57 RELATIONSHIP BETWEEN FETAL ABNORMALITIES AND PERIPHERAL STEROID CONCENTRATIONS DURING GESTATION IN COWS TRANSFERRED WITH EMBRYOS PRODUCED BY SOMATIC CELL NUCLEAR TRANSFER

2010 ◽  
Vol 22 (1) ◽  
pp. 187
Author(s):  
M. Hirako ◽  
H. Takahashi ◽  
K. Kimura ◽  
N. Adachi ◽  
S. Akagi
Keyword(s):  
Repeated Measures ◽  
Nuclear Transfer ◽  
Well Being ◽  
First Trimester ◽  
Left Ventricular ◽  
Histopathological Analysis ◽  
Efficient Production ◽  
Fetal Abnormality ◽  
Fetal Abnormalities

Cloning of mammals by nuclear transfer frequently results in gestational or neonatal failure with a variety of abnormalities that are likely caused by inappropriate epigenetic reprogramming. Early diagnosis of fetal abnormality is important for efficient production of cloned animals. Sex steroids are produced in the bovine placenta and their levels in the blood might be useful as a measure of fetal well-being, as in humans. The objective of this study was to investigate whether changes in peripheral levels of progesterone and estrogens reflect fetal abnormalities. Donor cells for nuclear transfer were obtained from subculture of cumulus cells retrieved from ovarian follicles of a Japanese Black cow. Recipient oocytes were derived from ovaries obtained at an abattoir and matured in vitro. Metaphase II oocytes were enucleated and each fused with a donor cell by DC pulses. Nuclear transferred oocytes were activated and cultured for 7 days. Embryos developed to the blastocyst stage were transferred into the uterine horn ipsilateral to the ovary bearing the CL of 32 multiparous Japanese Black and Holstein crossbred cows at 7 to 8 days after the day of standing estrus (Day 0). Blood was collected from Day 40 until parturition. Progesterone and estrogens in the blood plasma of 6 recipient cows with full-term delivery were measured by RIA. These profiles were compared with each other and with the changes in a cow made pregnant by MOET. Statistical differences were analyzed with repeated measures ANOVA. Parturition was induced on Day 290. Stillborn, dead, or euthanized calves were subjected to necropsy and histopathological analysis. Pregnant cows were 14, 13, and 9 on Days 30, 60, and 90, respectively. Thereafter, 3 aborted around Days 110, 120, and 190. Six cows delivered calves weighing 45.8 ± 2.1 kg (mean ± SEM) on Days 291 to 293. Their birth weights were greater than those of female calves (31.2 ± 0.4 kg, n = 6) produced by MOET in the same breed. Three calves grew normally until weaning. One was stillborn due to dystocia, but no abnormalities were observed except for large offspring syndrome. One was euthanized 2 days after natural delivery due to ananastasia. Thymic and thyroid hypoplasia, left ventricular dysplasia, and pulmonary fibrosis were found by necropsy. Another calf delivered by Caesarean section died of infirmity 2 days later and had thymic and thyroid hypoplasia. Changes in plasma steroid concentrations were consistent with each other and with those in a MOET cow except the last one, in which progesterone levels tended to be higher during the first trimester and estrogens were lower during the last half of gestation. Progesterone levels tended to be lower in cows bearing a healthy calf than in cows bearing a weak calf before parturition. These results imply that peripheral steroid levels may reflect fetal normality, although large offspring syndrome does not affect their concentrations.

36 CHANGES IN PLASMA CONCENTRATIONS OF PROGESTERONE AND ESTROGENS DURING GESTATION IN COWS WITH STILLBORN SOMATIC CELL CLONED CALVES

2011 ◽  
Vol 23 (1) ◽  
pp. 124
Author(s):  
M. Hirako ◽  
H. Takahashi ◽  
M. Aoki ◽  
H. Ishizaki ◽  
Y. Kariya ◽  
...  
Keyword(s):  
Repeated Measures ◽  
Nuclear Transfer ◽  
Plasma Concentrations ◽  
Well Being ◽  
Late Gestation ◽  
Histopathological Analysis ◽  
Efficient Production ◽  
Fetal Abnormality ◽  
Plasma Progesterone ◽  
Estrone Sulfate

