity, Sexing Technologies, Navasota, TX, USA) was used. Gametes were co-incubated for 22 h, then moved to embryo development medium and cultured for 7 days. Recipient Cebu-cross commercial cows (n = 98) and heifers (n = 50) were synchronized, using intravaginal devices impregnated with progesterone, administering eCG and prostaglandin at withdrawal. Seven days after heat, 88 recipients were subjected to non-surgical ET (59 cows and 29 heifers). Embryo transfers were performed in Tamaulipas and Veracruz, Mexico, and were divided into 2 groups: A) cows, and B) heifers. Only grade-1 embryos were selected for ET. Two embryos were loaded in a single 0.25 mL French straw and transferred to the uterine horn ipsilateral to the ovary with a corpus luteum. Pregnancy diagnosis was performed by ultrasound or rectal palpation 60 days after ET. A Fisher's exact test (SPSS v. 16.0, SPSS Inc., Chicago, IL, USA) was used to determine statistical differences (α = 0.05). Of IVF oocytes, 176/180 (98%) and 242/300 (81%) were fertilized, producing 96/180 (53.3%) Milking Gyr (semen)-Cebu (oocytes) and 92/300 (30.7%) Holstein (semen)-Cebu (oocytes) grade-1 embryos, respectively. Of the 88 recipients, 33 were pregnant (37.5%), however, it was not possible (at that time) to determine the number and sex of fetuses developing in utero. Overall, 8 of the 29 heifers were pregnant (27.6%), compared to 25 pregnancies in 59 cows (42.4%). For heifers, the pregnancy results for transferring Milking Gyr-Cebu embryos (4/11) versus Holstein-Cebu embryos (4/18) were not significantly different (P = 0.433, two-tailed Fisher's exact test). A similar comparison could not be made for cows given smaller sample sizes due to the extra variable of having taken place on several different ranches (n = 11). The concept herein is that smaller twin female crossbred calves may reduce issues associated with freemartinism and dystocia while still maintaining the vigor of crossbred offspring. Results from this research can help contribute to the study and development of ART for increasing cattle production efficiency.

169 OVUM PICK-UP, IN VITRO EMBRYO PRODUCTION, AND EMBRYO TRANSFER IN CATTLE IN TROPICAL AND SUBTROPICAL MEXICO

2016 ◽  
Vol 28 (2) ◽  
pp. 215
Author(s):  
S. Castañeda ◽  
S. Romo ◽  
M. E. Kjelland
Keyword(s):  
Embryo Transfer ◽  
Production Efficiency ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Hormonal Treatment ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Sexed Semen ◽  
In Vitro Embryo Production

Biotechnology continues to evolve rapidly, allowing the development of artificial reproductive techniques (ART) to increase reproductive efficiency and contribute to the genetic improvement of domestic animals. The present study examines the results obtained after 30 months of starting a commercial practice for ovum pickup (OPU) in vitro embryo production (IVP) and embryo transfer (ET) in cattle in tropical and subtropical Mexico. This research was conducted from 2013–2015 in beef and dairy cattle kept under different environmental and management conditions in 6 states (Chiapas, Oaxaca, Tabasco, Tamaulipas, Veracruz, and Yucatan). Oocytes were collected by OPU, without hormonal treatment, from 10 donor cows: 2 Bos taurus (Bt), 5 Bos indicus (Bi), and 3 Bt × Bi. A total of 98 oocyte recovery sessions were performed on 756 cows and produced 12 524 viable oocytes (1349 GI, 3383 GII, 7792 GIII), which were sent to a central laboratory for IVP. Both conventional and sexed semen were used for IVF, from 9 breeds: 2 Bt, 5 Bi and 2 Bt × Bi. The overall cleavage rate was 69% (8587/12 524). The embryo production efficiency rate was 31% (3905/12 524). Fresh sexed and conventional embryos were transferred to recipients synchronised with the following protocol: Day 0, application of an intravaginal device (ID) with progesterone and 2 mg of oestradiol benzoate IM; Day 8, removal of the ID, 400 IU eCG IM, 0.5 mg cloprostenol sodium IM, and 0.5 mg oestradiol cipionate IM. Day 10 was considered the day of oestrus. Pregnancy rate after ET was 38% (945/3905). The average number of viable oocytes per donor cow was 16.57; the average number of transferred embryos per donor cow was 5.17, and the average number of pregnancies per donor cow was 1.25. The OPU-IVP were successful in producing pregnancies even under several adverse conditions, such as a tropical environment, many donors being prepuberal females and the majority of the adult cows having a previous non-productive history due to hormonal mishandling in superovulation programs or because of overfeeding for show purposes. The consequences of these factors can be observed in the lower overall cleavage rate obtained (69%), compared to the expected (75%). Some variables require further analysis (e.g. different OPU and ET technicians, time of year, cow age, cow breed, and use of conventional and sexed semen). A successful IVP practice has to face and overcome multiple problems that may arise in some geographic locations but, perhaps, not in others. It is interesting to note the use of sexed versus conventional embryos, of the 98 ET sessions, 25 involved sexed embryos versus 73 for conventional embryos. The use of sexed semen for OPU-IVP and ET in this region of Mexico continues to expand. We thank Genemex Internacional and the ranch owners that were involved with this research.

