Embryotoxicity testing of IVF disposables: how do manufacturers test?

2020 ◽  
Vol 35 (2) ◽  
pp. 283-292 ◽  
Author(s):  
L Delaroche ◽  
P Oger ◽  
E Genauzeau ◽  
P Meicler ◽  
F Lamazou ◽  
...  
Keyword(s):  
Culture Media ◽  
Toxicity Testing ◽  
Human Sperm ◽  
Culture Conditions ◽  
Mouse Strains ◽  
Control Programs ◽  
Registration Number ◽  
Competing Interest ◽  
High Degree

Abstract STUDY QUESTION How do manufacturers perform embryotoxicity testing in their quality control programs when validating IVF consumables? SUMMARY ANSWER The Mouse Embryo Assay (MEA) and Human Sperm Survival Assay (HSSA) used for IVF disposables differed from one manufacturer to another. WHAT IS KNOWN ALREADY Many components used in IVF laboratories, such as culture media and disposable consumables, may negatively impact human embryonic development. STUDY DESIGN, SIZE, DURATION Through a questionnaire-based survey, the main manufacturers of IVF disposable devices were contacted during the period November to December 2018 to compare the methodology of the MEA and HSSA. We focused on catheters for embryo transfer, catheters for insemination, straws, serological pipettes, culture dishes and puncture needles used in the ART procedures. PARTICIPANTS/MATERIALS, SETTING, METHODS We approached the manufacturers of IVF disposables and asked for details about methodology of the MEA and HSSA performed for toxicity testing of their IVF disposable devices. All specific parameters like mouse strains, number of embryos used, culture conditions (media, temperature, atmosphere), extraction protocol, subcontracting, and thresholds were registered and compared between companies. MAIN RESULTS AND THE ROLE OF CHANCE Twenty-one companies were approached, of which only 11 answered the questionnaire. Significant differences existed in the methodologies and thresholds of the MEA and HSSA used for toxicity testing of IVF disposables. Importantly, some of these parameters could influence the sensitivity of the tests. LIMITATIONS, REASONS FOR CAUTION Although we approached the main IVF manufacturers, the response rate was relatively low. WIDER IMPLICATIONS OF THE FINDINGS Our study confirms the high degree of heterogeneity of the embryotoxicity tests performed by manufacturers when validating their IVF disposable devices. Currently, no regulations exist on this issue. Professionals should call for and request standardization and a future higher degree of transparency as regards embryotoxicity testing from supplying companies; moreover, companies should be urged to provide the users clear and precise information about the results of their tests and how testing was performed. Future recommendations are urgently awaited to improve the sensitivity and reproducibility of embryotoxicity assays over time. STUDY FUNDING/COMPETING INTEREST(S) This study did not receive any funding. L.D. declares a competing interest with Patrick Choay SAS. TRIAL REGISTRATION NUMBER N/A

scholarly journals Uterus transplantation: joys and frustrations of becoming a ‘complete’ woman—a qualitative study regarding self-image in the 5-year period after transplantation

2020 ◽  
Vol 35 (8) ◽  
pp. 1855-1863 ◽  
Author(s):  
Stina Järvholm ◽  
Anders Enskog ◽  
Catrina Hammarling ◽  
Pernilla Dahm-Kähler ◽  
Mats Brännström
Keyword(s):  
Small Sample Size ◽  
Small Sample ◽  
Post Transplantation ◽  
Uterus Transplantation ◽  
Self Image ◽  
Registration Number ◽  
Competing Interest ◽  
The Government