Cloning of mammals by nuclear transfer frequently results in gestational or neonatal failure with a variety of abnormalities that are likely caused by inappropriate epigenetic reprogramming. Early diagnosis of fetal abnormality is important for efficient production of cloned animals. Sex steroids are produced in the bovine placenta and their levels in the blood might be useful as a measure of fetal well being, as they are in humans. The objective of this study was to investigate whether changes in peripheral concentrations of progesterone and estrogens reflect fetal conditions. Donor cells for nuclear transfer were obtained from subculture of cumulus cells retrieved from ovarian follicles of a Japanese Black cow. Recipient oocytes were derived from ovaries obtained at an abattoir and matured in vitro. Metaphase II oocytes were enucleated and each fused with a donor cell by DC pulses. Nuclear transferred oocytes were activated and cultured for 7 days. Embryos developed to the blastocyst stage were transferred into the uterine horn ipsilateral to the ovary bearing the CL of 29 multiparous Japanese Black and Holstein crossbred cows at 7 to 8 days after the day of standing oestrus (Day 0). Blood was collected at regular intervals from Day 40 until parturition. Plasma progesterone, estrone, oestradiol-17β, and estrone sulfate were measured by RIA in 4 recipient cows. 2 vaginally delivered healthy calves weighed 35 and 36 kg on days 278 and 280, respectively. The other 2 delivered stillborn calves weighed 42 and 31 kg by Caesarean section on days 267 and 287, respectively. Steroid profiles were compared with each other and with those in a cow made pregnant by embryo transfer. Statistical differences at stages of gestation were analysed with repeated-measures ANOVA. Stillborn calves were subjected to necropsy and histopathological analysis. Significant differences in steroid levels were observed individually and temporarily. In the cow bearing the former stillborn, progesterone and oestradiol-17β concentrations tended to be lower during mid and late gestation, respectively. Large offspring syndrome, ventricular septal defect, pulmonary oedema, hepatic fibrosis, and placental dysplasia were found in the calf by necropsy and histopathological analysis. In the cow bearing the second stillborn, plasma progesterone concentrations were temporarily increased from days 220 to 230 and reached a peak of 16 ng mL–1, which was approximately 3 times as high as before (5.0 ± 0.9; mean ± s.d.), then followed by a similar transient increase in estrogens during days 240 and 250 (170 pg mL–1, 16 pg mL–1, and 21 ng mL–1 at peak of estrone, oestradiol-17β, and estrone sulfate, respectively). Thereafter, oestrogen concentrations stayed low until parturition. As internal organs of the stillborn calf were severely autolysed with polyhydramnios at birth, the fetus might have been dead around Day 250. These results imply that changes in peripheral steroid concentrations in some cases reflect fetal condition.

scholarly journals Early Detection of Fetal Abnormality

2013 ◽  
Vol 7 (1) ◽  
pp. 46-50
Author(s):  
Ritsuko K Pooh
Keyword(s):  
Early Detection ◽  
High Frequency ◽  
Obstet Gynecol ◽  
Three Dimensional ◽  
First Trimester ◽  
Two Dimensional ◽  
Fetal Abnormality ◽  
Fetal Abnormalities ◽  
Comprehensive Information ◽  
Vaginal Transducer

ABSTRACT After introduction of high-frequency vaginal transducer, transvaginal two-dimensional ultrasound has established a field of sonoembryology and most of the major fetal abnormalities have been detectable in the first trimester. Three-dimensional ultrasound adds an objective and comprehensive information to two-dimensional sonographic findings. How to cite this article Pooh RK. Early Detection of Fetal Abnormality. Donald School J Ultrasound Obstet Gynecol 2013;7(1):46-50.

