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Biomedical subjects

S C Ng

Publications and source records attributed to S C Ng.

At least 127 records · Page 7Linked to original sources
PubMed1997

Sperm head decondensation, pronuclear formation, cleavage and embryonic development following intracytoplasmic injection of mitochondria-damaged sperm in mammals.

The objective of this study was to investigate the influence of sperm mitochondrial destruction on sperm head decondensation, male pronuclear formation, cleavage and embryonic development. In the study two models were used: heterologous (hamster ICSI assay: human sperm injected into a hamster oocyte) for evaluation of sperm head decondensation and pronuclear formation, and homologous (mouse model) for the study of fertilisation and development. Destruction of mitochondria of the sperm was achieved by exposure to cyanide, a respiratory poison. Rhodamine 123 was used to evaluate the functional integrity of mitochondria. Sperm head decondensation was found to be not statistically significantly affected by mitochondrial damage (p = 0.8), with 62.8% and 67.9% condensation in the experimental and control groups respectively. Male pronucleus formation was seen in 40.2% and 44.4% of the injected oocytes in the experimental and control groups respectively. In the mouse experiments 45.5% and 49.7% of the injected oocytes were fertilised in the mitochondria-damaged and live-intact sperm groups respectively (p = 0.53). Development to blastocyst was achieved in 53.5% and 59.4% of the experimental and control groups respectively; the difference was not significant (p = 0.71). Inner cell mass (ICM) cell number were 15.7 +/- 4.12 and 43.1 +/- 11.3 respectively in the mitochondria-damaged group; the equivalent numbers were 14.12 +/- 4.02 and 39.3 +/- 12.6 in the control group. However, the differences in ICM and total cell counts between these two groups were not significant. Of the blastocysts transferred to pseudopregnant mice, 51.3% (20/36) implanted and 33.4% (12/36) developed to live fetuses in the mitochondria-damaged group. These rates were 60.5% (23/38) and 39.5% (15/38) in the control group. In conclusion, this study shows that functional integrity of the sperm mitochondria is not necessary in the process of fertilisation and development when the sperm is deposited into the ooplasm. Fertilisation and development can be achieved by injection of sperm at the very early stage of necrosis in which only the mitochondria have been destroyed and the rest of the cell including the plasma membrane is still intact.

Animals↗
PubMed1997

Is the acrosome reaction a prerequisite for sperm incorporation after intra-cytoplasmic sperm injection (ICSI)?

There is debate as to whether the acrosome reaction is necessary for sperm incorporation after intra-cytoplasmic sperm injection (ICSI). Ultrastructural evidence is presented to show that the acrosome reaction could occur in the ooplasm before sperm incorporation in mature human oocytes or the acrosome could be discarded intact before sperm incorporation in immature oocytes, matured in vitro. Both germinal vesicle and growing follicular oocytes showed sperm chromatin decondensation, with discarded acrosomes close to the sites of incorporation, and were able to form male pronuclei. This is probably the first report of microfertilization of a growing oocyte with a reticulate nucleolus by ICSI. The acrosome reaction, when it occurs, is preceded by acrosome swelling and is followed by vesiculation of surface membranes exposing the inner acrosome membrane, as observed on the surface of the zona during IVF or in the perivitelline space after subzonal sperm injection. These sperm were probably capacitated at the time of ICSI. There was subtle evidence of leaching of the acrosomal matrix from intact discarded acrosomes and from partially depleted acrosomes attached to decondensing spermheads. These sperm were probably not fully capacitated at the time of ICSI. It is concluded that both the acrosome reaction and acrosome deletion are possible prerequisites to sperm incorporation after ICSI.

Acrosome↗
PubMed1997

Fertilization and development of mouse oocytes injected with membrane-damaged spermatozoa.