Abstract STUDY QUESTION How is a women’s self-image affected by uterus transplantation (UTx)? SUMMARY ANSWER Women experienced receiving a uterus in both positive and negative ways, but in general, their self-image was positively affected; regardless of whether they have given birth to a child or not, recipients describe themselves as being ‘back to normal’ after the hysterectomy to remove the transplanted uterus. WHAT IS KNOWN ALREADY UTx has repeatedly proved to be a successful treatment for absolute uterine factor infertility. However, there has been no previous qualitative long-term research into the self-image of women undergoing UTx. STUDY DESIGN, SIZE, DURATION This complete, prospective cohort study included the nine recipients of the first UTxs performed in Sweden mostly in 2013. Interviews took place in the 5 years following surgery. PARTICIPANTS/MATERIALS, SETTING, METHODS Eight out of the nine recipients had congenital absence of the uterus, a characteristic of Mayer–Rokitansky–Küster–Hauser syndrome, and one recipient lacked a uterus after a radical hysterectomy due to cervical cancer. The mean age of participants was 31.5 years at inclusion and at this time they all lived in stable marital relationships. Post-transplantation, interviews were performed annually for 5 years, comprising a total of 43 interviews. The interview followed a semi-structured guide. All interviews (median duration of around 25 minutes) were recorded, transcribed verbatim and then analysed by thematic approach. MAIN RESULTS AND THE ROLE OF CHANCE The joys and frustrations of becoming a ‘complete’ woman are seen as a master theme, which influences the three underlying subthemes, a changed self-perception, a changed body and a changed sexuality. Each of these subthemes have three underlying categories. LIMITATIONS, REASONS FOR CAUTION The small sample size is a limitation. WIDER IMPLICATIONS OF THE FINDINGS The results provide information that will be helpful in pre-operative screening procedures and in the psychological support offered both to women who experienced successful and unsuccessful outcomes following UTx. STUDY FUNDING/COMPETING INTEREST(S) Funding was received from the Jane and Dan Olsson Foundation for Science; the Knut and Alice Wallenberg Foundation; an ALF grant from the Swedish state under an agreement between the government and the county councils; the Swedish Research Council; a Ferring Pharmaceuticals scholarship in memory of Robert Edwards; and the Iris Jonzén-Sandblom and Greta Jonzén Foundation. The authors have no competing interests. TRIAL REGISTRATION NUMBER NCT01844362.

scholarly journals Peri-implantation intercourse does not lower fecundability

2020 ◽  
Vol 35 (9) ◽  
pp. 2107-2112
Author(s):  
Joseph B Stanford ◽  
Jared L Hansen ◽  
Sydney K Willis ◽  
Nan Hu ◽  
Alun Thomas
Keyword(s):  
Cycle Length ◽  
Time Window ◽  
Credible Interval ◽  
Cervical Mucus ◽  
Sensitivity Analyses ◽  
Fertility Awareness ◽  
Implantation Time ◽  
Registration Number ◽  
Competing Interest

Abstract STUDY QUESTION Does sexual intercourse in the implantation time window (5–9 days after ovulation) reduce fecundability? SUMMARY ANSWER After adjustment for intercourse in the fecund window and clustering by couple, there was no association between intercourse in the implantation time window and fecundity. WHAT IS KNOWN ALREADY Previous research has suggested an association between intercourse in the peri-implantation time window (5–9 days after estimated ovulation) and reduced fecundability. STUDY DESIGN, SIZE, DURATION We used data from the FERTILI study, a prospective observational study conducted in five European countries, with data collected from 1992 to 1996. PARTICIPANTS/MATERIALS, SETTING, METHODS Women who were experienced in fertility awareness tracking kept a daily diary of cervical mucus observations, basal body temperature measurements, coitus and clinically identified pregnancy. We estimated the day of ovulation as cycle length minus 13 days. From 661 women, 2606 cycles had intercourse during the fecund window (from 5 days before to 3 days after the estimated day of ovulation), resulting in 418 pregnancies (conception cycles). An established Bayesian fecundability model was used to estimate the fecundability ratio (FR) of peri-implantation intercourse on fecundability, while adjusting for each partner’s age, prior pregnancy, the couple’s probability of conception and intercourse pattern(s). We conducted sensitivity analyses estimating ovulation as cycle length minus 12 days, or alternatively, as the peak day of estrogenic cervical mucus. MAIN RESULTS AND THE ROLE OF CHANCE There was no effect of peri-implantation intercourse on fecundability: adjusted FR for three or more acts of peri-implantation intercourse versus none: 1.00, 95% credible interval: 0.76–1.13. Results were essentially the same with sensitivity analyses. There was an inverse relationship between frequency of intercourse in the fecund window and intercourse in the peri-implantation window. LIMITATIONS, REASONS FOR CAUTION Women with known subfertility were excluded from this study. Many couples in the study were avoiding pregnancy during much of the study, so 61% of otherwise eligible cycles in the database were not at meaningful risk of pregnancy and did not contribute to the analysis. Some couples may not have recorded all intercourse. WIDER IMPLICATIONS OF THE FINDINGS We believe the current balance of evidence does not support a recommendation for avoiding intercourse in the peri-implantation period among couples trying to conceive. STUDY FUNDING/COMPETING INTEREST(S) No external funding. The authors have no potential competing interests. TRIAL REGISTRATION NUMBER N/A.

