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Volume 18, Issue 6 (June 2020) IJRM 2020, 18(6): 425-438 | Back to browse issues page
 10.18502/ijrm.v13i6.7284
Darbandi S, Darbandi M, Agarwal A, Khorram Khorshid H R, Sadeghi M R, C.Esteves S, et al . Comparing four laboratory three-parent techniques to construct human aged non-surrounded nucleolus germinal vesicle oocytes: A case-control study. IJRM 2020; 18 (6) :425-438
Comparing four laboratory three-parent techniques to construct human aged non-surrounded nucleolus germinal vesicle oocytes: A case-control study
Sara Darbandi1 , Mahsa Darbandi 2, Ashok Agarwal3 , Hamid Reza Khorram Khorshid4 , Mohammad Reza Sadeghi5 , Sandro C.Esteves6 , Pallav Sengupta7 , Sulagna Dutta7 , Zohreh Fathi1 , Hojjat Zeraati8 , Mohammad Mehdi Akhondi1
1- Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
2- Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran. , mahsadarbandi@hotmail.com
3- American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, Ohio, 44195, USA.
4- Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran.
5- Department of Embryology and Andrology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
6- Androfert, Andrology and Human Reproduction Clinic, Campinas, Brazil.
7- Department of Physiology, Faculty of Medicine, Mahsa University, Selangor, Malaysia.
8- Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Keywords: Assisted reproductive techniques, In vitro oocyte maturation techniques, Nuclear transfer techniques, Oocytes, Oocyte donation.
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In vitro maturation (IVM) of human oocytes has contributed to several births withstanding variations in patient disorders (such as premature ovarian failure and polycystic ovary syndrome), oocyte types (such as germinal vesicle [GV] and MI), and culture media. However, IVM success rates in terms of live birth remain low as compared to in vivo maturation (1-3). The insufficiency of suitable culture media and deficiencies in the oocytes due to variations in their developmental capacity might explain the meager effectiveness of IVM in assisted reproduction (3, 4). Moreover, both the in vivo and in vitro embryonic developmental success depends on the initial oocyte status and, especially, the quality of the GV oocyte (5).
The GV oocyte pool comprises two major types, namely, the "not-surrounded nucleolus" (NSN) and the "surrounded nucleolus" (SN) (6). The SN oocytes account for nearly 70-80% of GV oocyte pool and are likely the ones that naturally ovulate and endure the complete embryonic development after both in vivo and IVM and fertilization (6). The NSN oocytes account for the remaining 20-30%, which have stopped growing at the 2- or 4-cell stage after fertilization, as observed in IVM and in vitro fertilization studies [5](7-9).
The oocyte quality is a necessary factor to drive embryonic development via renewal, maturation, fertilization, and implantation (5). Oocyte quality includes numerous functional events like cellular organizations, metabolism, protein translation, DNA transcription, and several other factors that need to be considered (5). Nevertheless, women at advanced reproductive age often have a poor ovarian reserve. As a result, the oocyte cohort obtained after ovarian stimulation is usually limited and of poor quality overall, irrespective of the regimen used for ovarian stimulation (10, 11). The development of methods for the maturation of NSN-GV oocytes is advantageous as they might increase the number of developmentally competent oocytes for in vitro fertilization.
Ten methods have been used to increase the ooplasmic maturation, including nuclear transfer, cytoplasmic (ooplasmic) transfer, allotropic rescue, import of normal RNAs to mitochondria, mitochondrial injection, injection of mtDNA, usage of mitochondrial transcription factor A (TFAM), usage of the inner mitochondrial membrane potential (MMP) and free Ca2+ levels, dietary supplementation with cofactors and vitamins, and gonadotropin injections (12). The nuclear transfer may be performed in three oocyte developmental stages containing the GV, MII, and pronuclear (PN) (12). The two types of OT techniques are synchronous ooplasmic transfer (sOT) and asynchronous ooplasmic transfer (aOT) (4). In this project, germinal vesicle transfer (GVT), sOT, aOT using cryopreserved MII oocytes (caOT), and aOT using waste MII oocytes (waOT) have been used to improve the human aged NSN-GV oocytes' quality.
Our group has been focused on finding ways to improve the growth, viability, and overall quality of immature human aged NSN-oocytes. Our aim has been to find remedies to make such oocytes fit for use in ART thereby allowing the formation of a viable and healthy embryo. In this study, we compare for the first time the effectiveness of four three-parent techniques for the maturation of aged GV oocytes.
1. Source of human oocytes
In this case-control study, the oocytes were provided through female partners of infertile couples who underwent ART treatment at the Avicenna infertility clinic affiliated with Avicenna Research Institute, Tehran, Iran, from September 2016 to September 2017. None of the volunteers had any history of chronic diseases (diabetes, cancer, etc.) or any genital inflammation, chromosomal aberrations, and endocrine abnormality.
The following four types of donated oocytes were utilized in this study from women undergoing intracytoplasmic sperm injection (ICSI) after standardized controlled ovarian stimulation: (1) discarded-aged NSN-GV oocytes obtained from participants aged 38 yr and older, (2) discarded young SN-GV oocyte from participants aged 34 yr and younger, (3) cryopreserved good-quality young MII oocytes from participants aged 34 yr and younger, and (4) waste low-quality young MII oocyte from participants aged 34 yr and younger.
2. Controlled ovarian stimulation, oocyte handling
The cumulus cell-oocyte complexes (COCs) were retrieved from women who underwent controlled ovarian hyperstimulation with recombinant human FSH (r-hFSH, Merck Fertility, Germany) after pituitary suppression with gonadotropin-releasing hormone (GnRH, Merck Fertility, Germany) agonist. The r-hFSH dose was 200 IU/day for the first four days and was adjusted on the fifth day accordingly, if needed, based on the volume and number of ovarian follicles and the serum levels of estradiol (E2). When at least three follicles became 18 mm, the COCs were retrieved transvaginally 36 hr after hCG trigger (5,000 or 10,000 IU) (Merck Fertility, Germany). The cumulus-corona cells (CCs) of COCs were removed chemically by a short-term exposure (the 40 sec) of them to medium (G-MOPS™PLUS, Vitrolife, Sweden) with 80 IU/mL hyaluronidase (Sigma, USA) and repetitious pipetting. Subsequently, all oocytes were considered for development with the inverted microscope at 200× or 400× magnification (13, 14).
3. Oocyte cryopreservation procedures
The cryopreservation protocol involved an oocyte vitrification method. Only MII oocytes were vitrified at about 20ºC based on a protocol (15). The oocytes were placed in the vitrification solution (VS) for 90 sec. According to this, oocytes were equilibrated for 1 min in 20 μl medium (G-MOPS™PLUS, Vitrolife, Sweden). Following oocytes were placed in 20 μl equilibration solution (ES) (Vitrolife, Sweden) with 7.5% (v/v) dimethyl sulphoxide (DMSO) (Sigma, USA) and 7.5% (v/v) ethylene glycol (EG) (Sigma, USA) for 2 min, and after that into another 20 μl ES for 2 min. The oocytes were exposed to the 20 μl ES for 5 min and then into three 20 μl drops of VS composed of 0.5 M sucrose (Sigma, USA), 15% (v/v) DMSO, and 15% (v/v) EG and were then loaded in plastic straws (Cryotpo, Kitazato, Japan) and put into cooled protective straws in liquid nitrogen for cryopreservation. Two or three oocytes were vitrified on one straw and the straws were moved into liquid nitrogen tanks and kept until thawing.
To thaw, the straws were taken from liquid nitrogen tanks and placed in 1-ml 1.0 M sucrose (37ºC) for 1 min. After that, oocytes were moved to 1-ml 0.5 M sucrose for 3 min and to 1 ml medium (G-MOPS™PLUS, Vitrolife, Sweden) for 10 min at 20ºC. Finally, oocytes were washed and cultured in the medium (G-MOPS™PLUS, Vitrolife, Sweden) with 10% serum protein substitute (SPS, IVFonline CT, USA) for 1 hr before manipulation. After 1 hr, the oocytes were assessed. The oocytes with normal size, intact zona pellucida clear perivitelline space, and well plasma membrane were defined as "survived."
4. Experimental design
2.4.1.Three-parent techniques experimental groups
The young fresh SN-GV oocytes, cryo-thawed young MII oocytes, and waste young fresh MII oocytes were used as an auxiliary cell for providing ooplasmic substrates for recipient fresh NSN-GV-aged oocytes. Four experimental groups were established according to the category of oocyte providing this substrate to NSN-GV oocytes as follows: Group A ( GVT, n= 10), Group B (sOT, n= 26), Group C (asynchronous ooplasmic transfer using caOT, n= 12), Group D (asynchronous ooplasmic transfer using waOT, n= 12), and Group E (Control, aged NSN-GV oocytes from participants aged 38 and older, n= 12) (Figure 1 showed the study design).
The developmental stage and status of oocyte were confirmed using the inverted microscope (Diaphot, Nikon Corporation, Tokyo, Japan) at 200× magnification. Oocyte manipulation, GVT, sOT, caOT, and waOT were started within 5 hr after oocyte denudation. A total of 72 oocytes have been investigated in this study.