19 CHANGES IN PLASMA STEROID CONCENTRATIONS DURING GESTATION IN COWS WITH SPONTANEOUS ABORTION OF SOMATIC CELL CLONED FETUSES

2012 ◽  
Vol 24 (1) ◽  
pp. 121
Author(s):  
M. Hirako ◽  
H. Takahashi ◽  
K. Kimura ◽  
N. Adachi ◽  
S. Akagi
Keyword(s):  
Sex Steroids ◽  
Repeated Measures ◽  
Nuclear Transfer ◽  
Plasma Concentrations ◽  
Donor Cell ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Steroid Synthesis ◽  
Estrone Sulfate

Cloning of mammals by nuclear transfer frequently results in gestational failure with a variety of abnormalities that are likely due to inappropriate epigenetic reprogramming. Monitoring the placental function during gestation is important to clarify the cause of abnormalities in cloned animals. Sex steroids are produced in the bovine placenta and their levels in maternal peripheral blood are a useful measure of placentation. The objective of this study was to investigate changes in plasma concentrations of sex steroids during gestation in cows aborting cloned fetuses. Donor cells for nuclear transfer were obtained from subculture of cumulus cells retrieved from ovarian follicles of a Japanese Black cow. Recipient oocytes were derived from ovaries obtained at an abattoir and matured in vitro. Metaphase II oocytes were enucleated and each fused with a donor cell by DC pulses. Nuclear-transferred oocytes were activated and cultured for 7 days. Embryos developed to the blastocyst stage were each transferred into the uterine horn ipsilateral to the ovary bearing the CL of 39 multiparous Japanese Black and Holstein crossbred cows at 7 to 8 days after the day of standing oestrus (day 0). Fourteen recipient cows were diagnosed pregnant on Day 40 by ultrasonography and 7 cows delivered at full term. The other seven miscarried on Day 66, 81, 85, 89, 97, 104 and 211. Blood was collected from these cows at least once a week following the pregnancy diagnosis. Progesterone, estrone, oestradiol-17β and estrone sulfate in the blood plasma were measured by RIA and were compared with those in pregnant AI cows. Statistical differences at stages of gestation were analysed with repeated-measures ANOVA. In all miscarried cows, progesterone concentrations were similar to those in AI cows until several days before abortion and then rapidly decreased to the basal level. Concentrations of all estrogens stayed low until abortion in six cows aborting by day 104, whereas estrone and oestradiol-17β started to increase around Day 80 and estrone sulfate gradually increased from around Day 50 and started to increase drastically around Day 80 in AI cows. In another cow aborting on Day 211, profiles of estrone and oestradiol-17β were similar to those in AI cows until around Day 150. Thereafter, concentrations of these estrogens gradually decreased to the basal levels by Day 160 and stayed low until abortion. In this cow, gradual increase in estrone sulfate during Day 50 to 80 was not observed, but the difference in the concentration was not statistically significant from AI cows. The following profile of estrone sulfate was similar to those in active estrogens. The fetus was still alive on day 160 and fetal death was confirmed on day 180 by ultrasonography. These results suggest the possibility that developmental or functional failure of placenta associated with steroid synthesis may be a cause of mid-term miscarriage of a cloned fetus.

20 IMMUNOCYTOCHEMICAL CHARACTERIZATION OF DAYS 17 AND 19 OVINE IN VIVO AND SOMATIC CELL NUCLEAR TRANSFER EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 110
Author(s):  
N. I. Alexopoulos ◽  
K. Schauser ◽  
M. K. Holland ◽  
T. T. Peura ◽  
K. M. Hartwich ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Primordial Germ Cells ◽  
Well Being ◽  
Research Centre ◽  
Pluripotency Marker ◽  
Β Tubulin