147 AGGREGATION OF IN VIVO FERTILIZED AND PARTHENOGENETICALLY ACTIVATED MOUSE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 221
Author(s):  
H. Fernandes ◽  
P. T. Mihara ◽  
B. Cazari ◽  
I. P. Emanuelli ◽  
M. F. G. Nogueira
Keyword(s):  
Cell Mass ◽  
Blastocyst Stage ◽  
Strontium Chloride ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Fisher’S Exact Test ◽  
Inverted Microscope ◽  
Parthenogenetic Embryos

Embryonic chimerism – mixing of cells originated from at least two different fertilizations – has been used as a tool for stem cell pluripotency diagnosis, transgenic rodent production, and organogenesis studies. Additionally, parthenotes are used for studies related to gene imprinting and the ability of their cells to compose the placenta and/or an adult animal. The aim of this work was to validate the production and characterize developmental kinetic of parthenogenetic embryos, obtained from C57BL/6 EGFP mice (EGFP), to determine their potential to produce chimeras, and to localize parthenogenetic cells on the produced blastocyst. Embryos were harvested from superovulated females. For the aggregation, pre-compaction IVF (Swiss Webster/SW strain) or parthenogenetic activated (PGA; SW and EGFP) embryos were used. For PGA, strontium chloride hydrate (5 mM for 6 h) was used. Two experiments were outlined in order to: i) evaluate the development and kinetic from IVF (IVF and in vitro developed; 2-cell stage embryos, n = 53) and from PGA (parthenogenetically activated and in vitro developed; n = 409 oocytes) techniques, both from SW; ii) evaluate the aggregation between pairs of control embryos (C; n = 20, 4-cell in vivo produced embryos from SW) or pairs of 4-cell in vivo produced SW embryos and 4- to 8-cell PGA EGFP embryos (parthenogenetic embryo, PG; n = 40). After manipulation (removal of the zona pellucida and approximation of pairs, for C and PG groups), all the groups were kept in vitro culture (37°C, 5% CO2, and saturated humidity) for 48 to 60 h (C and PG), or up to blastocyst stage (IVF and PGA). Chimerism rate and parthenogenetic cells fluorescence (pre- and post-culture) were evaluated under an inverted microscope with epifluorescence source, and digital images were captured (Eclipse Ti and NIS-Elements, Nikon, Japan, respectively), by merging visible and UV light images. The rate of PGA (EGFP oocytes) was 54.5% (66/121), assessed 6 h post-activation by the presence of at least one pronucleus. The assessment was performed with 20× magnification and Hoffman modulation contrast in an inverted microscope. The rate of blastocyst production between IVF and PGA was significantly different (71.4 and 12.9%, respectively; P < 0.001, Fisher’s exact test). When developmental kinetic was evaluated there a difference in the average time to the majority of embryos reach blastocyst stage for IVF (48 h) and PGA (120 h) was observed. The chimerism rate was assessed by the presence of a single and cohesive cell mass (C) or by the incorporation of EGFP cells in the SW embryo (PG). There was difference (P = 0.006; Fisher’s exact test) between C (55.0%; 11/20) and PG (17.5%; 7/40) chimerism rates, assessed from 48 to 60 h of culture. In PG group, the incorporation of EGFP cells in the obtained chimeras was mainly detected in the trophectoderm although one chimeric blastocyst contained EGFP cells only in the inner cell mass. It was concluded that parthenogenetic embryos have a slower developmental kinetic (until 120 h of culture) than embryos derived from IVF. Acknowledgment to FAPESP for fellowship and funding.