P–264 Clinical relevance of re-expansion after blastocyst thawing

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Aparici. González ◽  
L Herrer. Grassa ◽  
L Cascale. Romero ◽  
J Lláce. Aparicio ◽  
J Te. Morro ◽  
...  
Keyword(s):  
Clinical Outcomes ◽  
Live Birth Rate ◽  
Clinical Results ◽  
Blastocyst Stage ◽  
Good Prognosis ◽  
Culture Conditions ◽  
Miscarriage Rate ◽  
Registration Number ◽  
The Subject

Abstract Study question Are there any differences in clinical outcomes after SET of re-expanded versus non-re-expanded blastocysts? Summary answer The transfer of re-expanded thawed blastocysts is associated with improved clinical outcomes. What is known already Improvements in embryo culture conditions, endometrial receptivity protocols and vitrification as a revolutionary cryopreservation technique have allowed the expansion of blastocyst stage transfers (Lieberman and Tucker, 2006; Stanger et al., 2012; Rienzi et al., 2017), increasing clinical pregnancy and implantation rates in IVF cycles. The re-expansion of thawed blastocyst at the time of transfer has been considered as a good prognosis factor, but not always thawed embryos re-expand. To evaluate the relevance of this event, we compared the clinical results of the re-expanded embryos versus the collapsed ones after their thawing and transfer. Study design, size, duration A total number of 1.125 frozen-thawed blastocyst transfers were included in this retrospective observational study between January 2018 and December 2020. Seven hundred and eighty-six thawed blastocyst were fully expanded at the time of the transfer and 339 thawed blastocysts were non-re-expanded when they were transferred. Participants/materials, setting, methods 1.125 single frozen-thawed blastocyst embryo transfer (SET) cycles (802 from donated and 319 from autologous oocytes) were divided in two groups (re-expanded vs non-re-expanded). Positive beta human chorionic gonadotrophin (bHCG), pregnancy rate (PR), early miscarriage rate (EMR) and live birth rate (LBR) were compared between the two groups. Blastocysts were thawed using an Irvine Scientific® Thaw kit, Irvine Scientific® and were transferring in culture medium (Global® Total® LP, CooperSurgical®). Main results and the role of chance During 2018, 190 re-expanded blastocyst and 94 non-re-expanded were transferred. Statistical significant differences were found in the percentage of positive bHCG (48.4% vs 30.9%, p < 0,0048) and PR (39.5% vs 25.5%, p < 0,0203), respectively. In 2019, statistical differences were found in the LBR between 307 re-expanded blastocyst and 124 non-re-expanded (30.6% vs 12.9%; p < 0,00001). Differences were also found in positive bHCG (50.2% vs 21.8%, p < 0,00001) and PR (40.7% vs 15.3%, p < 0,00001), respectively. Finally, in 2020, 289 re-expanded blastocyst and 121 non-re-expanded were transferred, and significant differences were obtained in the percentage of positive bHCG (46.8% vs 22.3%, p < 0,00001) and PR (32.9% vs 15.7%, p < 0,00001), respectively. Globally, all the variables analysed were statistically significant in favour of the re-expanded embryo group: positive bHCG (48.7% vs 24.5%; p < 0,00001), PR (37.5% vs 18.3%; p < 0,00001) and LBR (20.1% vs 9.5%; p < 0,00001), except for EMR. Limitations, reasons for caution The inherent limitations to a retrospective design. Larger studies are warranted in order to reach robust conclusions on the subject. Wider implications of the findings: Transfer of re-expand blastocyst could be a positive indicator of clinical outcomes. In case of non-re-expand embryos, transfer of two could be reasonable. Trial registration number NONE