2.4.2. Germinal vesicle transfer (GVT)
In group A, naked NSN-GV aged-oocytes were incubated in medium (G-MOPS™PLUS, Vitrolife, Sweden) with 7.5 µg/ml cytochalasin B (CB, Sigma, USA), 50 µg/ml 3-isobutyl-1-methylxanthine (IBMX, Sigma, USA) for 1 hr at 37ºC and 6% CO2 (16). Precise localization of nuclear material was performed by moving oocytes in the manipulating medium (G-MOPS™PLUS, Vitrolife, Sweden) containing 12% PVA, 50 µg/ml IBMX, and 7.5 g/ml CB. The removal of nucleus material was carried out "directly" with a little surrounding ooplasm with the aid of a tapered manipulating glass pipette (ID: 25 µm, QG: 45º and θA: 35º) (Dain bioassay Co., Iran). The pipette was inserted through a slit made in the overlying zona pellucida using the XYClone™ laser system (Hamilton Thorne Biosciences, Beverly, MA, USA) and lanced the zona pellucida directly covering the GV (16). The pipette was withdrawn and the pressure inside the holding pipette (OD: 100-150 µm, ID: 30-35 and θA: 30º) was increased gently to eject the GV via the opening. The GV was injected into an enucleated young oocyte subzonally (16). Before the removal of GV, to reduce cohesion of the membranes within the pipette, the pipettes were rinsed in polyvinyl pyrrolidone 10% (PVP, Sigma, USA). Following two mineral oil droplets were used to control the fluid flow during the manipulation (17). Subsequently, the reconstructed oocytes (ROs) were rinsed three times and incubated in medium (G-IVF™ -Vitrolife) at 37ºC for at least 30 min until electrofusion (16, 18). Membrane fusion between oocyte and GV was achieved in a microfusion chamber containing a solution of 0.3 M mannitol (Sigma, USA), 0.1 mM calcium chloride (Sigma, USA), 0.05 mg/mL human serum albumin (Sigma, USA), and 0.1 mM magnesium sulfate (Sigma, USA) at 20ºC. After manual alignment, a single direct current (DC) pulse (70 V/cm DC for 20 µs) was used as a fusion pulse by the use of an ElectroCell Fusion instrument (BTX 2001, BTX, USA) (16). The GVT manipulation of aged NSN-GV oocytes is illustrated in Figure 2. Finally, the oocytes were washed and cultured (G-MOPS™PLUS, Vitrolife, Sweden) with 10% serum protein substitute (SPS, IVF online CT, USA) before ICSI.
2.4.3. Ooplasmic transfer (OT)
Figure 3 showed aged NSN-GV oocytes treated with three methods of OT including sOT, caOT, and waOT. As shown in Figure 3, in synchronous transfer (Group B), the ooplasm of the young SN-GV replaced that of the aged NSN-GV, while in asynchronous transfer (Groups C and D), the replacement of ooplasm was done with young MII oocytes (cryopreserved and wasted, respectively) to an aged oocyte at an NSN-GV stage of development (4, 12). In the OT technique, about 5-15% of ooplasm was removed from the recipient and replaced with donor ooplasm at 37ºC (4, 19). Notably, oocytes received ooplasm with volume more than twice had no reprogramming potential (4, 19). This technique was performed by injecting the donor ooplasm into the recipient by a standard ICSI micropipette (4). The OT techniques were performed on the heated plate (37ºC; THN-60/16 and MS 100 Controller from Linkam Scientific Instruments Ltd., London, UK) of an inverted microscope at 200´ or 400´ magnification (Diaphot, Nikon Corporation, Tokyo, Japan) by the Hoffman Modulation Contrast System (Modulation Optics Inc., Greenvale, NY, USA). The microscope had two coarse positioning manipulators (3D Motor-Driven Coarse control Manipulator MM-188, Narishige) and two three-dimensional hydraulic remote-control micromanipulators (Joystick Hydraulic Micromanipulator MO-188, Narishige, Japan). The oocytes were rinsed and cultured in the medium (G-MOPS™PLUS, Vitrolife, Sweden) with serum protein substitute 10% (SPS, IVF online CT, USA) before ICSI.
2.4.4. Semen preparation
The fresh semen specimens were donated by the normozoospermic male spouse of couples registered in infertility clinic at the Avicenna Research Institute, Tehran, Iran. None of the subjects had any history of azoospermia, leukocytospermia, chronic diseases (diabetes, cancer, etc.), endocrine abnormalities, pelvic and genital infections, Y-chromosome microdeletion, and chromosomal aberrations. The assessment of sperm motility and the count was done based on the approvals of the World Health Organization (20). Sperm parameters were normal when total sperm motility (progressive + nonprogressive) ≥ 40%, normal sperm morphology ≥ 4%, vitality ≥ 58%, and sperm concentration was ≥ 15 million/ml (20). Semen was permitted to liquefy at 20oC for 30 min, and the density gradient centrifugation (DGC) method at 300g for 20 min with PureSperm® (Nidacon, Gothenberg, Sweden) was used to separate mature sperm.
2.4.5. Intracytoplasmic sperm injections and embryo culture
All treated oocytes that reached the MII stage were subjected to sperm injection (16). Only one spermatozoon from the sperm droplet was aspirated, tail-first, into the injection pipette. The oocyte was fixed by the negative pressure of the holding pipette in place of the polar body (PB) was at 12 or 6 o'clock position. The injection pipette was hard-pressed at 3 o'clock to the zona pellucida, the oolemma, and the ooplasm below it while a spermatozoon with about 1-2 picolitres of the medium was inserted into the ooplasm.
The injection pipette was draw back calmly and the injected oocyte was leaved from the holding pipette. Then, the oocytes were washed in medium (G-1™, Vitrolife, Sweden), transferred into droplets of medium (G-1™, Vitrolife, Sweden), covered by lightweight paraffin oil (OVOIL™-Culture oil, Vitrolife, Sweden), and incubated (Heraeus, B5060 EK/O2, Van der Heyden, Brussels, Belgium; 37ºC; 5% O2, 5% CO2, and 90% N2). About 18-20 hr after the sperm injection, the oocytes were assessed under the inverted microscope with a 200´ or 400´ magnification. The extrusion of second PB and formation of pronuclei were assessed too. Fertilization was normal when two clearly intact pronuclei were present. After confirmation of two pronuclei fertilization (2PN), zygotes were cultured in medium (G-1™, Vitrolife, Sweden) covered by paraffin oil (OVOIL™-Culture oil, Vitrolife, Sweden), and incubated in an incubator (Heraeus, B5060 EK/O2, Van der Heyden, Brussels, Belgium; 37ºC; 5% O2, 5% CO2, and 90% N2). Cleavage of two pronuclear (2PN) oocytes was evaluated 24 hr later. The assessment of embryo development was carried out at 48 and 72 hr after the sperm injection procedure (4). The embryo quality was counted according to the homology of the blastomer's size and the number of anucleate fragments (4, 11).
2.4.6. Outcome measures
The primary outcome measures were the rates of fusion (number of reconstructed NSN-GV aged oocytes after manipulation), MI (number of NSN-GV aged oocytes that reached MI stage), MII (number of NSN-GV aged oocytes that reached MII stage), ICSI (number of intact oocytes after ICSI), PN observation (number of pronucleus formation following ICSI), cleavage (number of oocytes that cleaved following ICSI), and percentage of embryos reaching 2-, 4-, and 8-cell stages.
2.5. Ethical consideration
Informed consent was achieved from all volunteers and the use of human oocytes for this study was approved by the Medical Ethics Committee of Avicenna Research Institute (93/4390-93/01/01).
The statistical package for social sciences (SPSS) software (version 22, Chicago, IL, USA) was used for all analyses. Wilcoxon was used for pairwise comparison between groups. The Kaplan-Meier and Log rank were used for evaluation of survival function in nine developmental stages including fusion, MI, MII, ICSI, PN, cleavage, 2-cell, 4-cell, and 8-cell. The "cumulative proportion surviving at end of interval", "probability density", "hazard rate", "number entering interval", "proportion terminating", and "proportion surviving" rate at all developmental stages were compared between groups. The "cumulative proportion surviving at the end of interval" related to the proportion of the cell surviving until the end of the developmental stage. The "number entering interval" showed the denominator cell number for each time interval. The "proportion terminating" represented the proportion of NSN-GV oocytes that had died. The "proportion surviving" represented the proportion of NSN-GV oocytes that had survived during the development. The "probability density" and "hazard rates" for the life table and the height of the survival curve were presented too. In all statistical analyses, a P-value ≤ 0.05 was set as significant. In all tables, four groups of B, C, D, and E were compared and group A has been removed due to lack of changes and only was descripted in the text. The sample size was based on previous studies (21). However, since aged women usually have a low ovarian reserve and most of their retrieved oocytes are used for their own treatment, we found it very hard to obtain these samples for this investigation.
The descriptive and comparative statistics of different methods utilized for improving the fertilization and developmental potential of human aged NSN-GV oocytes are shown in Table I and the life table of different methods utilized for improving the fertilization and developmental potential of human aged NSN-GV oocytes are shown in Table II. In group A, none of the oocytes fused, so this group has been removed from the analysis shown in the tables. In group B, all oocytes fused and 20 achieved the MI stage. Of these, 14 progressed to MII and 8 were fertilized. The pronuclear formation was observed in six resulting zygotes, all of which cleaved and five, four, and three achieved a 2-cell, 4-cell and 8-cell stage, respectively (Tables I, II). In group C, all oocytes fused and achieved the MI stage. Of these, eight progressed to MII and showed signs of fertilization. Of the six cleaved embryos, six, five, and five achieved a 2-cell, 4-cell, and 8-cell stage, respectively (Tables I, II). In group D, all oocytes fused, five and three progressed to MI and MII, respectively, but only one fertilized. This zygote cleaved and reached a 4-cell stage embryo (Tables I, II). In group E, six and two oocytes progressed to MI and MII stages, respectively. Of these, only one fertilized but arrested at the zygote stage (Tables I, II).