159 USE OF PHYTOHEMAGGLUTININ-L, AS AN AGGLUTINATOR AGENT, TO PRODUCE CHIMERAS OF IVF BOVINE EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 238
Author(s):  
I. P. Emanuelli ◽  
B. F. Agostinho ◽  
M. P. M. Mancini ◽  
C. M. Barros ◽  
M. F. G. Nogueira
Keyword(s):  
Embryonic Stem ◽  
Blastocyst Stage ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Stage Of Development ◽  
Fisher’S Exact Test

Embryonic chimeras have been used as a tool to understand embryogenesis and organogenesis, as well as to prove, in vivo, the pluripotency of the embryonic stem cells. One of the techniques used to obtain embryonic chimeras is aggregation, which can be performed with intact or half-embryos and in different stages of the development, produced by in vivo or in vitro systems and in different wells. However, its efficiency tends to reduce when advanced stages, such as morulae and blastocysts, are used. The aim of this work was to evaluate the effect of the treatment with an agglutinating agent (phytohemagglutinin-L; PHA) in the percentage of chimeras produced with IVF bovine embryos. Bovine ovaries (from abattoir) were used to obtain 270 COC that were matured in drops (90 μL) of TCM-199 bicarbonate medium, supplemented with 10% of FCS, and incubated in vitro for 22 to 24 h. The fertilization occurred in TALP-IVF medium, and the COC were maintained in the incubator for 18 h. After fertilization, the presumptive zygotes were transferred to SOF culture medium to in vitro culture. In vitro maturation, fertilization, and culture were performed under 38.5°C, 5% CO2 in air and saturated humidity. The chimerism by aggregation was tested between 2 intact (zona-free) 8- to 16-cell stage embryos in the presence (G1, n = 16) or absence of PHA (G2, n = 14) and between one half-morula and one half-blastocyst with (G3, n = 15) or without PHA (G4, n = 12). The embryos in groups G1 and G3 were treated with PHA in a concentration of 500 μLg mL-1 for 3 min. After PHA treatment, the pairs of embryos were allocated in wells, under previously described culture conditions, until expanded blastocyst stage could be observed (Day 7 of culture). At 24 h of culture, embryonic aggregation pairs were first evaluated to detect only cohesive masses of cells. The results (chimerism rate) were 62.5%, 42.9%, 40.0%, and 25.0%, respectively, for groups G1, G2, G3, and G4. There were no significant differences neither among groups (chi-square, P = 0.252) nor between G1 and G2 (P = 0.464), G3, and G4 (P = 0.683; Fisher’s exact test). Main effects as use of PHA (G1 + G3 v. G2 + G4, P = 0.284) and stage of embryos (G1 + G2 v. G3 + G4, P = 0.183; Fisher’s exact test) were not statistically significant. However, when all groups were compared, the power of the performed test (0.354) was below the desired power of 0.800 (i.e. one must be cautious in over-interpreting the lack of difference among them). In the conditions of this study, it was concluded that the treatment with PHA did not increase the rate of aggregation in the embryonic chimera production, even for half-embryos in advanced stage of development (morulae and blastocysts). Granted by FAPESP, Brazil: 06/06491-2 and 07/07705-9 (MFGN) and 07/04291-9 (MPMM).

scholarly journals Effect of the antral follicle count of Bos taurus × Bos indicus dairy cows on in vitro embryo production

2020 ◽  
pp. 2171-2178
Author(s):  
Sheila Merlo Garcia ◽  
Paula Alvares Lunardelli ◽  
Kleber Luciano Ancioto ◽  
Eduardo Cardoso de Oliveira ◽  
Larissa Zamparone Bergamo ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Antral Follicle ◽  
Estradiol Benzoate ◽  
Antral Follicle Count ◽  
Success Rates ◽  
In Vitro Production ◽  
Follicular Aspiration ◽  
Vitro Production