The present study aimed to characterize development of in vivo and somatic cell nuclear transfer (SCNT) Merino ovine embryos on Days 17 and 19 using the mesoderm marker vimentin, the neuroectoderm marker β-tubulin III, and the pluripotency marker OCT4 for primordial germ cells. In vivo embryos were obtained by transferring 10 to 20 zygotes to each of 20 intermediate recipient ewes. On Day 6, 4 final recipients were used to obtain embryos for recovery at Day 17 (n = 2) and at Day 19 (n = 2). SCNT embryos were constructed from in vitro-matured oocytes and adult granulosa cells. On Day 6, 9 embryos (with intact inner cell masses) were transferred to each recipient for collection at Day 17 (n = 2) and Day 19 (n = 2). Ewes were euthanazed with phenobarbitone and excised reproductive tracts flushed with saline. A total of 24 embryos at Day 17 (14 in vivo and 10 SCNT) and 23 embryos at Day 19 (11 in vivo and 12 SCNT) were collected and processed for immunohistochemistry. On Day 17, length of embryo proper for in vivo and SCNT embryos was 6.2 and 5.6 mm, width of allantois was 9.3 and 3.8 mm, and number of somites was 19 and 13, respectively. On Day 19, length of embryo proper for in vivo and SCNT embryos was 11.5 and 7.3 mm, width of allantois was 70.6 and 13.7 mm, and number of somites was 28 and 22, respectively. On both Day 17 and Day 19, in vivo embryos had a much larger allantois, more somites, and were longer in length. Vimentin staining was observed in all in vivo embryos at Day 17 and Day 19; however, in 75% of Day 17 SCNT embryos, differentiation of the somites into dermatome with underlying sclerotome was either delayed or absent. Similarly, at both Day 17 and Day 19, a larger proportion of embryos did not stain for β-tubulin III, and of those that did, only a small amount was found in the neural tube. The intensity of both stains was much weaker in the SCNT embryos compared to in vivo embryos. Oct-4 was initially found in the splanchnic mesoderm lining the endoderm in the wall of the yolk sac and later in the dorsal mesentery and medial aspect of the mesonephros with in vivo embryos having double the number of positively stained cells than SCNT embryos. The delay in mesoderm compartments of SCNT embryos could imply future problems in the musculoskeletal system. However, lack of any neuroectoderm staining in more than half SCNT embryos could point to neurological problems being more of an issue in terms of survival and well-being. Equally, the smaller allantois could induce later placental abnormalities. The current project was funded by the Co-operative Research Centre for Innovative Dairy Products (CRC-IDP), Australia.

scholarly journals Digital Methods to Optimize Pregnant Women’s Participation in Longitudinal Research (Preprint)

2019 ◽  
Author(s):  
Beth McGee ◽  
Marie Leonte ◽  
Kevin Wildenhaus ◽  
Martha Wilcox ◽  
Jenna Reps ◽  
...  
Keyword(s):  
Mental Health ◽  
Pregnant Women ◽  
Repeated Measures ◽  
Longitudinal Research ◽  
Well Being ◽  
First Trimester ◽  
Target Audience ◽  
Data Set ◽  
Digital Methods ◽  
High Degree

BACKGROUND The scientific and medical communities require advanced study of maternal health and well-being to develop improved outcomes for mothers and their babies but can be challenged measuring mental health during pregnancy and into postpartum due to accessibility and cooperation issues. OBJECTIVE This work employs digital methods to describe mental health and maternal experience from the first trimester of pregnancy through 3 months postpartum among a cohort similar to the general population of pregnant women in the US. The objective was to collect a data set of longitudinal measures for epidemiologists to develop predictive modeling to determine early in pregnancy which women will develop perinatal depression with a high degree of accuracy. METHODS The authors recruited 1,179 pregnant women from BabyCenter.com and followed them with repeated measures using custom, mobile optimized, questionnaires administered online. The authors employed a combination of typical and novel digital strategies to minimize attrition and build trust among participants. In all stages of recruitment, data collection, and participant engagement, the authors adapted their strategies to meet the target audience and leveraged digital means to achieve the final goal. RESULTS Active participants completed 7,757 assessments. These assessments contribute to a robust data set that is enabling predictive models that can identify with a high degree of accuracy women at risk of developing mood disorders before symptoms onset. CONCLUSIONS The study’s success at retaining pregnant women through the postpartum period was based on leveraging a platform with an already established reach and trust among the target audience (BabyCenter), and adapting engagement methodologies tailored to the needs of this target audience.

scholarly journals Transcriptomic analysis of primate placentas and novel rhesus trophoblast cell lines informs investigations of human placentation

2020 ◽  
Author(s):  
Jimi L. Rosenkrantz ◽  
Jessica E. Gaffney ◽  
Victoria HJ. Roberts ◽  
Lucia Carbone ◽  
Shawn L. Chavez
Keyword(s):  
Animal Model ◽  
Cell Lines ◽  
Pregnancy Complications ◽  
Rhesus Macaques ◽  
Enrichment Analysis ◽  
Well Being ◽  
First Trimester ◽  
Trophoblast Cell ◽  
Pathway Enrichment Analysis