The cumulative survival rate, probability density, hazard ratio, the number entering an interval, proportion terminating, and the surviving rate at all developmental stages were compared between groups (Table II). A human aged NSN-GV oocyte that achieved the 8-cell stage development has been illustrated in Figure 4. We found that group C had a higher survival rate than groups B (p < 0.05), D (p < 0.005), and E (p < 0.001) (Table I). Cumulative survival rate at the 8-cell stage was respectively higher in group C (0.42) and B (0.12) than the other groups (Table II). In other words, this gives 42% and 12% probability of surviving up to the 8-cell stage following caOT and sOT, respectively, in groups C and B. The Proportion Surviving represents the proportion of NSN-GV oocytes that survived during the development. Therefore, for groups C and B, the probability of surviving during 8-cell development was 1.00 (approximately 100%) (Table II). The developmental potential of human aged NSN-GV oocytes in groups B, C, D, and E are shown in Figure 5.
To the best of our knowledge, this is the first study to compare the effectiveness of methods aiming at improving the reproductive potential of human aged NSN-GV oocytes. We evaluated four three-parent techniques and found that the caOT technique seems to be advantageous over sOT, waOT, and GVT concerning the developmental capacity of human-aged NSN-GV oocytes. Notwithstanding these promising results, our study has several limitations as the number of NSN-GV oocytes utilized was small due to the inherent difficulties to obtain human oocytes for research.
GVT has been the most studied method to improve oocyte quality. It involves the removal of nuclear chromosomal content from aged oocytes and transplantation into enucleated healthy oocytes (12, 16, 22). In this study, we were not successful in achieving embryo development in any aged NSN-GV RO, owing to the problems in gathering the nuclear structure even after using medium supplemented with IBMX. Studies in other mammal species reported variable success with GVT (16). The discrepancy between our results and those of others might be attributable to the use of different oocyte source. Previous studies lack information concerning the type of GV oocyte utilized - SN or NSN. We speculate that GVT was too invasive for use with NSN-GV oocytes. It has been shown that from mitochondrial dysfunction of aging, oocytes might cause peroxidative damage to nuclear DNA or cell membranes, ultimately resulting in cellular function decay (4, 12). Furthermore, studies reported that some oocytes that had matured in vitro underwent apoptosis (4, 12). It seems that these mechanisms can explain the failure of GVT in aged NSN-GV oocyte.
By contrast, our results suggest that the three other methods might be potentially useful to improve oocyte quality of aged NSH-GV oocytes. As compared with group A (GVT), the rates of oocyte fusion were 100% in groups B (sOT), C (caOT), and D (waOT).
After oocyte fusion, GV oocytes have to progress to the next developmental stages. The extrusion of the first and second polar bodies is used as markers of nuclear maturation and meiosis completion (23, 24). In this study, the mean percentage of oocytes achieving the MI stage was 0% in GVT compared with 76.9% in sOT, 100.0% in caOT, 41.7% in wOT, and 50.0% in the control group (Group E). The ratio of oocytes reaching the MI stage was significantly higher in the sOT and caOT than in the waOT and control oocytes. Likewise, the percentage of oocytes arrived at MII stage was significantly higher in sOT, caOT, and wOT than in the GVT and control groups.
The extrusion of the second PB and formation of pronuclei occur approximately 18-20 hr and 20-24 hr after sperm injections, respectively (12). Our results indicate that the likelihoods of NSN aged-GV-treated oocytes to be fertilized are very low (< 10%) with GVT or wOT methods. By contrast, fertilization rates were highest with caOT (~66%) followed by sOT (~30%), and both methods had higher 2PN fertilization rates than controls (~8%). Notably, in this study, we showed that the 2PN fertilization rate observed with the caOT method was similar than that of achieved after sperm injection of mature oocytes in ART programs. In our study, cleavage zygotes were observed with all but GVT methods. However, asynchronous ooplasmic transfer using cryopreserved MII oocytes [caOT] resulted in more zygotes achieving the 2-cell, 4-cell, and 8-cell stages than the other methods.
Our results indicate that cytoplasmic transfer into human aged NSN-GV oocytes might improve preimplantation development. Considering all maturation and development stages, caOT was the most efficacious method to improve the fertilization and developmental capacity of aged NSN-GV. This approach might increase the reproductive success of aged women with infertility that usually has a limited and poor-quality oocyte cohort available for fertilization.
Although some studies assessed the effects of NT or OT techniques on oocyte maturation (4, 12, 16), to our knowledge, no study has evaluated the effects of these techniques on the quality of aged NSN-GV oocytes. Studies showed that NT or OT are useful to improve ooplasmic maturation in mammalian oocytes, but our result showed that the caOT method was beneficial over sOT, waOT, and GVT in supplementing the developmental capacity of human aged NSN-GV oocytes. It is reported that children born using the OT may exhibit mitochondrial heteroplasmy (12, 25, 26). However, the levels of heteroplasmy were not assessed in this study and this important aspect should be investigated in the 8-cell stage in future studies. The recipient oocyte also has other molecules and organelles that may induce genomics, transcriptomics, proteomics, metabolomics, and epigenomics alterations (12, 27). Along the same lines, the assessment of blastocyst development and embryo genetic and epigenetic status of resulting embryos were not carried out and should be investigated further.
The techniques for improving oocyte quality of mammalian oocytes are evolving. However, there are several gaps in the knowledge that need to be investigated in future research. Besides embryo genetic competence, it is important to investigate the well-being of the offspring generated. There is a risk that genetic and epigenetics modifications are incurred during the application of these methods. RNAs, proteins, mitochondria, associated structures, and energetics substrates are transferred from donor to recipient oocytes (12, 28), and children born using these parent techniques were shown to exhibited mitochondrial heteroplasmy (12). The complex interactions between nuclear DNA and mtDNA as well as with other molecules might cause epigenetic alterations of the nuclear genome which requires close monitoring (28).
The asynchronous ooplasmic transfer using cryopreserved MII oocytes technique (caOT) seems to be advantageous over sOT, waOT, and GVT in the developmental capacity of human-aged NSN-GV oocytes. caOT holds promise concerning its possible role in augmenting the developmental capacity of human aged NSN-GV in terms of fertilization and cleavage, thus warranting further investigation.
This project was financially supported by the Avicenna Research Institute (Tehran, Iran). The authors would like to thank all participants and their family members who voluntarily participated in this study.
All authors declare no financial or commercial competing interest.
Type of Study: Original Article | Subject: Reproductive Biology
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23. Patrizio P, Fragouli E, Bianchi V, Borini A, Wells D. Molecular methods for selection of the ideal oocyte. Reprod Biomed Online 2007; 15: 346-353. [DOI:10.1016/S1472-6483(10)60349-5]
24. Rienzi L, Ubaldi F, Martinez F, Iacobelli M, Minasi MG, Ferrero S, et al. Relationship between meiotic spindle location with regard to the polar body position and oocyte developmental potential after ICSI. Hum Reprod 2003; 18: 1289-1293. [DOI:10.1093/humrep/deg274] [PMID]
25. Chiaratti MR, Meirelles FV, Wells D, Poulton J. Therapeutic treatments of mtDNA diseases at the earliest stages of human development. Mitochondrion 2011; 11: 820-828. [DOI:10.1016/j.mito.2010.11.004] [PMID]
26. Ferreira CR, Burgstaller JP, Perecin F, Garcia JM, Chiaratti MR, Meo SC, et al. Pronounced segregation of donor mitochondria introduced by bovine ooplasmic transfer to the female germ-line. Biol Reprod 2010; 82: 563-571. [DOI:10.1095/biolreprod.109.080564] [PMID]
27. Wolf DP, Mitalipov N, Mitalipov S. Mitochondrial replacement therapy in reproductive medicine. Trends Mol Med 2015; 21: 68-76. [DOI:10.1016/j.molmed.2014.12.001] [PMID] [PMCID]
28. Darbandi M, Darbandi S. Three-Parent IVF Methods in Aged Oocytes and Oocytes with Mitochondrial Problems. Cresco J Reprod Sci 2016; 1: 1-2.