The objective of this study was to investigate the influence of sperm plasma membrane on fertilization and development in a mouse model. The sperm plasma membrane was destroyed by exposure to Triton X-100 prior to intracytoplasmic sperm injection (ICSI). A single sperm curling (SSC) test was used to evaluate cell viability. The fertilization rates of oocytes obtained following ICSI of membrane-damaged sperm was not significantly higher than that of the control group (62.4 versus 59.6%). Rates of development to blastocyst were also not significantly different (51.7 and 50%). Inner cell mass (ICM) and total embryo cell numbers in the two groups were not statistically different (16 +/- 3.7 versus 14.73 +/- 3.35 and 45.8 +/- 12.5 versus 39.5 +/- 12 respectively). There were no differences in the implantation and live fetus rates between the two groups after transfer to pseudopregnant mice (61.5 and 35.9% respectively for the membrane-damaged group and 53.5 and 31.4% for the intact group). In conclusion, the present study clearly shows that destruction of the sperm plasma membrane does not affect fertilization and further development following injection of membrane-damaged spermatozoa into mouse oocytes. Fertilization and development can be achieved by dead spermatozoa at an early stage of necrosis when only the plasma membrane has been damaged.

Animals↗
PubMed1997

Ongoing normal pregnancy after transfer of zona-free blastocysts: implications for embryo transfer in the human.

Recent reports suggest that transfer of day 5 blastocysts improves implantation rates in in-vitro fertilization programmes. This paper reports a successful ongoing pregnancy after the transfer of zona-free day 6 expanded and hatching blastocysts. The patient was 37 years old and had undergone six stimulated and two thaw cycles previously, all of which had failed. Stimulation was by down-regulation and oocytes recovered transvaginally by ultrasound guidance. Two pronuclear embryos were co-cultured on Vero cells to day 6. The zonae of two hatching and two fully expanded blastocysts were removed using 0.5% pronase, and the zona-free blastocysts were then transferred. Pregnancy was confirmed on day 18 with a positive human chorionic gonadotrophin (HCG) and ultrasound at 6 weeks showed a single healthy fetal heart inside a clear sac. At 14 weeks a triple test (oestriol, J-HCG and alpha-fetoprotein) was normal and at 22 weeks a detailed ultrasound scan showed no congenital anomalies. This is the first report in the human of a normal ongoing pregnancy after the transfer of zona-free day 6 embryos.

Adult↗
PubMed1997

Induction of spermatogenesis in isolated hypogonadotrophic hypogonadism with gonadotrophins and early intervention with intracytoplasmic sperm injection.

Idiopathic hypogonadotrophic hypogonadism (IHH) is a potentially correctable cause of male infertility. However hormonal treatment is usually a slow process and artificial reproductive techniques such as intracytoplasmic sperm injection (ICSI) might be resorted to before full testicular response has been achieved. We report here an unusual variant of IHH of post-pubertal onset in which early intervention with ICSI was attempted. Our patient was 37 years old and presented with male infertility due to azoospermia and undetectable serum gonadotrophin concentrations. He had an apparently normal pubertal development, a testicular volume of 8 ml, normal pituitary-thyroid and pituitary-adrenal function, as well as normal computerized tomographic appearance of the sella region. A combination of human chorionic gonadotrophin (HCG) and menopausal gonadotrophins (HMG) was administered. Spermatozoa were first detected in the semen after 3 months and reached a concentration of approximately 2x10(6)/ml after 9 months. ICSI was attempted at this point; the spermatozoa had good fertilizing ability and three embryos were obtained and replaced. Unfortunately no pregnancy resulted. Treatment with 5000 IU HCG and 150 IU HMG three times per week was continued and sperm counts rose rapidly thereafter to reach 28.3x10(6)/ml after 16 months of injections. His wife conceived naturally during this period and the pregnancy is now in the second trimester. This case illustrates the good prognosis of the rare patient with IHH of post-pubertal onset when treated with gonadotrophins, and suggests that ICSI procedures should be delayed until final testicular maturation is attained.

Adult↗
PubMed1997

Proteus syndrome.