Possible Role of Xanthobaccins Produced byStenotrophomonas sp. Strain SB-K88 in Suppression of Sugar Beet Damping-Off Disease

1999 ◽  
Vol 65 (10) ◽  
pp. 4334-4339 ◽  
Author(s):  
Takato Nakayama ◽  
Yoshihisa Homma ◽  
Yasuyuki Hashidoko ◽  
Junya Mizutani ◽  
Satoshi Tahara
Keyword(s):  
Sugar Beet ◽  
Culture Media ◽  
Culture Fluid ◽  
Culture Conditions ◽  
Test Tube ◽  
Disease Suppression ◽  
Antifungal Compounds ◽  
Damping Off ◽  
Plant Basis

ABSTRACT Three antifungal compounds, designated xanthobaccins A, B, and C, were isolated from the culture fluid of Stenotrophomonassp. strain SB-K88, a rhizobacterium of sugar beet that suppresses damping-off disease. Production of xanthobaccin A in culture media was compared with the disease suppression activities of strain SB-K88 and less suppressive strains that were obtained by subculturing. Strain SB-K88 was applied to sugar beet seeds, and production of xanthobaccin A in the rhizosphere of seedlings was confirmed by using a test tube culture system under hydroponic culture conditions; 3 μg of xanthobaccin A was detected in the rhizosphere on a per-plant basis. Direct application of purified xanthobaccin A to seeds suppressed damping-off disease in soil naturally infested by Pythiumspp. We suggest that xanthobaccin A produced by strain SB-K88 plays a key role in suppression of sugar beet damping-off disease.

P-180 Bisphenols are present in culture media used for ART and cell culture

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Vignault ◽  
A Togola ◽  
A Desmarchais ◽  
O Téteau ◽  
V Maillard ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bisphenol A ◽  
Solid Phase ◽  
Culture Media ◽  
Oocyte Quality ◽  
Isotopic Dilution ◽  
Bisphenol S ◽  
Registration Number ◽  
On Line

Abstract Study question Do plastic laboratory consumables and cell culture media used in human ART contain bisphenols? Summary answer Human embryo development media contained bisphenols close to the nanomolar concentration range while no release of bisphenols by plastic consumables was detected under routine conditions. What is known already The deleterious effect of the endocrine disruptor bisphenol A (BPA) on female fertility raised concerns regarding ART outcome. BPA was detected neither in media nor in the majority of plastic consumables used in ART, however it might have already been replaced by its structural analogs, including bisphenol S (BPS). Study design, size, duration Seventeen plastic consumables and 18 cell culture and ART media were assessed for the presence of bisphenols. Participants/materials, setting, methods Ten different bisphenols (bisphenol A, S, AF, AP, B, C, E, F, P, and Z) were measured using an isotopic dilution according to an on-line solid phase extraction / liquid chromatography/mass spectrometry method. Main results and the role of chance While all the plastic consumables of this study did contain bisphenols, none of them did release bisphenols under routine conditions. Moreover, 16 of the 18 cell culture and ART media assessed contained bisphenols, including 8 among the 10 media used in human ART. Five human ART media exhibited bisphenol concentrations higher than 0.8 nM and reached up to 3.2 nM (799 ng/L). Limitations, reasons for caution Further studies are required to investigate a greater number of ART media to identify less potentially harmful ones, in terms of bisphenol content. Wider implications of the findings As BPS has already been reported to impair oocyte quality at nanomolar concentrations, its presence in ART media, at a similar concentration range, could contribute to a decrease in the ART success rate. Thus far, there has been no regulation of these compounds in the ART context. Trial registration number Not applicable