AbstractProper placentation, including trophoblast differentiation and function, is essential for the health and well-being of both the mother and baby throughout pregnancy. Placental abnormalities that occur during the early stages of development are thought to contribute to pre-eclampsia and other placenta-related pregnancy complications. However, relatively little is known about these stages in humans due to obvious ethical and technical limitations. Rhesus macaques are considered an ideal surrogate for studying human placentation, but the unclear translatability of known human placental markers and lack of accessible rhesus trophoblast cell lines can impede the use of this animal model. Here, we performed a cross-species transcriptomic comparison of human and rhesus placenta and determined that while the majority of known placental markers were similarly expressed, 952 differentially expressed genes (DEGs) were identified between the two species. Pathway enrichment analysis of the 447 human-upregulated DEGs, including ADAM12, ERVW-1, KISS1, LGALS13, PAPPA2, PGF, and SIGLEC6, revealed over-representation of functional terms associated with pre-eclampsia and other pregnancy disorders. Additionally, to enable in vitro functional studies of early placentation, we generated and thoroughly characterized two highly-pure first-trimester telomerase (TERT) immortalized rhesus trophoblast cell lines (iRP-D26 and iRP-D28A) that retained crucial features of isolated primary trophoblasts. Overall, our findings help elucidate the molecular translatability between human and rhesus placenta and reveal notable expression differences in human placental markers and genes associated with pregnancy complications that should be considered when using the rhesus animal model to study normal and pathological human placentation.

scholarly journals Transcriptomic analysis of primate placentas and novel rhesus trophoblast cell lines informs investigations of human placentation

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Jimi L. Rosenkrantz ◽  
Jessica E. Gaffney ◽  
Victoria H. J. Roberts ◽  
Lucia Carbone ◽  
Shawn L. Chavez
Keyword(s):  
Animal Model ◽  
Cell Lines ◽  
Pregnancy Complications ◽  
Rhesus Macaques ◽  
Well Being ◽  
First Trimester ◽  
Trophoblast Cell ◽  
Functional Enrichment ◽  
Marker Genes

Abstract Background Proper placentation, including trophoblast differentiation and function, is essential for the health and well-being of both the mother and baby throughout pregnancy. Placental abnormalities that occur during the early stages of development are thought to contribute to preeclampsia and other placenta-related pregnancy complications. However, relatively little is known about these stages in humans due to obvious ethical and technical limitations. Rhesus macaques are considered an ideal surrogate for studying human placentation, but the unclear translatability of known human placental markers and lack of accessible rhesus trophoblast cell lines can impede the use of this animal model. Results Here, we performed a cross-species transcriptomic comparison of human and rhesus placenta and determined that while the majority of human placental marker genes (HPGs) were similarly expressed, 952 differentially expressed genes (DEGs) were identified between the two species. Functional enrichment analysis of the 447 human-upregulated DEGs, including ADAM12, ERVW-1, KISS1, LGALS13, PAPPA2, PGF, and SIGLEC6, revealed over-representation of genes implicated in preeclampsia and other pregnancy disorders. Additionally, to enable in vitro functional studies of early placentation, we generated and thoroughly characterized two highly pure first trimester telomerase (TERT) immortalized rhesus trophoblast cell lines (iRP-D26 and iRP-D28A) that retained crucial features of isolated primary trophoblasts. Conclusions Overall, our findings help elucidate the molecular translatability between human and rhesus placenta and reveal notable expression differences in several HPGs and genes implicated in pregnancy complications that should be considered when using the rhesus animal model to study normal and pathological human placentation.

Effects of Rest and Exercise on Cardiac Blood Volume Determinations

1997 ◽  
Vol 36 (08) ◽  
pp. 259-264
Author(s):  
N. Topuzović
Keyword(s):  
Radionuclide Ventriculography ◽  
Left Ventricular ◽  
Peak Exercise ◽  
Volume Determination ◽  
Group I ◽  
Lv Volumes ◽  
Group Ii ◽  
Lv Volume