This female Asian (Malay) baby had clinical features of Proteus syndrome. She had a large right facial lipolymphangioma with hyperpigmentation of the overlying skin. There was a smaller lymphangioma over the left side of her neck with excess nuchal folds, macrodactyly and bilateral talipes equinovarus. Despite the extensive hemifacial swelling, there was no evidence of upper respiratory tract obstruction. Generalized seizures developed on the sixth day of life which were controlled with phenobarbital. The lymphangiomas were excised without recurrence.

Female↗
PubMed1997

Chronic low-dose follicle-stimulating hormone compared with clomiphene/human menopausal gonadotropin for induction of ovulation.

Concurrent administration of clomiphene can reduce the amount of gonadotropins required for induction of ovulation. Recently, follicle-stimulating hormone (FSH) administered in a chronic, low-dose fashion has been reported to give satisfactory pregnancy rates. We compared the conventional clomiphene/human menopausal gonadotropin (hMG) with the chronic, low-dose FSH regimen for induction of ovulation in 87 patients over 110 cycles. The clomiphene/hMG regimen required half the amount of gonadotropin compared to the chronic FSH regimen to achieve follicular maturation. Despite the reduced amount of gonadotropin, the clomiphene/hMG regimen induced a mean fourfold higher level of estradiol production and was associated with significantly greater numbers of large and intermediate-sized follicles compared to the chronic FSH regimen. The proportion of clomiphene/hMG cases with multifollicular development and overstimulation was therefore high (30%). In contrast, the chronic FSH regimen, despite requiring larger amounts of gonadotropin and longer periods of treatment, resulted in unifollicular development, low rates of overstimulation and improved pregnancy rates. We conclude that although clomiphene can reduce the requirement for gonadotropins, the relative safety and effectiveness of the chronic low-dose FSH regimen makes it the method of choice for ovulation induction.

Adult↗
PubMed1997

Systemic lupus erythematosus: initial manifestations and clinical features after 10 years of disease.

We studied the initial manifestations and late features in our lupus patients. The clinical data of patients fulfilling the American College of Rheumatology criteria for systemic lupus erythematosus (SLE) were entered prospectively for newly diagnosed patients and cumulatively for those with at least 10 years of disease duration. Ninety-seven Group A (newly diagnosed; 86 females and 11 males; mean age 31 years; 83 Chinese, 11 Malays, and 3 Indians) and 58 Group B (more than 10 years disease duration; 56 females and 2 males; mean age 41 years; 50 Chinese, 5 Malays, and 3 Indians) lupus patients were studied. The commonest clinical features in Group A were: haematological (73%), arthritis (57%), malar rash (43%), renal disorder (31%) and photosensitivity (30%). Group B patients had haematological (78%), malar rash (73%), arthritis (69%), renal disorder (59%) and photosensitivity (33%). Renal disorder was significantly increased over the years (P < 0.001). Hypertension was present in 18% (Group A) and 59% (Group B) (P < 0.00001), diabetes mellitus in 5% (Group A) and 10% (Group B) (P = ns), atherosclerosis in 2% (Group A) and 7% (Group B) (P = ns). Cataract formation was not present in Group A patients but was present in 10% of Group B patients. Renal disorders and morbidity factors like hypertension and cataracts increased significantly over the years. Optimum treatment of lupus patients should therefore include close attention to these factors.

Adolescent↗
PubMed1997

Effects of feeding egg yolk on the serum lipid levels in rabbits.

Healthy, nonobese, young rabbits developed hypercholesterolemia following daily intake of fresh egg yolk for 8 weeks. On a quantitative basis, this diet had a profound effect on serum cholesterol level which rose to 15 or 30 times the baseline value depending on whether the test group consumed the yolk content of one or two eggs during the study. Similar drastic changes observed in low density lipoprotein cholesterol levels in these test animals make this a useful method for defining the early pathological changes of atherosclerosis in man. In addition, the present study provides direct evidence of the deleterious effect of hen's egg yolk on the serum cholesterol level in an appropriate and easily reproducible animal model.