This study aimed to evaluate the potential of Girolando (Bos taurus × Bos indicus) cows with high and low antral follicle counts (AFC) for the in vitro production of bovine embryos (IVEP), as well as the pregnancy rates of the recipients of these embryos. Girolando cows (Bos taurus × Bos indicus) were classified as high and low AFC when they had 35-52 (n = 13) and 11-17 follicles (n = 15), respectively. All animals were subjected to repeated follicular aspiration [Ovum pick-up (OPU)] and subsequent IVEP sessions. The synchronization protocol of the recipients was performed on a random day of the estrous cycle (Day 0) with the implantation of progesterone, estradiol benzoate, and prostaglandin. The high AFC group had higher aspirated oocyte/OPU (42.6 ± 5.2 vs. 14.6 ± 1.9; p < 0.01) and cultured oocyte/OPU (38.1 ± 6.6 vs. 12.3 ± 2.8; p < 0.01) averages as well as a higher blastocyst percentage on D7 (23.0 ± 1.0% vs. 18.4 ± 1.5%; p < 0.05) and higher pregnancy rate (42.7 ± 2.7% vs. 39.7 ± 4.6%; p < 0.05) than the low AFC group. Thus, we can conclude that animals with high AFC had better IVEP success rates than animals with low AFC.

Effects of sexed semen and interactive effects on commercial in vitro embryo production when oocytes are collected from cows of Bos indicus, and Bos taurus breeding and crossbred cows of these subspecies

2015 ◽  
Vol 156 ◽  
pp. 58-63 ◽  
Author(s):  
William Omar Contreras Lopez ◽  
Hernando Raphael Alvis-Miranda ◽  
Andres Felipe Gamarra ◽  
Bibiana Rendon ◽  
Diego Armando Vega Borda ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Bos Indicus ◽  
Interactive Effects ◽  
Embryo Production ◽  
Crossbred Cows ◽  
Sexed Semen ◽  
In Vitro Embryo Production

Δράση του φυτού βαλσαμόχορτο (Hypericum perforatum) στον καρκίνο της ουροδόχου κύστης

2019 ◽  
Author(s):  
Ιωάννης Τσίμαρης
Keyword(s):  
Hypericum Perforatum ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Rp Hplc ◽  
Fisher’S Exact Test