P–180 Bisphenols are present in culture media used for ART and cell culture

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Vignault ◽  
A Togola ◽  
A Desmarchais ◽  
O Téteau ◽  
V Maillard ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bisphenol A ◽  
Solid Phase ◽  
Culture Media ◽  
Oocyte Quality ◽  
Isotopic Dilution ◽  
Bisphenol S ◽  
Registration Number ◽  
On Line

Abstract Study question Do plastic laboratory consumables and cell culture media used in human ART contain bisphenols? Summary answer Human embryo development media contained bisphenols close to the nanomolar concentration range while no release of bisphenols by plastic consumables was detected under routine conditions. What is known already The deleterious effect of the endocrine disruptor bisphenol A (BPA) on female fertility raised concerns regarding ART outcome. BPA was detected neither in media nor in the majority of plastic consumables used in ART, however it might have already been replaced by its structural analogs, including bisphenol S (BPS). Study design, size, duration Seventeen plastic consumables and 18 cell culture and ART media were assessed for the presence of bisphenols. Participants/materials, setting, methods Ten different bisphenols (bisphenol A, S, AF, AP, B, C, E, F, P, and Z) were measured using an isotopic dilution according to an on-line solid phase extraction / liquid chromatography/mass spectrometry method. Main results and the role of chance While all the plastic consumables of this study did contain bisphenols, none of them did release bisphenols under routine conditions. Moreover, 16 of the 18 cell culture and ART media assessed contained bisphenols, including 8 among the 10 media used in human ART. Five human ART media exhibited bisphenol concentrations higher than 0.8 nM and reached up to 3.2 nM (799 ng/L). Limitations, reasons for caution Further studies are required to investigate a greater number of ART media to identify less potentially harmful ones, in terms of bisphenol content. Wider implications of the findings: As BPS has already been reported to impair oocyte quality at nanomolar concentrations, its presence in ART media, at a similar concentration range, could contribute to a decrease in the ART success rate. Thus far, there has been no regulation of these compounds in the ART context. Trial registration number Not applicable

scholarly journals Voltage-gated proton channels exist in the plasma membrane of human oocytes

2019 ◽  
Vol 34 (10) ◽  
pp. 1974-1983 ◽  
Author(s):  
R Ya Smith ◽  
D Morgan ◽  
L Sharma ◽  
V V Cherny ◽  
N Tidswell ◽  
...  
Keyword(s):  
Large Scale ◽  
Human Sperm ◽  
Proton Channel ◽  
Human Oocytes ◽  
Proton Channels ◽  
Voltage Gated ◽  
Registration Number ◽  
Heterologous Expression Systems ◽  
Competing Interest ◽  
Proton Currents

Abstract STUDY QUESTION Do human oocytes express voltage-gated proton channels? SUMMARY ANSWER Human oocytes exhibit voltage-gated proton currents. WHAT IS KNOWN ALREADY Voltage-gated proton currents have been reported in human sperm, where they contribute to capacitation and motility. No such studies of human oocytes exist. STUDY DESIGN, SIZE, DURATION Voltage-clamp studies were undertaken using entire oocytes and vesicles derived from oocytes and in excised patches of membrane from oocytes. PARTICIPANTS/MATERIALS, SETTING, METHODS Frozen, thawed human metaphase II oocytes were obtained from material donated to the gamete repository at the Rush Center for Advanced Reproductive Care. Prior to patch clamping, oocytes were warmed and equilibrated. Formation of an electrically tight seal requires exposing bare oolemma. Sections of the zona pellucida (ZP) were removed using a laser, followed by repeated pipetting, to further separate the oocyte from the ZP. Patch-clamp studies were performed using the whole-cell configuration on oocytes or vesicles derived from oocytes, and using inside-out patches of membrane, under conditions optimized to detect voltage-gated proton currents. MAIN RESULTS AND THE ROLE OF CHANCE Proton currents are present at significant levels in human oocytes where they exhibit properties similar to those reported in other human cells, as well as those in heterologous expression systems transfected with the HVCN1 gene that codes for the voltage-gated proton channel. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION Human oocytes are large cells, which limits our ability to control the intracellular solution. Subtle effects of cryopreservation by vitrification and subsequent warming on properties of HVCN1, the HVCN1 gene product, cannot be ruled out. WIDER IMPLICATIONS OF THE FINDINGS Possible functions for voltage-gated proton channels in human oocytes may now be contemplated. STUDY FUNDING/COMPETING INTEREST(S) NIH R35GM126902 (TED), Bears Care (DM). No competing interests. TRIAL REGISTRATION NUMBER N/A.

