BACKGROUND: Although postmenopausal estrogen replacement therapy is known to reduce cardiovascular mortality, the mechanism is not clear yet. Furthermore, the effect of estrogen on vascular tonus is reportedly variable according to the animal models, vascular beds and agonists used. MATERIALS AND METHOD: Bilateral ovariectomies were performed in 12 week-old, 18 spontaneously hypertensive rats (SHR) and 18 normotensive Wistar-Kyoto rats (WKY). Rats were divided into three groups according to the dose of 17beta-estradiol (E 2 ) pellets implanted subcutaneously two weeks after ovariectomy: control (no implantation), low-dose (0.5 mg) and high-dose (5 mg) E 2 replacement group. Two weeks after pellet implantation, organ bath experiments were performed using descending thoracic aortae. For endothelium-dependent relaxation, acetylcholine (10(-9) -3x10(-6) M) was cumulatively added into the vessels precontracted with 10(-7) M norepinephrine (NE). For vasoconstrictor responses, cumulative concentration-contraction curves were constructed in quiescent vessels using NE (10(-9) -10(-5) M), U46619 (10(-9) -3x10(-6) M), endothelin-1 (10(-10) -10(-7) M). In addition, contraction to angiotensin II (10(-7) M) was also obtained. Serum 17beta-estradiol levels were measured by radioimmunoassay. Blood pressure was measured by tail-cuff method in some SHRs before ovariectomy and after placebo/E 2 replacement. RESULTS: Endothelium-dependent relaxation to acetylcholine was impaired in WKY treated with 5 mg E 2 (pIC 50 : control vs 5mg E 2 : 7.75+/-0.13 vs 7.27+/-0.16: n=6: p<0.05). No significant effect was noted in SHR. Contraction to angiotensin II was inhibited by low-dose E 2 in WKY and high-dose E 2 in SHR (% of the contraction to 60 mM KCl: WKY: control vs 0.5 mg E 2 : 39+/-5 vs 25+/-2: SHR: control vs 5 mg E 2 : 34+/-4 vs 22+/-2: n=6 and p<0.05 in WKY and SHR). In contrast, NE-induced contraction was enhanced by E 2 replacement (both low- and high-dose) in WKY and SHR (WKY: control vs 0.5 mg E 2 vs 5 mg E 2 : AUC: 280+/-24 vs 387+/-26 vs 374+/-25: maximal contraction: 137+/-8 vs 166+/-8 vs 162+/-3: pD 2 : 7.63+/-0.11 vs 8.17+/-0.13 vs 8.13+/-0.13: SHR: control vs 0.5 mg E 2 vs 5 mg E 2 : AUC: 265+/-17 vs 349+/-16 vs 406+/-19: maximal contraction: 152+/-6 vs 181+/-9 vs 203+/-16: pD 2 : 7.45+/-0.13 vs 7.91+/-0.08 vs 8.04+/-0.04: n=6 and p<0.05 between control and treated groups in WKY and SHR for all parameters). Contraction to U46619 was enhanced by E 2 replacement in SHR (control vs 0.5 mg E 2 : AUC: 478+/-30 vs 574+/-23: maximal contraction: 181+/-9 vs 230+/-10: n=6: p<0.05 for both parameters). Maximal contractile response to endothelin-1 was also enhanced in SHR (control vs 0.5 mg E 2 vs 5 mg E 2 : maximal contraction: 165+/-7 vs 189+/-7 vs 199+/-8: n=6 and p<0.05 between control and treated groups) but not in WKY. Blood pressure was not different between placebo and E 2- treated SHR (171+/-2 vs 174+/-4 mmHg). CONCLUSION: In WKY, chronic high-dose estrogen replacement impairs endothelium-dependent relaxation to acetylcholine.: low-dose estrogen replacement does not affect endothelium-dependent relaxation in SHR and WKY. Estrogen replacement enhances the contraction to most of the contractile agonists tested except angiotensin II in both WKY and SHR. These results suggest that estrogen replacement affect the vascular tonus differently according to the vasoactive substances and/or hormones without significant effect on blood pressure.
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid ; Acetylcholine ; Angiotensin II ; Animals ; Aorta, Thoracic ; Area Under Curve ; Baths ; Blood Pressure ; Endothelin-1 ; Estrogen Replacement Therapy* ; Estrogens* ; Female ; Models, Animal ; Mortality ; Norepinephrine ; Ovariectomy ; Radioimmunoassay ; Rats ; Rats, Inbred SHR* ; Relaxation