Σκοπός της παρούσας διδακτορικής διατριβής ήταν η συστηματική και εις βάθος έρευνα της αντικαρκινικής δράσης του φυτού βαλσαμόχορτου γνωστού και ως Hypericim Perforatum (HP) κατά του καρκίνου της κύστης. Με την βοήθεια υγρής χρωματογραφίας υψηλής απόδοσης αντίστροφης φάσης (RP-HPLC) αναγνωρίστηκε ότι το κύριο συστατικό του υδατικού διαλύματος που λαμβάνεται στα πλαίσια της παραδοσιακής ηπειρώτικης ιατρικής είναι η υπερφορίνη (2,36 ± 0,03 mg/g).Τοπειραματικό μοντέλο περιλάμβανε αρσενικά ποντίκια BULBc τα οποία εκτέθηκαν σε νιτροζαμίνη (OH-BBN) σε συγκέντρωση 1000ppm μέρα παρά μέρα για 6 εβδομάδες. Θυσιάστηκαν μετά από 26 εβδομάδες. Τα ιστολογικά δεδομένα έδειξαν ότι το υδατικό διάλυμα ασκεί στατιστικώς σημαντική αντικαρκινική δράση (Fisher’s exact test p=0.048) όταν χορηγείται για 26 εβδομάδες αφού επιδράσει ο καρκινογόνος παράγοντας. Η χορήγηση του εκχυλίσματος το διάστημα που λάμβαναν νιτροζαμίνη δεν εμφάνισε σημαντική αντικαρκινική δράση (Fisher’s exact test p=0.093). Στην συνέχεια προχωρήσαμε στην αναζήτηση του πιο δραστικού παραγώγου του φυτού, είτε με μορφή διαλυμάτων, είτε με την μορφή εκχυλισμάτων, είτε μετά από κλασματοποίηση. Η ανάλυση των συστατικών τους έγινε με RP-HPLC. Η αντικαρκινική δράση αποτιμήθηκε σε καρκινικές κυτταρικέςσειρές ουροδόχου κύστεως Τ24 και NBT-II σε συνθήκες σκότους. Η διαδικασία κλασματοποίησης ανέδειξε το έκτο κατά σειρά κλασματοποίησης κλάσμα του Εκχυλίσματος Πετρελαϊκού Αιθέρα (ΕΠΑ6) ως το πλέον δραστικό (LC50 0.17 μg/mL). Στην αναζήτηση των συστατικών που ευθύνονται για την αντικαρκινική δράση διαπιστώθηκε ότι αυτή δεν οφείλεται στην υπερφορίνη αλλά σε άγνωστες ουσίες οι οποίες δεν ήταν δυνατόν να ταυτοποιηθούν στην δεδομένη χρονική στιγμή. Ηδοκιμασία TUNEL απέδειξε πως ο κυτταρικός θάνατος επέρχεται μέσω της απόπτωσης. Από την φυσικοχημική ανάλυση των εκχυλισμάτων διαπιστώθηκε πως το Πολικό Μεθανολικό Κλάσμα (ΠΜΚ), σε μορφή πούδρας, περιέχει υπερικίνη σε συγκέντρωση 0.57% μόνο, είναι υδατοδιαλυτό, σταθερό σε θερμοκρασία δωματίου καθιστώντας το ενδιαφέρον φωτοευαισθητοποιητή για περαιτέρω μελέτη.Προχωρήσαμε σε σύγκριση του ΠΜΚ, σε διάφορες συγκεντρώσεις με το Photofrin® , εγκεκριμένη από το FDA ουσία για την Φωτοδυναμική Θεραπεία (ΦΔΘ). Το ΠΜΚ στη συγκέντρωση των 60 μg/mL, προκαλούσε κυτταρικό θάνατο στο 86% και στο 80% των Τ24 και RT4 καρκινικών κυττάρων αντίστοιχα. Το Photofrin® εμφάνισε κυτταρικό θάνατο στο 77% των Τ24 μόνο. Μετά από σύγκριση πηγών φωτισμού λευκού φωτός και ΠΜΚ σε Τ24 κύτταρα διαπιστώθηκε ότι η πιο αποτελεσματική ήταν η πηγή ψυχρού φωτισμού με λάμπα αλογόνου (Storz 482 Cold Light Fountain). Πειραματιζόμενοι με θηλυκούς Wistar αρουραίους με ποικίλες ενδοκυστικές δόσεις ΠΜΚ, με κλιμακούμενους χρόνους ενδοκυστικής παραμονής και ποικίλες ενέργειες διέγερσης καταλήξαμε στην δοσολογία των 36.5mg/ml του ΠΜΚ, δεκαπεντάλεπτη ενδοκυστική παραμόνή και διέγερση με 700J/cm2 λευκού φωτός. Με βάση το παραπάνω προχωρήσαμε σε πειραματικό μοντέλο ορθότοπης εμφύτευσης ουροθηλιακών καρκινικών κυττάρων ΑΥ27. Είχαμε πλήρη (100%) εξάλειψη επιφανειακών καρκινικών βλαβών αλλά και του φυσιολογικού ουροθηλίου στις 30 ημέρες μετά την θεραπεία. Το ουροθήλιο αναγεννάτε πλήρως, χωρίς την ύπαρξη ουλών, 90 ημέρες μετά την φωτοδυναμική θεραπεία. Συμπερασματικά το εκχύλισμα του υπέρικου, στη δοσολογία που το λάμβαναν οι ασθενείς, παρουσιάζει στατιστικώς σημαντική αντικαρκινική δράση όταν χορηγείταιμετά την δράση του καρκινογόνου. Η αντικαρκινική δράση που παρατηρείται in vitro στο αποδίδεται σε άγνωστες, προς το παρόν ουσίες, που περιέχονται στο ΕΠΑ6. Το ΠΜΚ εμφανίζει σημαντική αντικαρκινική δράση τόσο in vitro, μετά από διέγερση με λέιζερ ή λευκό φως, όσο και in vivo, με πλήρη εξάλειψη των καρκινικών λαβών, χωρίς την εμφάνιση ουλών στην ουροδόχου κύστη με πλήρως αναγεννημένο του ουροθήλιο μετά από 90 ημέρες.

scholarly journals TERT promoter hotspot mutations and gene amplification in metaplastic breast cancer

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Edaise M. da Silva ◽  
Pier Selenica ◽  
Mahsa Vahdatinia ◽  
Fresia Pareja ◽  
Arnaud Da Cruz Paula ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Amplification ◽  
Genetic Alterations ◽  
Wild Type ◽  
Histologic Subtype ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Fisher’S Exact Test ◽  
Hotspot Mutations ◽  
Or Gene