Animals↗
PubMed1997

Endothelin receptor in benign prostatic hyperplastic cells. Binding and functional studies.

Endothelins (ETs) are 21-amino acid peptides that bind to membrane receptors to initiate pathophysiological effects. This report characterizes ET receptors in benign prostatic hyperplasia-1 (BPH-1) cells, a prostate cell line isolated from a specimen of a 60-yr-old man with benign prostatic hyperplasia. [(125)I]ET-1 or -3 binding was of high affinity, with B(max) and K(d) values of 48 fmol/1 x 10(6) cells and 0.16 nM for ET-1, and 2.9 fmol/1 x 10(6) cells and 0.033 nM for ET-3, respectively. ET-1, ET-3, FR139317, Ro 46-2005, and IRL1620 inhibited [(125)I]ET-1 binding to these cells with IC50 values of 0.22, 186, 0.20, 52.8, and 772.3 nM, respectively. Reverse transcription-polymerase chain reaction confirmed that BPH-1 cells expressed more ET(A) than ET(B) receptors. ET-1 did not have any effect on arachidonic acid release, but caused a modest stimulation of phosphatidylinositol hydrolysis, and induced a prominent, sustained elevation in intracellular Ca2+ concentrations. The functional effects of ET-1 were completely inhibited by the ET(A)-selective antagonists FR139317 and A-127722, suggesting that the effects were mediated by the ET(A) receptor. These results suggest that ET may play functional roles in benign prostatic hyperplasia.

Arachidonic Acid↗
PubMed1996

Induction of apoptosis in prostatic tumor cell line DU145 by staurosporine, a potent inhibitor of protein kinases.

We are interested in studying the possibility of modulating prostatic cell growth by manipulating apoptosis. Here we show that 1 microM staurosporine (STS) induces a human androgen-independent prostatic tumor cell line, DU145, to undergo dramatic changes in morphology and results in programmed cell death. Several genes involved in apoptosis were analyzed for expression in STS-treated and untreated DU145 cells. It was observed that these genes were differentially regulated. The expression level of bcl-2, bcl-xL, Ich-1L remains unchanged in treated and untreated cells. On the other hand, DAD1 and interleukin-1 beta-converting enzyme (ICE) were downregulated while bcl-xs and Ich-1s were upregulated. By blocking bcl-2 gene expression using antisense oligonucleotides, it was determined that the anti-bcl-2 oligonucleotides have no effect on the proliferation of DU145 or STS-treated DU145 cells. These results demonstrate that programmed cell death can be induced in an androgen-independent prostatic cancer cell line and BCL-2 was found not to play an important role in preventing STS-induced apoptosis in the DU145 cell line.

Alkaloids↗
PubMed1996

Intracytoplasmic injection of human sperm into the hamster oocyte (hamster ICSI assay) as a test for fertilizing capacity of the severe male-factor sperm.

OBJECTIVE: Our objective was to investigate the fertilizing ability of human sperm from severe male-factor patients, by microinjection of single sperm into the hamster oocyte. DESIGN: Semen samples of severe male factor either with a 0% penetration rate in the zona-free hamster test or with a very low number of motile sperm for which performing the standard penetration test was impossible were used. For the control study, oligozoospermic semen samples with at least 10% penetration rate in zona free hamster test were used. SETTING: All materials were collected from the National University Hospital, Singapore. METHODS: There were 10 patients in both the experimental and the control groups. Intracytoplasmic sperm injection (ICSI) was carried out. The main outcome measures were sperm head decondensation and pronuclear formation. RESULTS: Twenty-one percent of the injected sperm could decondense and undergo male pronuclear formation. This rate was not significantly different from that in the control study group (28%; P = 0.13). A small proportion of the oocytes was damaged during the procedure (9.2 and 8.75% in experimental and control groups, respectively). CONCLUSIONS: Hamster-ICSI assay may be of benefit in predicting the sperm's ability for further development before allowing the patient to undergo the clinical program.