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__ We know that Advent is the season for giving and for loving our neighbor, yet we see our neighbor suffering at this supposed happy time of year. Please click below for more.
_ We are currently in the Advent season as Christians, a time of waiting, prayer and hope. We observe the days leading up to the celebration of Christ's birth and we also take this time to remember that Christ will come again.
We are also in the Holiday season as Americans. No matter how well intentioned, it is nearly impossible to avoid Santa Claus, shopping malls, Amazon.com, and the color red. To celebrate the commercialized Christmas may be wrong, but it is natural for us to feel a sense of excitement and joy as a result of it. Our challenge is to remember to put Christ first, this, and every time of year but also to enjoy the company of family and friends that the season brings.
Yet the month of December is not the brightest time for everyone. The holidays bring memories of loved ones passed, opportunities missed, and connections broken. Depression and high suicide rates increase as the days before Christmas, and then the New Year, draw closer. The supposed celebration and hope we see in and outside the church makes it difficult for many to get through their day-to-day lives this time of year.
Two years ago, while on internship, my stepfather set up a "Blue Christmas Service" for his congregation. The concept was to take time for those who do feel pain during the holiday season so that they may know they do not go through it alone. That no matter what they are suffering from, they have God and God's children to lean on. That they can take the time to grieve and lament about how this time of year can hurt them so much. It is so important for many who are broken that they know they have a strong shoulder to cry on.
Although we at O.S.A. do not have such a service, I would encourage each of you to care for those who are struggling right now. To remember that as we give and receive our gifts, so also, should we remember those who do not have someone to exchange even the gift of love. Jesus was born into a world of hurt and pain and that world continues to go on no matter what month of the year it is. What better way to celebrate our Saviour's birth than to care for another of God's children in their time of need especially if that time is Christmas?