scholarly journals Chemosyndrome Variation in Mycobiont Culture and Thallus of Parmelina carporrhizans and Parmelina quercina

2017 ◽  
Author(s):  
David Alors ◽  
Pradeep K. Divakar ◽  
Anjuli Meiser ◽  
Imke Schmitt ◽  
Ana Crespo ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Biosynthetic Pathway ◽  
Culture Media ◽  
Shikimic Acid ◽  
Culture Conditions ◽  
Corn Meal ◽  
Corn Meal Agar ◽  
Artificial Culture ◽  
Acid Pathway

We cultured Parmelina carporrhizans and P. quercina in Corn Meal Agar and 0.2% glucose Malt Yeast Agar for 160 days. Chemosyndrome of natural thalli and mycobiont cultures were analyzed by HPLC. Lecanoric acid, atranorin, chloratranorin and ergosterol were detected in P. carporrhizans thalli, while lecanoric acid, chloratranorin and aliphates were found in P. quercina thalli. The secondary methabolites pattern between thalli and mycobiont culture was completely different in both species. Both species secreted the phenalenone myeloconone C in culture media and was also detected in P. quercina mycobiont aggregates. Interestingly, the phenolic compounds produced by the mycobiont culture of P. carporrhizans are related to those produced by natural thallus by the same biosynthetic pathway, while the chemosyndrome of P. quercina mycobiont implies switch of biosynthetic pathway from acetate-polymalonate pathway to shikimic acid pathway, with pulvinic acid as major compound of mycobiont culture. The role of Myelochonone C, confluentic acid and pulvinic acid produced by mycobiont culture is discussed as possible adaptive vantage in field as photoprotective agent or as byproduct result of stressing artificial culture conditions.

P–526 Incidence of mosaic embryos in day–6 blastocysts, may late blastulation predispose to mosaicism?

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Cetinkaya ◽  
M A Tufekci ◽  
C Cina. Yapan ◽  
S Kahraman
Keyword(s):  
Culture Media ◽  
High Sensitivity ◽  
Culture Conditions ◽  
Preimplantation Embryos ◽  
Extrinsic Factors ◽  
Chi Square ◽  
Preimplantation Genetic Testing ◽  
Mosaic Embryo ◽  
Thermo Fisher Scientific ◽  
Registration Number