AbstractMetaplastic breast cancers (MBCs) are characterized by complex genomes, which seem to vary according to their histologic subtype. TERT promoter hotspot mutations and gene amplification are rare in common forms of breast cancer, but present in a subset of phyllodes tumors. Here, we sought to determine the frequency of genetic alterations affecting TERT in a cohort of 60 MBCs with distinct predominant metaplastic components (squamous, 23%; spindle, 27%; osseous, 8%; chondroid, 42%), and to compare the repertoire of genetic alterations of MBCs according to the presence of TERT promoter hotspot mutations or gene amplification. Forty-four MBCs were subjected to: whole-exome sequencing (WES; n = 27) or targeted sequencing of 341-468 cancer-related genes (n = 17); 16 MBCs were subjected to Sanger sequencing of the TERT promoter, TP53 and selected exons of PIK3CA, HRAS, and BRAF. TERT promoter hotspot mutations (n = 9) and TERT gene amplification (n = 1) were found in 10 of the 60 MBCs analyzed, respectively. These TERT alterations were less frequently found in MBCs with predominant chondroid differentiation than in other MBC subtypes (p = 0.01, Fisher’s exact test) and were mutually exclusive with TP53 mutations (p < 0.001, CoMEt). In addition, a comparative analysis of the MBCs subjected to WES or targeted cancer gene sequencing (n = 44) revealed that MBCs harboring TERT promoter hotspot mutations or gene amplification (n = 6) more frequently harbored PIK3CA than TERT wild-type MBCs (n = 38; p = 0.001; Fisher’s exact test). In conclusion, TERT somatic genetic alterations are found in a subset of TP53 wild-type MBCs with squamous/spindle differentiation, highlighting the genetic diversity of these cancers.

scholarly journals Association of P10L Polymorphism in Melanopsin Gene with Chronic Insomnia in Mexicans

2021 ◽  
Vol 18 (2) ◽  
pp. 571
Author(s):  
Bianca Ethel Gutiérrez-Amavizca ◽  
Ernesto Prado Montes de Oca ◽  
Jaime Paul Gutiérrez-Amavizca ◽  
Oscar David Castro ◽  
Cesar Heriberto Ruíz-Marquez ◽  
...  
Keyword(s):  
Statistical Power ◽  
Medical Condition ◽  
Severity Index ◽  
Recessive Model ◽  
Chronic Insomnia ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Increased Risk ◽  
Fisher’S Exact Test ◽  
And Function

The aim of this pilot study was to determine the association of the P10L (rs2675703) polymorphism of the OPN4 gene with chronic insomnia in uncertain etiology in a Mexican population. A case control study was performed including 98 healthy subjects and 29 individuals with chronic insomnia not related to mental disorders, medical condition, medication or substance abuse. Samples were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Genetic analyses showed that the T allele of P10L increased risk to chronic insomnia in a dominant model (p = 1 ×10−4; odds ratio (OR) = 9.37, CI = 8.18–335.66, Kelsey statistical power (KSP) = 99.9%), and in a recessive model (p = 7.5 × 10−5, OR = 9.37, KSP = 99.3%, CI = 2.7–34.29). In the insomnia group, we did not find a correlation between genotypes and chronotype (p = 0.219 Fisher’s exact test), severity of chronic insomnia using ISI score (p = 0.082 Fisher’s exact test) and ESS score (p ˃ 0.999 Fisher’s exact test). However, evening chronotype was correlated to daytime sleepiness severity, individuals with an eveningness chronotype had more severe drowsiness according to their insomnia severity index (ISI) score (p = 0.021 Fisher’s exact test) and Epworth sleepiness scale (ESS) score (p = 0.015 Fisher’s exact test) than the morningness and intermediate chronotype. We demonstrated that the T allele of the P10L polymorphism in the OPN4 gene is associated with chronic insomnia in Mexicans. We suggest the need to conduct larger studies in different ethnic populations to test the probable association and function of P10L and other SNPs in the OPN4 gene and in the onset of chronic insomnia.

scholarly journals Detection of AZF microdeletions and reproductive hormonal profile analysis of infertile sudanese men pursuing assisted reproductive approaches

BMC Urology ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Hassan Osman Alhassan Elsaid ◽  
Tarteel Gadkareim ◽  
Tagwa Abobakr ◽  
Eiman Mubarak ◽  
Mehad A. Abdelrhem ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Klinefelter Syndrome ◽  
Semen Analysis ◽  
Control Group ◽  
Male Factor ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Infertile Men ◽  
Significant Difference ◽  
Fisher’S Exact Test