Animals↗
PubMed1996

The sperm centriole: its inheritance, replication and perpetuation in early human embryos.

The inheritance, replication and perpetuation of the sperm centriole in the early human embryo are reported. Both normal monospermic and abnormal dispermic embryos (n = 127) were examined by transmission electron microscopy. Centrioles were traced from fertilization to the hatching blastocyst stage. The sperm proximal centriole is introduced into the oocyte at fertilization and remains attached to the expanding spermhead during sperm nuclear decondensation, as it forms the male pronucleus. A sperm aster is initially formed after the centriole duplicates at the pronuclear stage. At syngamy, centrioles occupy a pivotal position on opposite spindle poles, when the first mitotic figure is formed. Bipolar spindles were found in the majority of embryos, while tripolar spindles were seen in four dispermic embryos at syngamy. Two single centrioles were detected at two poles of two tripolar spindles, while two additional centrioles were located on the sides of a bipolar spindle of a dispermic embryo. Sperm tails were detected near spindle poles at syngamy and in later embryos. Typical centrioles showing the characteristic pin-wheel organization of nine triplets of microtubules were evident. During centriolar replication, the daughter centriole grows laterally from the parent and gradually acquires pericentriolar material (PCM). The two centrioles are surrounded by a halo of electron-dense PCM, which nucleates microtubules, thus making it a typical centrosome. The usual alignment of diplosomes at right angles to each other was maintained. Centrioles were detected at all stages of embryonic cleavage from the 1-cell through 8-cell stages, right up to the hatching blastocyst stage. They were closely associated with nuclei at interphase, when they were often replicating, and were prominently located at spindle poles during the first four cell cycles. In blastocysts, they were detected in trophoblast, embryoblast and endoderm cells respectively. It is evident that the sperm centrosome is the functional active centrosome in human, while the female is inactive but may contribute some centrosomal material to the zygote centrosome. It is very likely that the paternal centriole is the ancestor of the centrioles in fetal and adult somatic cells.

Blastocyst↗
PubMed1996

Fertilization, embryonic development and implantation of mouse oocytes with one or two laser-drilled holes in the zona, and inseminated at different sperm concentrations.

The zonae pellucidae of mouse oocytes were photo-ablated by an ultraviolet laser (337.1 nm) to create one or two 10 microm holes. The pulse energy used was approximately 3 microJ/s, with a frequency of 10 pulse/s. These laser zona-drilled (LZD) oocytes with one hole (LZD1) or two holes (LZD2) and the zona-intact controls were inseminated with spermatozoa at standard concentrations of 5x10(4), 5x10(5), 1x10(6) and 2x10(6)/ml. Fertilization was significantly improved at all sperm concentrations in LZD1 and LZD2 oocytes as compared to the controls. However, there was no significant difference in fertilization rates between LZD1 and LZD2. LZD2 produced significantly higher numbers of blastocysts at day 5. Hatching was significantly enhanced in the presence of either one or two holes in the zona. Polyploidy was generally absent, except in LZD2 oocytes (1%) inseminated at higher sperm concentrations. Differential cell counts of expanded LZD blastocysts were similar to those of the controls. Significantly fewer LZD2 blastocysts implanted and produced viable fetuses than LZD1 and control blastocysts. Morphological abnormalities of the fetuses were absent in all three groups. Laser zona-drilling using the ultraviolet laser was shown to be fast, efficient and safe.

Animals↗
PubMed1996

Fetal and neonatal haemodilution associated with multiple placental chorioangioma: case report.

A pregnancy with polyhydramnios and abnormal antepartum fetal heart rate pattern was found to have multiple placental haemangiomas. Multiple placental haemangiomas can give rise to fetal cardiac failure due to a hyperdynamic circulation or fetal anaemia either due to haemodilution or possibly destruction of blood cells in the chorioangioma. Whether fluid restriction with or without diuretics or blood transfusion is the correct form of treatment of neonatal cardiac failure in such a case is discussed.

Adult↗