Abstract Study question May the mosaicism ratio be influenced by the time of blastulation in preimplantation genetic testing for aneuploidies (PGT-A)? Summary answer The mosaicism ratio is significantly higher in day–6 blastocysts when compared to day–5 when transferable embryos are considered only (euploids and mosaics). What is known already Conventionally, PGT-A has classified preimplantation embryos as either euploid or aneuploid. Yet, a major improvement in PGT-A methodology, with the introduction of high sensitivity diagnostic Next Generation Sequencing (NGS) technology, has allowed the identification of a third embryo category: the mosaic (Coll et al., 2021). Embryonic mosaicism can be defined as the presence of karyotypically distinct cell lines within an embryo and can be detected by NGS at a rate between 20–80%. In the absence of euploid embryos, mosaic embryos, when transferred, have been shown to deliver healthy live births (PGDIS, 2019). Study design, size, duration This retrospective study was based on 9828 trophectoderm biopsies performed in a single ART clinic between January 2017 and October 2020 for PGT-A cycles with more than one blastocyst. PGT-A cycles with only one blastocyst were excluded because in these cycles’ day–5/day–6 biopsy percentage cannot be calculated. A total number of 8398 and 1430 blastocysts were biopsied on day–5 and day–6, respectively for PGT-A by ReproSeq on Ion Torrent S5 (Thermo Fisher Scientific). Participants/materials, setting, methods Three categories were defined in the PGT-A group with >1 blastocyst biopsied to compare the rate of mosaicism: C1:cycles in which blastocysts were only biopsied on day–5 (n = 1872), C2:99–60% of blastocysts were biopsied on day–5 (n = 483) and C3:0–60% of blastocysts biopsied on day–5 (n = 411). The mean female age (C1:36.0±5.2; C2:35.7±4.8; C3:37.1±4.9) and metaphase II oocytes punctured (C1:9.8±6.5; C2:10.4±5.2; C3:8.4±5.4) were similar and statistically non-significant in all groups. T-test and Chi-square tests were used where appropriate. Main results and the role of chance Overall, from the blastocysts biopsied on day–5 and 6, 35.4% and 25.5% were euploid, 13.7% and 14.7% were mosaic, 50.9% and 59.8% were aneuploid, respectively (p < 0.0001, p = 0.32, p < 0.0001), the mosaicism rate being not statistically different. However, when only transferable blastocysts (euploids and mosaics) were considered (aneuploids being discarded), the rate of mosaic embryos was significantly higher in day–6 when compared to day–5 blastocysts (36.6% vs. 28.0%; p < 0.0001). Morphological blastocyst grading was then investigated: from the day–5 and 6 blastocysts biopsied, 51.5% and 30.8% were of top-quality and 48.5% and 69.2% were of good-quality, respectively. Looking deeper into the categories defined, the euploidy rate was found to be higher in C1 (35.1%) when compared to C2 (34.9%) and C3 (27.7%) (p < 0.0001). The mosaic embryo rate was found to be non-significant when all blastocysts (euploids, aneuploids, mosaics) were considered (C1: 13.7%; C2: 14.1%; C3: 14.2%; p = 0.6492). However, when transferable blastocysts were considered (euploids and mosaics only), the mosaic embryo rate was significantly higher in C3 (33.9%) when compared to C1 (28.1%) and C2 (28.8%) (p = 0.02). For morphological blastocyst grading, regardless of blastulation day, mosaicism was higher for good-quality embryos both when all biopsies and only transferrable embryos were considered (p = 0.0018, p < 0.0001, respectively). Limitations, reasons for caution This study focused on blastocyst formation day and morphological blastocyst grading. Extrinsic factors have also been reported to induce mosaicism: ovarian stimulation, culture media, laboratory and culture conditions, technical issues during the biopsy and sample processing (Munné and Wells, 2017; Katz-Jaffe et al., 2018, Fragouli et al., 2010, 2019). Wider implications of the findings: Mitotic errors during cleavage stage causing mosaicism may lead to lower morphological grade and late blastulation as some cells in those embryos are not diploid thus leading to higher mosaicism for blastocysts that reach blastulation on day6 and/or yield only good-quality embryos. Trial registration number Not applicable

scholarly journals Detection the role of physiological factors to produce carotenoid pigment in Staphylococcus aureus

2019 ◽  
Vol 24 (7) ◽  
pp. 6
Author(s):  
Ghada A. Mohammad ◽  
Suhair Taha Daod
Keyword(s):  
Staphylococcus Aureus ◽  
Ph Value ◽  
Negative Impact ◽  
Culture Media ◽  
Culture Conditions ◽  
Pigment Production ◽  
Carotenoid Pigment ◽  
Physiological Factors ◽  
Solid Media

Three Staphylococcus aureus isolates were used in the present study for extraction and quantitation of carotenoid pigment production. Different culture conditions were used to determine optimum pigmentation such as type of culture media, different pH values, temperature, and finally daily bacterial sub-culturing for more than 2 weeks. Nutrient agar was found to be the best medium with the highest production (1.09). Optimum pH value was 6.5 and gave (1.8) carotenoid. Also results showed that pigment was produced at 37o C more than at 42oC. Daily repeated sub culturing had a negative impact on pigment production. Colonies gradualy lost pigmentation and became white in color. Also, sub-culturing was shown to affect the volume of the colonies as they became smaller on solid media.    http://dx.doi.org/10.25130/tjps.24.2019.122

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