— or whatever you may say in your country !
I am a nature-lover, passionate hobby-baker, experimenting urban gardener, devoted mummy, debutante sewer and possibly a new hopeful for the Olympic Games ( — nope, just kidding on that one ).
So get your backpacks ready and join me for some new adventures!


Let's pretend you're in this situation, One day you wake up a little too late. You have twenty minutes to prepare everything before going to meet a potential client.
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Jack W. Clevinger
Obituary of Jack W. Clevinger
Jack was born in Portland, Oregon to Joseph and Olive Tipton Clevinger. He had two brothers, Richard (Dick) and Don. Jack attended Ainsworth Grade School, Benson High School and then joined the Navy, serving as a Coxswain on the U.S.S. Craven, a destroyer, in the South Pacific during World War II. After the war he worked as a mechanic at Braley and Graham Buick. He met the love of his live, Aileen J. Hedman, in 1951and they married on May 23, 1952. The couple moved to Niagara Falls, NY to stay with Jack's mother. While there, he worked for Bell Laboratories in aeronautical manufacturing. They returned to Portland with their first chid, Tom, after a year. Jack opened a service station at 39th and S.E. Belmont. (It was later purchased by ARCO and is still open in that location.) Three daughters followed, Jill, Judy and Suzi. Jack later went to work for ARCO as a regional manager, followed by positions in sales with two industrial supply companies, Ken R. Humke and A.W. Davis in Portland. They bought a home in Eastmoreland and were members of the Multnomah Athletic Club, active in badminton and tennis. Jack was also a member of the MAC Polar Bears who took a dip in the Polar Bear pool at the Oregon Zoo, or local rivers, each year on New Year's Day. Jack and Aileen bought land in 1968 on the Oregon Coast at Netarts Bay and built the Bay Shore RV Park and Marina. Jack worked full time in Portland and spent his vacations and every weekend, except Christmas, working at their coastal business for 25 years. There was a ten year period where he did not have a vacation. He and Aileen finally hired managers to help them run the bustling business at the Coast. Their boat rental business was busy all summer and Jack was a master at crabbing, clamming and fishing, providing tips to their loyal customers and many friends. After Jack and Aileen sold their business in 1994, they travelled in the winters to Lake Havasu, Arizona and later built a house there. He enjoyed trips to Nevada stopping off to gamble on the way south each year. They enjoyed walking and playing cards with friends and family. Jack loved to dance with Aileen and was always a vibrant presence at every party. They later bought a house in the Arranmore neighborhood near Raleigh Hills in Portland. He and Aileen were members of St. Luke Lutheran Church and for years were active on the Welcoming Committee and with the Grauer Back To School Project to supply low-income children with clothing and shoes at back-to-school time. After Aileen passed away in 2016, Jack bought a condominium in Palm Desert, California. He was happiest when he was in the desert enjoying the sun, watching the golfers go by, and entertaining family and friends. Jack is survived by his brother Don Clevinger, his four children, Tom Clevinger (Becky), Jill Whittaker, Judy Guyer (Skip), Suzi Merz (Kevin) and by his grandchildren Ryan Clevinger, Katie Marsolan (Tim), Greg Sargent, Chloe Hollatz (Tom), Mike Merz, Chris Merz, Dezaray Mullins (Spencer), Madeline Whittaker, William Wilkerson, Jack Merz and J.T. Whittaker, as well as by his great-grandchildren Parker, Paisley and Gibson Hollatz and Daniel and Cynthia Marsolan. Remembrances may be made to the Grauer Back To School Project at St. Luke Lutheran Church at 4595 SW California St., Portland, OR 97219. Service to be held 11:00 AM, Saturday, January 18, 2020, at St Luke Lutheran Church 4595 SW California St. Portland, OR 97219.
St Luke Lutheran
4595 SW California Street
Willamette National Cemetery
11801 SE Mt Scott Blvd.
Cemetery Service

redpajama_set_name: RedPajamaCommonCrawl


Interestingly enough, over the 6 years I've been immersed in the animation world, I've never experienced an animation event. I've been at Sheridan for 4 years now and am officially done school (which is crazy! I'll speak more to that later), and in all that time I've never felt the need to experience TAAFI or any other animation event.
I was given the opportunity to go to TAAFI this past weekend with my friend, Becca (check out her awesome work here!), and it made me realize what a foolish decision it was to pass on it for so long. I only went for one day, but in that day I was able to connect with so many industry professionals in a friendly environment.
When I arrived I was a tad overwhelmed. I'm not at my most confident when it comes to selling myself, as many artists can relate to. We're a breed that prefers to be behind the scenes, more often than not. After settling down, I managed to work up the nerve to chat with some of the people manning the different studio booths, and was met with nothing but positivity.
I was also able to attend two lectures that day. One by Allison Rutland of Pixar (and also a Sheridan Alumni), and one by a panel of short film makers, consisting of Marv Newland, Michael Rianda, Michel Gagne, Rob Silvestri, and Eric Goldberg. As a character animation fanatic, I was obviously most thrilled to hear Eric talk. For those who aren't aware, I was fortunate enough to be mentored by Eric during my time at Disney, and this was the first time I had seen him since my internship. We discussed his role on Moana, as well as his most recent work, this Simpsons Couch gag. It was fantastic getting to reconnect with someone so influential to my career as an artist.
I was actually fortunate enough to see Allison's talk twice, as she also gave a lecture at Sheridan a few days prior. I was pleased to see there were some small differences in each talk, so I was able to get every morsel of information she so kindly provided. Her talk was much more practical than the Birth of a Short Film talk, I found, as it was more about workflow. Allison is an incredible CG animator responsible for the bulk of Riley's emotional scenes in Inside Out, and she was recognized for that by ASIFA when she won an Annie Award this year for her contributions. She has a nuanced understanding of subtlety in animation. I learned a lot about texture, muscle tension, and performance during her talk.
I think the coolest part of TAAFI was in its reminder that the industry is so incredibly small. I saw so many familiar faces, people I've known for years, people I've met once or twice, and a host of Sheridan classmates. It was a really fun time!
Stay tuned for an obligatory "I've graduated!!" post coming soon!
Overcast [Sheridan Third Year Film 2015] from Carly Dziob on Vimeo.
Here's my third year group film!
It was super fun, and we were one of the few groups who essentially went through the whole year without any inter-personal issues.
For those interested in what I animated, the most notable shots would be the opening shot of the rabbit, the two kids looking at the clouds as the camera rotates, the squid exploding the train, the robot deflecting lighting, and the main clouds blowing away all the dark clouds (lets try to not get into the intricacies and politics of the cloud world and why certain clouds are sentient and others aren't). I wish I was able to animate more, but I was also the head of animation for the project, and so my time was largely spent assisting others.
I was mostly found animating on this film, but my biggest responsibility outside of that was cloud character design. Below you'll find some of the vis dev and character design I was responsible for.
So once again, it's been a while. This summer has been a very interesting one for me, as I was given the incredible opportunity to work on my first feature film, while still being able to return to my fourth year at school this September.
I was fortunate enough to be animating on Hotel Transylvania 2 at SPI, where I interned during the summer of my first year at school. It seemed fitting that my first internship gave way to my first real job. Obviously I'm not able to share any of the work I contributed quite yet, but be sure to see it in theaters this September! I'll be sure to post my (small) contributions to the film as soon as I'm able.
As far as my third year film, my group and I are involved in several smaller festivals and are waiting on the OK to put the film online. Stay tuned!


Limpopo is the northernmost province of South Africa, sharing its borders with Zimbabwe, Botswana and Mozambique. As such, it has a hotter climate and is home to a different variety of plants and animals; those that can survive these conditions. These are simply spectacular in their beauty and variety, and are a delight to explore and discover.
Staying at one (or more) of Limpopo's game lodges, or in the local bush camps, gives visitors the opportunity to enjoy a comfortable base from which to embark on game drives at dawn and dusk, or even those conducted at night, when nocturnal predators come to life. When staying at a Game Lodge in Limpopo, guests can look forward to seeing lion, cheetah, elephant, rhino, buffalo, giraffe, zebra, ostrich, hyena, and plenty more. The birds are equally impressive, making Limpopo a bird-watcher's heaven.
Limpopo game lodges and camps range from comfortable options that are easy on the bank balance to luxurious retreats that will leave guests feeling pampered and relaxed.


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Komisariat Straży Celnej "Lipnica Wielka" – jednostka organizacyjna Straży Celnej pełniąca służbę ochronną na granicy polsko-czechosłowackiej w latach 1921–1928.