Abstract Background Male factor is the major contributor in roughly half of infertility cases. Genetic factors account for 10–15% of male infertility. Microdeletions of azoospermia factors (AZF) on the Yq region are the second most frequent spermatogenesis disorder among infertile men after Klinefelter syndrome. We detected in our previous study a frequency of 37.5% AZF microdeletions which investigated mainly the AZFb and AZFc. We attempted in this study for the first time to evaluate the frequencies of all AZF sub-regions microdeletions and to analyze reproductive hormonal profiles in idiopathic cases of azoospermic and oligozoospermic men from Sudan. Methods A group of 51 medically fit infertile men were subjected to semen analysis. Four couples have participated in this study as a control group. Semen analysis was performed according to WHO criteria by professionals at Elsir Abu-Elhassan Fertility Centre where samples have been collected. We detected 12 STSs markers of Y chromosome AZF microdeletions using a multiplex polymerase chain reaction. Analysis of reproductive hormone levels including Follicle Stimulating, Luteinizing, and Prolactin hormones was performed using ELISA. Comparisons between outcome groups were performed using Student’s t-test Chi-square test or Fisher’s exact test. Results AZF microdeletion was identified in 16 out of 25 Azoospermic and 14 out of 26 of the Oligozoospermic. Microdeletion in the AZFa region was the most frequent among the 30 patients (N = 11) followed by AZFc, AZFd (N = 4 for each) and AZFb (N = 3). Among the Oligozoospermic participants, the most frequent deletions detected were in the AZFa region (N = 10 out of 14) and was significantly associated with Oligozoospermic phenotype, Fisher's Exact Test (2-sided) p = 0.009. Among the Azoospermic patients, the deletion of the AZFc region was the most frequent (N = 9 out of 16) and was significantly associated with Azoospermia phenotype Fisher's Exact Test p = 0.026. There was a significant difference in Y chromosome microdeletion frequency between the two groups. The hormonal analysis showed that the mean levels of PRL, LH, and FSH in Azoospermic patients were slightly higher than those in oligozoospermic. A weak negative correlation between prolactin higher level and Azoospermic patients was detected. (AZFa r = 0.665 and 0.602, p = 0.000 and 0.0004, AZFb r = 0.636 and 0.409, p = 0.000 and 0.025, and AZFd r = 0.398 and 0.442, p = 0.029 and 0.015). The correlation was positive for AZFa and negative for AZFb and AZFd. Conclusions We concluded in this study that the incidences of microdeletions of the Y chromosome confined to AZF a, b, c and d regions is 58.8% in infertile subjects with 31.4% were Azoospermic and 27.5% were Oligozoospermic. This might provide a piece of evidence that these specified regions of the Y chromosome are essential for controlling spermatogenesis. These findings will be useful for genetic counseling within infertility clinics in Sudan and to adopt appropriate methods for assisted reproduction.

Επίδραση αναλόγου της προστακυκλίνης στην επούλωση αναστομώσεων του παχέος εντέρου επίμυων σε συνθήκες αποφρακτικού ειλεού

2012 ◽  
Author(s):  
Γεώργιος Γαλανόπουλος
Keyword(s):  
Analysis Of Variance ◽  
Anova Analysis ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Significant Difference ◽  
Fisher’S Exact Test