Summary Aim: The purpose of this study was to investigate the changes in blood activity during rest, exercise and recovery, and to assess its influence on left ventricular (LV) volume determination using the count-based method requiring blood sampling. Methods: Forty-four patients underwent rest-stress radionuclide ventriculography; Tc-99m-human serum albumin was used in 13 patients (Group I), red blood cells was labeled using Tc-99m in 17 patients (Group II) in vivo, and in 14 patients (Group III) by modified in vivo/in vitro method. LV volumes were determined by a count-based method using corrected count rate in blood samples obtained during rest, peak exercise and after recovery. Results: In group I at stress, the blood activity decreased by 12.6 ± 5.4%, p <0.05, as compared to the rest level, and increased by 25.1 ± 6.4%, p <0.001, and 12.8 ± 4.5%, p <0.05, above the resting level in group II and III, respectively. This had profound effects on LV volume determinations if only one rest blood aliquot was used: during exercise, the LV volumes significantly decreased by 22.1 ± 9.6%, p <0.05, in group I, whereas in groups II and III it was significantly overestimated by 32.1 ± 10.3%, p <0.001, and 10.7 ± 6.4%, p <0.05, respectively. The changes in blood activity between stress and recovery were not significantly different for any of the groups. Conclusion: The use of only a single blood sample as volume aliquot at rest in rest-stress studies leads to erroneous estimation of cardiac volumes due to significant changes in blood radioactivity during exercise and recovery.

Evaluation of Normal Heart Rate in Early Pregnancy Corresponding to Gestational Age between Six to Eight Weeks

Med Phoenix ◽  
2017 ◽  
Vol 2 (1) ◽  
pp. 34-37
Author(s):  
Akhilesh Kumar Jha ◽  
Bikranta Rimal ◽  
Tarannum Khatun
Keyword(s):  
Heart Rate ◽  
Early Pregnancy ◽  
Well Being ◽  
Clinical Decision ◽  
First Trimester ◽  
Normal Heart ◽  
Normal Heart Rate ◽  
First Trimester Of Pregnancy ◽  
Singleton Pregnancies

Background: Ultrasonography is the reliable and safe way for the evaluation of pregnancy. Heart rate can be detected more confidently from the Ultrasonography. Heart rate is an important parameter for the evaluation of early pregnancy. The purpose of this study was to evaluate the normal heart rate in embryos/fetuses between 6 and 8 weeks of gestation.Method: In our region people are poor and most of them do not know the benefit of regular follow up examination during pregnancy. So most of pregnant women come to our centre at late stage of pregnancy. The number of pregnancy cases is good in our centre but the number of early pregnancy cases coming to regular follow up examination is low. Thus the study was conducted in 51 normal singleton pregnancies undergoing routine ultrasound examination during the first trimester of pregnancy. The duration of study was 6 weeks.Result: Out of 51 singleton pregnancies, 20 cases (39.2%) heart rate were between 131-150 beat per minute and 25 cases (49.0 %) heart rate were between 151-170 beat per minute. However 4 cases (7.8%) were between 110-120 beat per minute and 2 cases (3.9%) were more than 171 beat per minute. There were zero cases above the 180 beat per minute.Conclusion: The result of this study will help to evaluate abnormal and normal fetal heart rate so that early clinical decision whether to continue the pregnancy or terminate it can be taken, as Ultrasonography is only the method used in screening fetal well being in most of the region of our country.Med Phoenix Vol.2(1) July 2017, 34-37

The Uniform Status of Children of Assisted Conception Act: Does It Protect the Best Interests of the Child in a Surrogate Arrangement?

1990 ◽  
Vol 16 (4) ◽  
pp. 525-553
Author(s):  
Mimi Yoon
Keyword(s):  
Artificial Insemination ◽  
Medical Technology ◽  
Well Being ◽  
Best Interests ◽  
Medical Procedures ◽  
Assisted Conception ◽  
Regulatory Scheme ◽  
Infertile Couples ◽  
Definition Of

Medical technology is easing the plight of many infertile couples by offering such reproductive alternatives as in vitro fertilization, artificial insemination and surrogacy. In response to the changes in our society's definition of family, wrought by scientific advances, the National Conference of Commissioners on Uniform States Laws promulgated the Uniform Status of Children of Assisted Conception Act. The purpose of this Act is to protect the interests of children born through extraordinary medical procedures. This Note analyzes the Act's provisions regarding surrogacy and focuses on how the Commission's regulatory scheme fails to protect the child's interests. The Act's alternative of voiding the surrogacy contract also does not protect the child's interests. A more complete regulatory scheme which protects the adult parties’ interests, as well as the child's, should be devised, as the adequacy of the adult parties’ protection ultimately affects the child's well-being.

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