Formowanie i zmiany organizacyjne

Na wniosek Ministerstwa Skarbu, uchwałą z 10 marca 1920 roku, powołano do życia Straż Celną. Od połowy 1921 roku jednostki Straży Celnej rozpoczęły przejmowanie odcinków granicy od pododdziałów Batalionów Celnych. Proces tworzenia Straży Celnej trwał do końca 1922 roku. Komisariat Straży Celnej "Lipnica Wielka", wraz ze swoimi placówkami granicznymi, wszedł w podporządkowanie Inspektoratu Straży Celnej "Żywiec".
W drugiej połowie 1927 roku przystąpiono do gruntownej reorganizacji Straży Celnej. W praktyce skutkowało to rozwiązaniem tej formacji granicznej. Rozporządzeniem Prezydenta Rzeczypospolitej Polskiej Ignacego Mościckiego z 22 marca 1928 roku w jej miejsce powoływano z dniem 2 kwietnia 1928 roku Straż Graniczną.

Służba graniczna
Sąsiednie komisariaty
 komisariat Straży Celnej "Korbielów"  ⇔ komisariat Straży Celnej "Chyżne" − 1926

Funkcjonariusze komisariatu
Obsada personalna w 1926:
 kierownik komisariatu – komisarz Wacław Romiszewski
 pomocnik kierownika komisariatu – starszy przodownik Antoni Krempa (132)

Struktura organizacyjna
Organizacja komisariatu w 1926 roku:
 komenda – Lipnica Wielka
 placówka Straży Celnej "Lipnica Dolna"
 placówka Straży Celnej "Przywarówka"
 placówka Straży Celnej "Krzywoń"
 placówka Straży Celnej "Lipnica Wielka"
 placówka Straży Celnej "Babia Góra"
 placówka Straży Celnej "Zawoja"

Przypisy

Bibliografia

Komisariaty Straży Celnej II Rzeczypospolitej
Formacje graniczne w Lipnicy Wielkiej
Komisariaty Inspektoratu Straży Celnej "Żywiec"

redpajama_set_name: RedPajamaWikipedia


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Confirmation celebrates the full outpouring of the Holy Spirit on a baptised person, completing and strengthening the grace given by the Spirit in Baptism. It marks the Christian as a follower of Christ and a full member of the Church.
In our parish, children normally celebrate Confirmation in Sixth Class. Much of the preparation is carried out in school with the parish offering a complimentary preparation programme.
At Confirmation the Bishop/priest prays that the gifts given by the Holy Spirit for the living out of the Christian life will grow in each person being confirmed. These are the gifts of Wisdom, Understanding, Knowledge, Right Judgement, Courage, Reverence and Wonder and Awe in God's presence. Growth in these gifts is a lifetime's experience and leads the Christian to share in the mission of Jesus Christ. Young people preparing for Confirmation are encouraged to be involved in some form of service in their parish.
The decision to be confirmed is not one to be taken lightly, for it involves both faith and commitment.
In St Canice's Parish the Sacrament is administered by the Priests of the Parish with the blessing of the Bishop mainly due to the very large number of children. This allows for smaller and more intimate celebrations, which also emphasises the Christian Community's involvement.
Who Can Receive?
Any baptised Catholic wishing to advance on the path of developing their faith. For young people, this is usually part of the 5th/6th class primary school programme.
For adults who were not confirmed as children, it means taking part in the Rite of the Christian Initiation of Adults (R.C.I.A.)
A sponsor stands behind the candidate for Confirmation at the Confirmation ceremony and places their hand on the shoulder of the candidate as a sign that they will support them in living out their baptismal promises. However, the role of the sponsor is not just for one day. The sponsor undertakes to assist the confirmed person in growing in the fullness of their faith and in their membership of the Catholic Church.
A person qualifies as a sponsor by being a reasonably mature adult, who is at least 16 years old, and has already received the Sacraments of Initiation, (Baptism, Confirmation and Eucharist) themselves.
The Confirmation sponsor may be one of the people who was a sponsor at Baptism (subject to the notes here).
Choosing a sponsor
To be admitted to undertake the office of sponsor, a person must:
Be appointed by the candidate, or by the parents or whoever stands in their place, or failing these, by the parish priest or the minister; to be appointed the person must be suitable for this role have the intention of fulfilling it.
Be not less than sixteen years of age, unless a different age has been stipulated by the diocesan Bishop, or unless the parish priest or the minister considers that there is a just reason for an exception to be made.
Be a Catholic who has been confirmed and has received the blessed Eucharist, and who lives a life of faith which befits the role to be undertaken.
Not labor under a canonical penalty, whether imposed or declared.
Not be either the father or the mother of the candidate.
The tradition of taking a new name at Confirmation emphasises the new identity of a Christian being called to witness to their faith. People are encouraged to take the name of a saint or a person from the Bible who inspires them in some way.
The oil used is called the Oil of Chrism. It is olive oil mixed with balsam that is blessed by the bishop and priests of the diocese in the Cathedral at the 'Chrism Mass' on Holy Thursday. The Sacrament is conferred with the anointing with this 'Oil of Chrism' on the forehead as the Bishop says 'Be sealed with the gifts of the Holy Spirit'.
Eucharist - First Holy Communion
Penance - Sacrament of Reconciliation


IKEA Billy Bookcase Makeover 04/12/2012 by Diane Henkler 71 Comments I have not one, but two IKEA Billy bookcases that need to be transformed so they coordinate with the new color scheme I am doing my studi office in.
I started by calculating the number of IKEA Billy bookcases needed for this project. It came out to 3 of the 15" wide and 2 of the 31.5" wide shelvings and I bought the 1 foot height extensions as well.
8/05/2011 · So, your left with a wall looking like a colander, now for fun bit; get your bookcase in place and with the (invaluable) aid of an electric screwdriver, fix it good a tight to the wall. It was amazingly solid by the time all the brackets were fastened.


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Tag Archives for People
By Alex Nyarko28/09/2020Articles, Blog, Uncategorized
Celebrating Black Geniuses
I have always been fascinated about the history of science. Learning about the personalities who developed the scientific breakthroughs that we are enjoying now is not just an exhilarating pastime but a source of encouragement…
By Kwaku Nyantakyi20/02/20190News
Breathtaking ScienceSlamGh Comes Off With A Bang
On the 16th of February- 2019-two days after St. Valentine's Day, dozens of academics went to show their love for the very revered field of mathematics at the University of Ghana. The School of Engineering…
By Paul Agbo20/02/20190News
Ghana Establishes Innovation and Research Commercialization Center
The government of Ghana through the Ministry of Environment, Science, Technology and Innovation (MESTI) has launched a center for Innovation and Research Commercialization, Hon. Patricia Appiagyei, deputy minister of the sector, has announced. "This center…
UN International Day for Women and Girls in Sciences
At the just ended UN International Day for Women and Girls in Sciences which was marked at the University of Ghana, it was revealed that Ghana has only 7% Women in Engineering Professions. This was…
By GhScientific04/11/20180News
Entrance University College of Health Sciences Holds First Matriculation
Forty-two students have been admitted to pursue Doctor of Pharmacy programme at the Entrance University College of Health Sciences. The students were outdoored at the Entrance University College of Health Sciences' first inauguration ceremony on…


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Taylor Swift cutout that brightens commuters' mornings will stay despite owner's move
Taylor Swift cutout that brightens commuters' mornings as they ride past her will stay despite owner's move to another city
The much-loved life-sized cutout of the singer has been on display for five years
Red Redmond, 32, displayed 'Tram Taylor' from her window in Manchester
When owner Red Redmond, 32, said they were moving, commuters were upset
Red is looking for a Swiftie to rent the flat so they can look after the cutout
By Gina Kalsi For Mailonline
The owner of a beloved life-sized cutout of Taylor Swift which has brightened people's commute to work will stay, despite them moving to another city.
The life-size likeness of the 'Shake It Off' singer has been displayed from an apartment window in Manchester for five years and is loved by Metrolink tram passengers.
When the flat's owner Red Redmond, 32, announced they were moving to Sheffield, commuters feared a blank space would be left in the cutout's place.
However their love story with the figure, affectionately known as 'Tram Taylor', will continue after hundreds of its lovers rallied to persuade Red to keep it in the flat.
The cut-out, which has its own hashtag on Twitter, quickly became a landmark among city centre commuters as it can be viewed from the Cornbrook Metrolink stop
The life-size likeness of the 'Shake It Off' singer has been displayed from an apartment window in Manchester for five years and is loved by Metrolink tram passengers
Fans even tried to lobby Greater Manchester Mayor Andy Burnham on social media to step in and save the much-loved figure from exile.
Red, a comedy promoter at iconic music venue The Leadmill in Sheffield, is hoping to rent the flat out to a Taylor Swift fan who will look after the cutout.
They said: 'I would obviously love to bring her with me but she's obviously found her home in Manchester and she means a lot to so many people.
'I can't sell my flat because of a cladding issue and I need to rent it out so she'll be staying in the window for the next few months.
'My hope is that we can get someone to rent out the flat who will keep her in.
'It sounds silly but I've had hundreds of direct messages basically requesting to rent the property and keep her in it.
'It's crazy how many people have messaged – I think it's added value to my flat to be honest.'
The singer recently wrapped up 2022 on a high when she made music history by selling six million copies of her Midnights album in just two months.
Commuters took to Twitter post how about how much they will miss the Taylor Swift cutout and to thank Red
Red first displayed the cut-out of the American singer-songwriter in their window in 2017 after being given it at a Taylor Swift-inspired club night at the Leadmill.
The cut-out, which has its own hashtag on Twitter, quickly became a landmark among city centre commuters as it can be viewed from the Cornbrook Metrolink stop.
Red has said that Tram Taylor is 'seen by thousands of people each day' is also used as a 'milestone' for those who go past it on a regular basis.
They said: 'Six months after she had been up, I started to see some tweets of people discussing her online.
'Basically every year since I've had her in the window, it just suddenly goes viral and there will be people tweeting or talking about it.
'She just gets seen by thousands of people every day because she's on their commute to work or people see her on their way home.
'She's kind of like a milestone or a marker for if you're ten minutes from home for example.
'I've heard people say that they'll text each other whenever they pass Taylor Swift because then it means their partner knows they are ten minutes from home.'
Despite its cult status, Red said the cut-out is actually sun damaged and doesn't have any legs from the waist down, but its top half is in 'great condition'.
They added: 'She has had a little bit of damage to her over time.
'She got sun damaged after being in the window for too long and she doesn't have any legs anymore.
'She's been propped up against the window for five years now and the top half of her is in great condition really.'
It has also emerged that Taylor Swift's cat – Olivia Benson – is the world's third richest pet worth a staggering $97million.
Blue star Lee Ryan is found guilty of racially aggravated assault of black female BA attendant
Generation Z reveal which hand gestures they find the least cool


