The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free 76 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at 120hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUTI1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture (93.8+/-3.1, n=35) is significantly higher than that of control and glucose-free group (76.6 +/- 3.8, n=35 and 68.2+/-4.3, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.
Animals ; Blastocyst ; Cell Count ; Coculture Techniques ; Electrophoresis, Agar Gel ; Embryonic Development ; Embryonic Structures* ; Ethidium ; Female ; Glucose Transport Proteins, Facilitative* ; Glucose* ; Glutaral ; Humans ; Metabolism ; Mice* ; Morula ; Pregnancy ; Pyruvic Acid ; RNA ; Vero Cells

No abstract available.

In parentage testing DNA profiles are used to link the alleged father with paternity by matching their patterns. The probative value of a match is often calculated by multiplying together the estimated frequencies with which each particular VNTR or STR pattern occurs in a reference population. When this calculating method applies to the motherless case of paternity disputes, a calculation must usually be based on types determined for the child and the alleged father. In such case, the first consideration is to exclude a man from paternity of a child when the man did not have the child's paternal allele at some loci, or if the paternal allele cannot be determined, when the man had neither of the child s alleles. The second is to evaluate the DNA evidence when a man is not excluded by the paternal allele. This work is to provide theories of paternity analysis with three approach methods for the motherless case, and to evaluate their efficiency compared to the trio case when the man tested is not excluded. Consequently, the motherless case offers lower probability exclusion and questionable cumulative paternity index than those of the trio case as being typed with 14 STR loci(CSF1PO, TH01, TPDX, vWA, D5S818, D13S317, D7S820, D16S539, FGA, D21S11, FES/FPS, F13A1, D18S80, D17S5). Since the motherless case in paternity disputes is less efficient for paternity exclusion of the child, the use of genetic maker systems with the higher value of mean exclusion chance(MEC) and exact levels of the relative probability of paternity must be of importance considered in the analysis of such deficiency cases.
Alleles ; Child ; Dissent and Disputes ; DNA ; Fathers ; Humans ; Paternity*

No abstract available.

SUMMARY: The present study was carried out to evaluate whether the coculture system of human embryos with Vero cells can improve the quality of embryo or overcome the repetitive implantation failures in order to obtain pregnancy. From January to December 1996, a total 202 cases which patients with the problems of repetitive implantation failures (group I) or those with the poor embryonic quality in their previous cycles (group II) was analysed. The quality of cocultured embryo, pregnancy, on-going and implantation rates between coculture and control groups were compared. Of 93 cases in group I, coculture was performed in 34 cases and conventional IVF for the rest. Of 109 cases in group II, 36 for coculture and 73 for conventional IVF. In group I, pregnancy, on-going and implantation rates in coculture group (14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%), respectively) were higher than those of control (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%), respectively). There is significance in the pregnancy and implantation rates (p=0.028 and p=0.015). In group II, pregnancy, on-going and implantation rates in coculture group (8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%), respectively) were higher than those of control (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%), respectively). Like the result of group 1, there is significance in the pregnancy and implantation rates (p=0.028 and p=0.022). Coculture system with Vero cells works well in the groups of the two indications. Although the case of 3 day-coculture was small as 15 cases in group II, 3 day-coculture improved pregnancy rate (4/15 (26.7%)). Therefore, 3 day-coculture with assisted hatching is recommended to the patients with poor embryonic quality. In conclusion, coculture system with Vero cells can be suggested as an effective method which improves pregnancy rate in those who have repetitive implantation failures or whose embryonic quality was poor in their previous cycles.
Coculture Techniques* ; Embryonic Structures ; Humans ; Humans* ; Pregnancy ; Pregnancy Rate ; Prognosis* ; Vero Cells

No abstract available.
Female ; Follicle Stimulating Hormone* ; Injections, Subcutaneous* ; Ovulation* ; Polycystic Ovary Syndrome*

A purpose of present study is to provide basic information evaluating the utility of Magnetic Resonance imaging as a biological marker estimating manganese effects to central nervous system among welders, which is conducted by comparing urinary and blood manganese concentrations and signal intensities of brain MR images between exposed group and non-exposed group, evaluating the objectivity of subjective grading estimated by correlations between Pallidal signal intensity index (P. I) and subjective grades among exposed group, and comparing the difference of signal intensities according to presence of neurologic symptoms, signs and exposure variables among the exposed group. The exposed group is composed of 11 welders complaining severe symptoms or showing neurological signs, and the non-exposed group is composed of 5 patients who admitted a hospital. Urinary manganese concentrations and signal intensities in T1-weighted MR images among exposed group were higher than those of the non-exposed group significantly, which exhibits that increased signal intensities in T1-weighted MR image represent the effect of manganese exposure. P. Is among the exposed group revealed relatively high correlations with subjective grades ( gamma =0.63, p=0.037) , which suggests the objectivity of subjective grade. Signal intensity in globus pallidus was a suitable single variable representing the effect of manganese accumulation in C.N.S system appropriately, which was verified as follows ; Increased signal intensities among the exposed group had the highest frequency and intensity in the globus pallidus, and the P.I. had a relatively high correlation coefficient ( gamma 0.62, p=0.044) with total score of subjective grades. Signal intensity with subjective grading in globus pallidus represented very high correlation gamma =0.97, p=0.00) with total score of subjective grades, and had a similar correlation coefficient with many variables. It is hard to argue that signal intensities are markers representing pathologic change in C.N.S system or can be used as a diagnostic tool for manganese intoxication, because signal intensities had no difference between the exposed group and the non-exposed group according to presence of neurological signs.
Biological Markers ; Brain ; Central Nervous System ; Globus Pallidus ; Humans ; Magnetic Resonance Imaging* ; Male* ; Manganese ; Neurologic Manifestations

The determination of adjustment method of urinary mercury in spot urine is one of the important issues in assessing the health risks of mercury workers. But there have been debates about whether creatinine or other forms of correction for urinary concentration are better in reducing the variation of urinary mercury. We evaluated four adjustment methods-specific gravity, creatinine, log creatinine and excretion rate-by correlation between values adjusted by the four methods and individual exposure levels which were the geometric mean of daily air mercury level for 2 or 5 days, and mercury concentrations in 24 hour urine were also investigated to compare the results of spot urine. The correlation between values of spot urine and mercury exposure level was over 0.8 in all adjustment methods for workers who worked over 1 year. All four adjustment methods for urinary mercury were found to be similar in assessing the exposure, log creatinine and excretion rate method however were not practical to use due to lack of reference values, and variable standard values of specific gravity. And the creatinine adjusted values were more sensitive in low mercury exposure level. We therefore recommend the creatinine adjustment method for adjustment of urinary mercury.
Adult ; Air Pollutants, Occupational/*urine ; Female ; Human ; Male ; Mercury/*urine ; Middle Age ; Risk Assessment ; Time Factors

No abstract available.
Blood Pressure* ; Obesity*

No abstract available.
Cleft Lip* ; Diagnosis* ; Ultrasonography*