Στόχος της παρούσας διδακτορικής διατριβής ήταν η πειραματική μελέτη της επίδρασης της ιλοπρόστης (ανάλογο της προστακυκλίνης) στην επούλωση αναστομώσεων του παχέος εντέρου επίμυων σε συνθήκες αποφρακτικού ειλεού. Για τη μελέτη χρησιμοποιήθηκαν 80 άρρενες επίμυες, οι οποίοι χωρίστηκαν τυχαιοποιημένα σε 4 (1, 2, 3, 4) ομάδες, με 2 (α, β) ισοδύναμες υποομάδες έκαστη. Στην ομάδα 1 (ελέγχου) και 3 (ιλοπρόστη) διενεργήθηκε τμηματική εντερεκτομή και τελικοτελική αναστόμωση. Στην ομάδα 2 (ειλεός) και 4 (ειλεός και ιλοπρόστη) επιτεύχθηκαν αρχικά συνθήκες αποφρακτικού ειλεού και 24 ώρες μετά διενεργήθηκε τμηματική εντερεκτομή και τελικοτελική αναστόμωση. Η ιλοπρόστη χορηγήθηκε στις ομάδες 3 και 4 σε δόση 2μg/kg Β.Σ. σε 3ml διαλύματος NaCl 0,9% ενδοπεριτοναϊκά, διεγχειρητικά και κάθε ημέρα μέχρι τη θυσία, ενώ αντίστοιχα στις ομάδες 1 και 2 στα πειραματόζωα χορηγούνταν 3ml διαλύματος NaCl 0,9%. Σε κάθε ομάδα τα μισά πειραματόζωα (υποομάδα 1α, 2α, 3α, 4α) θυσιάστηκαν την 4η μετεγχειρητική ημέρα και τα υπόλοιπα (υποομάδα 1β, 2β, 3β, 4β) την 8η. Κατά τη νεκροτομή γινόταν μακροσκοπικός έλεγχος για ρήξη της αναστόμωσης, ύπαρξη περιτονίτιδος ή περιαναστομωτικού αποστήματος καθώς και ποσοτική αξιολόγηση των συμφύσεων σύμφωνα με την κλίμακα Van der Hamm. Ακολουθούσε μέτρηση της πίεσης διάσπασης και στη συνέχεια τμήμα της αναστόμωσης αποστέλλονταν για ιστολογική εξέταση κατά την οποία αξιολογούνταν η φλεγμονώδης αντίδραση (διήθηση από ουδετερόφιλα), η νεοαγγειογένεση, ο αριθμός των ινοβλαστών και η εναπόθεση νεοκολλαγόνου. Η ταξινόμηση των μικροσκοπικών ευρημάτων έγινε σύμφωνα με την κλίμακα Ehrlich και Hunt με τις τροποποιήσεις κατά Phillips. Επιπλέον, προσδιορίστηκε βιοχημικά η συγκέντρωση υδροξυπρολίνης και κολλαγενάσης I επί της αναστόμωσης. Για την συνοπτική παρουσίαση των αποτελεσμάτων υπολογίστηκαν απόλυτες και σχετικές συχνότητες (ποσοστά %), δείκτες κεντρικής τάσης (μέσοι όροι, διάμεσες τιμές) και δείκτες διασποράς (ελάχιστες τιμές, μέγιστες τιμές, τυπικές αποκλίσεις). Για τη σύγκριση των μέσων όρων χρησιμοποιήθηκε το κριτήριο της Ελάχιστης Σημαντικής Διαφοράς (Least Significant Difference-LSD), μετά από την εφαρμογή της μεθόδου ANOVA (Analysis of Variance). Για τις συγκρίσεις των ποσοστών, εφαρμόστηκε ο ακριβής έλεγχος του Fisher (Fisher’s Exact Test). Από την ανάλυση των πειραματικών δεδομένων προέκυψε ότι η ενδοπεριτοναϊκή χορήγηση ιλοπρόστης σε συνθήκες αποφρακτικού ειλεού, έχει ως αποτέλεσμα τον περιορισμό της αρνητικής δράσης του ειλεού στην επούλωση των αναστομώσεων του παχέος εντέρου. Συγκεκριμένα, την 4η και 8η μετεγχειρητική ημέρα ελαττώνει σημαντικά την απώλεια σωματικού βάρους. Επίσης, προάγει τη νεοαγγειογένεση, ενώ συγχρόνως αυξάνει τον πολλαπλασιασμό των ινοβλαστών και τη συγκέντρωση υδροξυπρολίνης. Επιπλέον, την 4η μετεγχειρητική ημέρα ελαττώνει τη φλεγμονώδη αντίδραση και μειώνει τη συγκέντρωση κολλαγενάσης Ι. Σταδιακά, την 8η μετεγχειρητική ημέρα αυξάνει τη σύνθεση νεοκολλαγόνου στην περιοχή της αναστόμωσης. Οι παραπάνω δράσεις έχουν ως αποτέλεσμα την αύξηση της μηχανικής ισχύος των αναστομώσεων, κατά την 4η και 8η μετεγχειρητική ημέρα, όπως αυτή προκύπτει από τη μέτρηση των πιέσεων διάσπασης. Συμπερασματικά, η άμεση μετεγχειρητική ενδοπεριτοναϊκή χορήγηση ιλοπρόστης ενισχύει τους μηχανισμούς επούλωσης και αντισταθμίζει την αρνητική δράση του ειλεού στην επούλωση των αναστομώσεων του παχέος εντέρου.

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