Chicken Hunt Chicken Hunt Shoot all flying chickens.
Mission Commando Mission Commando Shoot the enemies with 3 different guns. Take cover and reload. Take down as many terrorists!
Bottle Shooter Bottle Shooter Shoot 'em down cowboy style! Shoot all bottles as fast and many as you can before the time runs out!
007 Agent Attack 007 Agent Attack Shoot the enemies as they pop up. Careful you can scroll left and right.
Description: Play a fan made game of the popular shooting game, Call of Duty. Practice shooting with your high power gun in three scenes from the map Das Arkaden.


text:
Archive for Mississippi
Pro-life march highlights 1st abortion for rape law days before Mother's Day
Posted in Abortion and rape, Abortion History, American Law Institute with tags 50 years, Abortion and rape, abortion history, Ali, American Law Institute, exceptions, First state, Freedom March Life Wins, Juda Myers, liberalize abortion law, Mississippi, Model Penal Code, Pro-Life, rape abortion on April 25, 2016 by saynsumthn
In May, mother's and children of rape conception along with supporters will gather at the Mississippi State Capitol to recognize 50 years of legal abortion in the state which began Mother's Day 1966. The Freedom March Life Wins event, which takes place May 6th is being organized by Juda Myers founder of Choices4Life an organization which supports females who are pregnant after rape and those who are raising the children.
Myers told Live Action Action News that two years ago she heard God tell her to look up the law that legalized abortion in the state of Mississippi. "I looked it up and discovered the law was passed May 8, 1966 [Mother's Day]. So, the Lord told me to go read the text." Myers said she stumbled upon a photo image of the law published by another group who opposes abortion. She discovered that Mississippi was the first state to amended its law to permit abortions in cases of rape. "I felt like someone stabbed me in the heart," Myers said.
Juda Myers
Myers is outspoken against abortion on demand, but she takes a strong stand against rape exceptions because, as she puts it she is one of the rape conceived. At the age of 46, Myers discovered that she was conceived in rape. Two years later she met her birth mother who tearfully recounted how she was gang raped as a young woman and became pregnant with Myers. When the doctors told her mother that they would "take care of it" Myers said that her mother told her she responded by telling the doctors:
" You won't take care of "it" – "it" is MY baby."
Myers discovered that her birth mother lovingly placed her for adoption after feeling pressured to have the abortion. Myers was born on Valentine's Day 1957 and her mom told her when they met that Myers was the best thing that had ever happened in her life. After her mom passed, Myers was told by her close friends that her mother celebrated her birth every Valentine's Day and vowed that they would celebrate together again, which they finally did.
Myers said she is holding the Freedom March Life Wins event a week before Mother's Day, not just for herself but for the mother's who are treated with disrespect like her mother was. Choices4Life reaches out to the rape conceived as well as those who are victims of rape. The stories they tell to Myers has compelled her to be an activist. And Myers, is passionate about her message on abortion for rape: NO EXCEPTIONS!
To Myers and those she works with and represents, abortion for rape is nothing more than genocide against an entire group of people from every race, culture and gender. She points out that when you research the history of abortion in America you will find that model abortion laws originated in the 1950's with the American Law Institute (ALI), an organization of American lawyers whose mission was the reform of American law. They released the first draft of their Model Penal Code to liberalize abortion on May 21, 1959. Although Colorado, California and North Carolina became the first states to adopt the full language of the ALI model penal code, Mississippi was the first state to allow for the rape exception to abortion.
ALI Abortion Model Penal Code as published by the Chicago Tribune May 29. 1966
1966 Attitudes on abortion published by the Chicago Tribune May 29, 1966
Myers points out that people generally refer to children conceived in rape as the "lowest of the low." In fact, not that long ago, a pastor referred to the children of rape as a "demon seed." Myers said that every time she hears about a "rape exception" she gets angered because she knows that those so-called exceptions represent precious preborn children. To Myers, people like her are being targeted for death because of someone else's crime. And, although Myers loves and works within the pro-life movement she says that they are not without criticism for support of such exceptions.
"The pro-life community is willing to use us as a sacrificial negotiating tool, for what they consider to be the greater good."
Myers passionately believes that children conceived in rape are equally as valuable as any other preborn child and should be vigorously protected along with their mothers. Myers believes that because rape was the reason abortion was legalized to begin with, ending rape exceptions can actually end all abortions. She said that Mississippi was the first state to use rape to liberalize their abortion laws but that tactic was also used by Sarah Weddington when she argued the Roe vs. Wade case before the US Supreme Court. The case, which originated in Texas was based on a lie that Norma McCorvey, the Roe in Roe vs. Wade, was raped. McCorvey later recanted her involvement in the case which opened up legal abortion in all 50 states, calling the premise that she was raped a lie.
"It started with rape and it will end with rape. When the rape conceived are valued everyone will have equal value in the room."
Myers said that The Freedom March Life Wins event will honor life conceived in rape. She plans to hold a "Heroes Welcome Reception" on May 5th at the Downtown Marriott in Jackson. There Myers will honor the moms who gave life to their children following a tragic rape. "People who attend are welcome to talk to these moms and hear their stories face to face," she said.
The day following the reception, Myers said they will hold a 9:30 Press Conference inside the state Capital. Then around noon, the rally will kick off with an Honor for Life Award Ceremony where mothers who chose life after rape from around the country will be flown in to be honored. After that, the march, which Myers said will be prayerful, will commence. Myers said that people who are interested in joining her in Jackson, Mississippi can get information about the event at their website.
We march for Freedom! We march for Love! We march for life! Mothers and children of rape conception will show the world that Love Wins when Life Wins!
Get more information here: http://freedommarchlifewins.org/
Pro-life group fights to end stigma of rape conception
Posted in Abortion and rape, Abortion History with tags Abortion, abortion exceptions, church and abortion, First abortion law, Freedom March Life Wins, Juda Myers, Mississippi, pregnant, Pro-Life, rape conceived, rape conception, rape exception, Roe V Wade on November 15, 2015 by saynsumthn
A pro-life group is working to end the stigma of rape conception by organizing a march this spring in one of the first states that legalized abortion. It was 2003 when Juda Myers learned from a social worker that she was conceived in rape. Juda, who was adopted, discovered the brutal truth while looking for her birth mother. The shocking details about the woman who carried her in the womb and how she was gang raped set the stage for Juda's mission to advocate on behalf of the rape conceived.
"The social worker's attitude conveyed centuries of condemnation for the rape conceived," Juda who contemplated suicide after learning the truth told Live Action News. "I was falling into a deep black abyss that brought me to the brink of a breakdown," she said. Juda said her commitment to God kept her from doing the unthinkable, and recounted how a friend who knew nothing of the situation reminded her that God knew her before she was conceived!
In 2005, Juda met her birth mom who was in her 70's at the time, "I walked into the nursing home stood at the desk and was about to ask for Ann when I heard my name called from behind me. I turned to see eyes of crystal blue not unlike my own. I knew it was her. Holding a picture of me when I was only 3 months old at the time of my adoption."
Juda has told her story a number of times pointing out that those who allow abortion exceptions in cases of rape are really saying that she should have never been born. She is passionate about the lives of every preborn child in the womb and said that hearing other moms with similar stories to her mother's experience made her realize that no one was helping moms like them. That revelation inspired Juda to found the non-profit organization Choices4Life.
The mission of helping these mothers and speaking for what many describe as a so-called "small percentages of preborn children" has not been an easy one. Juda has been the target of abusive vitriol from abortion advocates when she posts the stories of young rape victims who choose to carry their babies to term. The so-called advocates of free choice seem to draw a line in the sand if the pregnant female is a young rape victim.
But, the prejudice against the rape conceived extends well beyond abortion advocates and even reaches into the walls of religious institutions. In fact, abortion exceptions for cases of rape were sadly written into main stream denominational resolutions before Justice Harry Blackmun wrote the Roe V. Wade decision, legalizing abortion on demand. In truth, those pro-abortion resolutions helped lay the foundation for the tragic SCOTUS decision.
Since then, church leaders inside some of these denominations have apologized for their positions and recanted the resolutions. But, the bias against the rape conceived remains ongoing. In 2013, a pastor referred to children conceived in rape as a "demon seed." His words shocked the pro-life community and prompted the mother of a child conceived in rape to write a powerful response which read in part:
"For too long, the rape conceived and their mothers have been targets of hate speech. The assumption is that no woman who was REALLY raped would want to carry such a baby to term. That is a lie. Using derogatory language to the woman about her child often coerces her into making a decision that puts her at risk mentally, emotionally, and physically. She may consent to an abortion in order to prove that she was actually raped or out of fear that her child will be the object of continued prejudice after birth."
Choices4Life works tirelessly to educate society that children conceived in rape are not the criminals and have done nothing wrong. "To condemn a human for the crime of another is not justice," Juda often points out. But rape is the cornerstone of abortion and was the lie that brought the case to the Supreme Court in 1973. It is the "necessary evil" that continues to keep abortion legal today. Juda says it is time for the pro-life community to rally around the victims of rape and not give the perception that they are expendable simply because they may be few in numbers.
Juda and her Choices4Life organization contend that if we destroy this leg of abortion deception we may very well end abortion in America. To chip away at the foundation of Roe, Choices4Life is organizing the FREEDOM MARCH #LIFEWINS event next May in Mississippi. According to the event website, the first abortion law was passed in Mississippi on May 8, 1966 and it was for cases of rape and incest. Ironically, this year, May 8th happens to fall on Mother's Day 2016.
In addition to the march, Choices4Life is organizing what they call a "hero's welcome" reception to "these amazing mothers who saved the life of their children conceived in rape." The hope is that listening to the stories of the brave women who rejected the lie that a child they created is worthless simply because of the crime of their fathers, will transform those who attend.
"In our day tolerance is shouted on every street corner. It's time to end the stigma of rape conception and rally around these heroic mothers who fight for the life of their baby conceived in rape. Mothers and children are not guilty of any crime and don't deserve to be treated as criminals," Juda writes in the Freedom March Life Wins press release.
CHOICES4LIFE helps pregnant rape and incest victims with relocation assistance, they have subsidized rent, paid utility and medical bills and offered housing in private homes for mothers in need. The non profit has reached out to victims of all races, religious and sexual orientation ranging in ages from 10 to 42 years of age. Juda has dedicated her work to her birth mother whom she calls "a Hero." Juda lost her Hero a few years ago but is grateful God allowed them to find each other. She tells those who support Choices4Life and women she reaches out to, "Be a hero.
"A hero is defined as saving a life," she says.
Get more information about Choices4Life here.
Abolitionists in Mississippi protest abortion at churches / festival
Posted in AHA with tags Abolish Human Abortion, Abortion, AHA, Anti-abortion, Christian Concert, Church, Jackson, Mississippi, Ninevah on April 2, 2015 by saynsumthn
Abolish Human Abortion is in Jackson protesting more than the last remaining abortion clinic in Mississippi.
Watch and feel free to comment:
Drop Carding the mall:
Convention Center for Christian concert:
Festival:
Black abortion doc receives award named after Klan speaker Margaret Sanger
Posted in Abortionist, Abortionist Racist, Black abortionist, Margaret Sanger Award, Planned Parenthood Maggie Awards, Planned Parenthood Margaret Sanger Award, Planned Parenthood uses blacks, Willie Parker with tags Abortion, Abortionist, Black abortionist, Dr. Willie Parker, Eugenics, Margaret Sanger Award, Mississippi, Planned Parenthood, Planned Parenthood Metropolitan Washington, racist, Willie Parker on March 30, 2015 by saynsumthn
Dr. Willie Parker is no stranger to controversy. He is a circuit riding abortionist who, most notably works at the last remaining abortion clinic in Mississippi.
Parker has made many outrageous statements claiming that he performs abortions because he is a "Christian" despite the long-held view that murdering a baby in the womb is the antithesis of Christ-like.
Parker recently claimed that NOT PROVIDING ABORTIONS was RACIST.
Yes, you heard me correctly.
As a result of Parker's unwavering stand for abortion, and his dedication to killing majority Black babies, Planned Parenthood recently "honored" the abortionist during Black History Month 2015 even placing abortionist Willie Parker on their list of Dream Keeper "Doers."
But, the organization based on eugenics and racism did not stop there.
At their 2015 Gala, Planned Parenthood, which took place March 19th, Planned Parenthood gave the black abortionist their most prestigious award, The Margaret Sanger award.
Sanger was the founder of Planned Parenthood who admitted giving Klan speeches in her autobiography.
The Klan would be right in line with Sanger's views on limiting the births of the so-called feeble minded and unfit. The eugenics groups she belonged to and placed on her board as well as published in her Birth Control Review, also held those same racist views.
This blog has documented those views many times, so I will not go over them one by one here. If you want more information, I recommend the film, Maafa21 for documentation on Sanger's beliefs.
Thanks to an upload on YouTube by Planned Parenthood Gala attendee B. Rae Perryman, you can see just how excited Dr. Willie Parker was in accepting the Margaret Sanger award from Planned Parenthood:
Parker has a history with the abortion giant having once worked for them as medical director for Planned Parenthood Metropolitan Washington.
He is fast becoming the Black face Planned Parenthood uses to lessen the effects of those who have documented the genocidal affects of abortion as well as the racist history.
Just as their founder Margaret Sanger vowed to use "Black Ministers" in her "Negro Project," Planned Parenthood is doing the same with Parker.
During his speech, Parker thanked Planned Parenthood for the Margaret Sanger award and told the crowd that his mother shaped his need for recognition when "you act on behalf of others."
"It is in the spirit of my mother's teaching- that- while I am humbled to receive this award- I see you as simply honoring me for what I am supposed to do," Parker told the Planned Parenthood crowd.
Supposed to do? Kill babies is something you are obligated to do?
The abortionist, who dismembers babies including African American babies in the womb, quoted holocaust survivor, Elie Wiesel who said, "Our lives are not our own – they belong to those who desperately need us."
Newflash abortionist Parker:
The unborn baby desperately needs his/her mother and your knife guarantees they will not have the life granted them by their Creator.
Dr. Willie Parker should be ashamed of himself – but he isn't.
Sadly, like others who betrayed their race or participated in the slaughter humans, Parker is proud.
Racist to not provide abortion to Blacks says doc honored by Planned Parenthood
Posted in Abortionist, Blacks promote abortion, Willie Parker with tags Abortion, abortion doctor, Abortionist, Black Genocide, Christian, Dr Seuss, Eugenics, Klan, Margaret Sanger, Mississippi, Planned Parenthood, Racism, racist, Willie Parker, Woodrow Wilson School on February 23, 2015 by saynsumthn
I recently wrote a blog entitled, "Blacks matter to Planned Parenthood if they push abortion." It details how the introduction of abortion by Planned Parenthood came when they conveniently chose a Black man in their leadership.
Now, a Black abortion doctor is playing the Planned Parenthood patsy by trying to "explain" and dismantle the argument that abortion targets the African American community.
Speaking at the Woodrow Wilson School of Public and International Affairs at Princeton University, abortionist Willie Parker, who once called abortion a "sacred decision" said that not providing abortion is racist!
Parker traveled to Mississippi from Alabama to do abortions at the Jackson Women's Health Organization abortion clinics, the last one remaining open in the state.
"I had a traditional understanding of religion and spirituality as a fundamentalist Protestant Christian with a born again experience at the age of 15. But, what that meant for me is that I took my religious belief and understanding very seriously. And so, because of that, while I had not had any explicit teaching that abortion was wrong, although it was kind of implied given that, in my community there were many teens and single women who were forced to continue pregnancies where they were ready or able to parent or not. I chose to err on the side of not violating my conscience or understanding by not providing abortions event though I never questioned a woman's right to make that decision," Parker tells his audience.
Planned Parenthood recently "honored" the abortionist during Black History Month 2015 even placing abortionist Willie Parker on their list of Dream Keeper "Doers."
Abortionist Willie Parker told his Woodrow Wilson School audience that Mississippi is one of the poorest states in the country and we all know the large Black population there.
Pro-lifers know that abortion targets Black communities and Parker is assisting in that agenda, "Women of color and women living in poverty are two communities that are often synonymous but are not always the same tend to have higher rates of unintended pregnancy…"
"That this is the reality befalling Mississippi led me to conclude that the women of Mississippi are amongst the most vulnerable in this country. And hence they are in dire need and very much deserving of my help," the abortionist boasted.
Abortionist Parker who described himself as "a self described protector of human rights" claims that if he could not make women of color a priority for abortion access, who will?
Parker then goes on to attack those who defend the black unborn child, especially those who are members of the African American pro-life community.
"Now don't misunderstand me," he said, "This commitment is not a form of subtle tribalism, because I understand all-too-well as Zora Neale Hurston once said, All my skin folk ain't my kinfolk."
"And, what I mean by that is that some of the most ardent opponents who stand in solidarity with those who oppose abortion are, in my opinion, misguided people of color who also vilify women who make the decision to have abortion, and who exaggerate and expound on the fact that those women are poor and women of color. And, they are the ones who also co-sign, what I call the Black Genocide campaigns," Willie Parker stated.
I guess Parker is also referring to the early members of the civil rights community who also pointed out the genocidal aspects of abortion and birth control as I document here.
Leaders like, Fannie Lou Hamer, who co-founded the Mississippi Freedom Democratic Party and challenged the all white Mississippi delegation for seating at the 1964 Democrat convention in Atlanta City
Hamer once stated, "Once Black women were bought as slaves because they were good breeders. Now they talk about birth control and abortion for blacks. If they'd been talking that way when my mother was bearing children, I wouldn't be here now".
But…I digress…
According to the latest figures by the CDC almost 56% of all abortions reported for race were done on minority women. Of the 405,994 abortions even reported for race or ethnicity in that latest report, 150,942 (37.2%) were performed on White women, 146,856 or (36.2%) were reported on Black women. Hispanic women accounted for 79,408 or (19.7%) while 28,363 or (7.0%) of abortions were categorized as other.
Even abortionist Willie Parker cannot dispute the facts as he says himself:
"One of the things I want to say to kind of demonstrate to you how pressing that issue [Black Genocide] is, when I started in Mississippi, as noble as I thought my efforts were, not everyone was happy to see me."
"What allows [pro-life people] to make that allegation [abortion is a tool of Black Genocide] is a misrepresentation of the very real and concrete epidemiological evidence that shows that disproportionately on a proportionate basis the highest rate of abortion occurs to the Black women followed by Latino women…"
Parker goes on to say that White women have the highest number of abortions, "because there are more white women then there are women of color," he says.
But the real motivation behind Willie Parker's attempt to disprove the abortion is Black Genocide argument is that it is having an affect within the African American community.
"While the legal battle over abortion is occurring at the state and local level as they try and set the stage for overturning Roe – the actual cultural battle for the hearts and minds of the public, is being fought in these ideological wars and framing abortion in ways that are totally misrepresentative over what the issue of abortion is about."
"In this case they've decided as a very effective and cynical strategy to frame abortion as Black Genocide. And that campaign has gained traction over the years. There have been major African American Communities where billboards have been purchased to declare that the most dangerous place for a black baby is in the womb of it's mother…describing Black babies as an endangered species as if they are something other than human in that- as if mothers are totally detached from the pregnancies that they carry."
Parker claims that, "Abortion opponents allege that abortion is a genocidal plot by the government, fronted by Planned Parenthood to build abortion clinics in Black communities and to coerce Black women to use contraception and to have abortionists to kill off black babies…to decimate the Black race. They allege that there is a Negro plot that was put into place by the founder of Planned Parenthood – Margaret Sanger and that this is allegedly core to Planned Parenthood's mission today."
"Let's say I concede all of these arguments are true," Willie Parker says, "What does any of that have to do, in 2015, with a woman, no matter what her color is, making the decision to end a pregnancy, that she does not want?"
WILLIE PARKER DISMISSES MARGARET SANGER'S RACISM
Parker then dismisses the well documented racism pf Planned Parenthood founder, Margaret Sanger, an admitted Klan speaker. This is what Sanger wrote in her autobiography, "I accepted an invitation to talk to the women's branch of the Ku Klux Klan…I saw through the door dim figures parading with banners and illuminated crosses…I was escorted to the platform, was introduced, and began to speak…In the end, through simple illustrations I believed I had accomplished my purpose. A dozen invitations to speak to similar groups were proffered." (Margaret Sanger: An Autobiography, P.366)
Parker, who profits off abortion, works with Planned Parenthood, and has received honors from them, thinks the Black Genocide argument against abortion is ridiculous, "And, with regard to Margaret Sanger's involvement with the Black Community? Margaret Sanger uh uh started birth control clinics in the African American community in Harlem at the invitation of black leaders of her day." he said.
Willie Parker completely ignores the documented plot devised behind the scenes by Sanger and her financier, Clarence Gamble, both active in the Eugenics Society to include blacks in their own demise.
This blog has documented this many times, just do a simple search under "eugenics", "Margaret Sanger" or "Clarence Gamble."
Willie Parker claimed that Sanger's message would benefit African American women. I wonder if that was the focus of her speech to the Klan- how her ideas would benefit Blacks- really Willie are you that naive?
"And. so there was no targeted messaging for the Black community. In fact," says Willie Parker, "Margaret Sanger in keeping with the rest of her day, her attention was to the White community," he says.
Can you believe such ignorance? Sanger's own words make a liar out of Dr. Willie Parker.
As late as 1950 Sanger who openly advocated eugenic sterilization wrote this in a personal letter to Katharine Dexter McCormick, an heir to the International Harvester fortune who used her immense wealth to fund the development of the birth-control pill.
Sanger wrote, "I consider that the world and almost our civilization for the next twenty-five years, is going to depend upon a simple, cheap, safe contraceptive to be used in poverty stricken slums, jungles, and among the most ignorant people. Even this will not be sufficient, because I believe that now, immediately; there should be national sterilization for certain dysgenic types of our population who are being encouraged to breed and would die out were the government not feeding them."
Years earlier, in 1939, Sanger described the American Birth Control League's Negro Project in a letter to fellow eugenicist, Clarence Gamble, "The minister's work is also important and also he should be trained, perhaps by the Federation as to our ideals and the goal that we hope to reach. We do not want the word to go out that we want to exterminate the Negro population and the minister is the man who can straighten out that idea if it ever occurs to any of their more rebellions members."
Parker then calls the Black Genocide argument "noise" and "misleading" and "racial insulting."
Then, Parker shockingly states that not providing abortion to Black women is the real racism:
"I usually reserve terms like brilliant for a positive thing but it is a dastardly jedi-mind trick," he said.
"Because it exploits the notion that we, as a society are very uncomfortable with notions of race.And so, you take people who are well-meaning and want to help people who have the greatest need, in this case, disproportionate women of color.And,so the majority of the healthcare system is still White people, mostly white men, although that's changing with the number of women. So, it really becomes racist NOT to provide this care." abortionist Willie Parker said.
The abortionist, who slaughters children in the womb, ended his speech by sharing the thoughts he says he visits on a regular basis, "In order for this society to grow great," he said, "collectively, we must continue to plant trees under which we will never see it sit. What that means for me is – we have to collectively and on an individual basis do things that will not directly benefit us personally – but that they're the right things to do to make this world the kind of place that we would want to live in."
Abortionist Willie Parker says that on a pretty consistent basis, he will look into the mirror and repeat that exaltation by, "that wise American sage, Dr. Seuss when he says in the Lorax that unless someone like you cares a whole awful lot, Nothing is going to get better. It's not."
May I remind the abortionists that Dr. Suess is a children's favorite – like- the ones he kills in the womb.
Abortionist Willie Parker, who claims abortion is sacred and that not offering it to Black women is racist is not a historian and has little to no knowledge of the eugenics movement, Margaret Sanger, or Planned Parenthood's history. Sadly, Willie Parker is merely one of many Black mouthpieces the abortion lobby has paid off to help eliminate their own race.
If you want a clear and well documented history of abortion and the way it targets the black community- watch the powerful film- Maafa21.
Black pastor: secret board rooms of American corporations know abortion kills future customers
Posted in Black Babies, Black Church, Black Conservative, Black Pastor with tags Abortion, Abortion clinic, Beyonce, Black Babies, customer, Dr. David Manning, Essence, Jay Z, Mc Donald's, Mississippi, Nike, Racism on October 27, 2014 by saynsumthn
A controversial Black pastor from New York has indicated that leaders of Black Pop Culture recognize they are losing customers particularly Black customers because of the high rates of abortion in that community.
Speaking at a rally to protest the last remaining abortion clinic in Mississippi, Dr. David Manning, pastor of the ATLAH World Missionary Church in Harlem, told the crowd a parable, which began by describing a meeting he attended which included corporate leaders as well as pop stars Jay Z and Beyonce.
A parable is not a true story- but a story that portrays a truth.
"I need to let you know that shortly after I began to talk about coming here to Jackson, to the one and only abortion death camp here, in the state of Mississippi," Pastor Manning told the group.
"I received a message from the officer of the Jay Z musical conglomerate organization stating that they wanted to meet with me over this matter of abortion," he explained.
"I came to the meeting. The meeting was at the Four Seasons Hotel in New York City…In that meeting there were a large number of people…Of course, Jay Z and Beyonce, the vice president of sales and marketing for McDonald's Corporation, the vice president and publisher of Essence Magazine, Julian Bond was there, Al Sharpton, Charles Rangel, the president of sales and marketing for Nike, General Motors, several people were there…"
"They began to explain to me these facts: #1. Jay Z began to point out that ten years ago, when he would produce a record, that the record would become immediately popular and they would have a tour afterwards just to celebrate the success of that record. But today, they produce and then they have to go out on the road and spend fifty weeks a year promoting on the road any production of music that they put out there…What they pointed out was this fact and all of them that were there pointed out this fact. That, the population to which they'd distribute their music is dying. And their market share is dying."
Manning wet on, "Ten years ago, there were several million more young people, purchasing their music boosting their economic base and their power. But now they see themselves losing their market share – losing customers – because of one unmitigated fact in America with Black people and Black people in particular. That is, there's not as many births of young people to come along and be customers…And this was the chorus of everything and every person that spoke…They are concerned that because of these death camps- because of these abortion clinics…that the population isn't growing…They all recognize that Black people are dying out as a population…They're concern is this – if abortion continues at the rate it continues – you're going to see a large number of very prominent people and prominent businesses go out of business because they'll have no customers to market to."
Pastor Manning concluded by admitting that, "My invitation to the Four Seasons Hotel is a parable. I just told you a parable. But in their secret board rooms that's what they're talking about – they just don't invite me to come to hear it. But, it's the truth."