We report a case of tibial nerve palsy after pneumatic tourniquet application for 40 minutes with a tourniquet pressure of 300 mmHg. A 45 years old woman with morbid obesity and diabetes mellitus was underwent partial lateral meniscectomy of left knee. Even 3 months after the event, nerve palsy was not completely recovered. The case underscores the necessity of being aware of the potential for complications associated with tourniquets, despite following recommended guidelines of tourniquet time and pressure. Especially, in the patients with metabolic diseases such as diabetes mellites or obesity, safe duration of tourniquet application may be shortened.
Diabetes Mellitus ; Female ; Humans ; Knee ; Metabolic Diseases ; Obesity ; Obesity, Morbid ; Paralysis ; Tibial Nerve ; Tourniquets

OBJECTIVES: To investigate the effect of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on expression of matrix metalloproteinase-2, 9 (MMP-2, 9) mRNA in mouse embryos. Materials and METHOD: From October 1997 to December 1998, morula stage mouse embryos were cultured for 48 hours with G-CSF and GM-CSF at concentrations of 0.1 pg/ml, 1 pg/ml, 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml, respectively. Embryos not treated with G-CSF or GM-CSF were served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2, 9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p< 0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of all G-CSF treatment groups were significantly increased than in the control, especially in 10, 100 pg/ml and 1 ng/ml treatment groups. The relative quantities of MMP-2 mRNA in all GM-CSF treatment groups were also significantly increased than in the control, especially in 100 pg/ml treatment group. The relative quantities of MMP-9 mRNA of all GM-CSF treatment groups except 10 ng/ml group were significantly increased than in the control, especially 10, 100 pg/ml and 1 ng/ml treatment group. However, the relative quantity of MMP-9 mRNA was significantly increased in only 10 ng/ml G-CSF treatment group than in the control and other treatment groups. CONCLUSION: This study suggests that G-CSF and GM-CSF may increase the m-RNA expression of MMP-2 or 9 in mouse blastocysts with the concentration-specific manner.
Animals ; Blastocyst ; Colony-Stimulating Factors* ; Digestion ; DNA, Complementary ; Embryonic Structures* ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Colony-Stimulating Factor* ; Granulocytes* ; Matrix Metalloproteinase 2* ; Mice* ; Morula ; Reverse Transcription ; RNA, Messenger ; Sensitivity and Specificity ; Sequence Analysis

OBJECTIVES: To examine the in vitro interactions of blastocyst attachment using type I collagen. MATERIALS AND METHODS: ICR mice were used and follicular growth was stimulated by pregnant mare serum gonadotropin and human chorionic gonadotropin. On day 4 of pregnancy, the uteri were removed and blastocysts were flushed. Mixtures of 1mL sterile water, 0.5mL DMEM, 2mL type collagen solution and 0.5mL 0.1M NaOH were prepared and transferred to an incubator where the collagen solution polymerized. Blastocysts were transferred to dishes previously coated with type I collagen. CMRL 1066 was used as the basic culture medium. It was supplemented with 1mM glutamine and 1mM sodium pyruvate plus 50 IU/ml penicillin and 50 mg/ml streptomycin. During the first 4 days the culture medium was supplemented with 20% fetal calf serum and thereafter with 20% heat inactivated human cord serum. All blastocysts were initially cultured for 2 days without media change. After 2 days, fresh medium was renewed daily. The stages of embryo growth were examined and recorded everyday under a dissecting microscope and classified according to the standard in vivo criteria set forth by Witschi. RESULTS: By 48h, nearly all blastocysts had attached to the surface of collagen pad. Following adhesion to the collagen pad, the blastocysts maintained their 3-dimensional integrity in contrast to control. The embryos in collagen pad were not flattening and kept polarity and spherical shape during culture. The polar trophoblast invaded the type I collagen downward unlike the horizontal growth in control. In the developmental stage of mouse blastocyst, there were significant differences between control and type I collagen group during day 4 and 5 culture. CONCLUSION: Blastocyst development was better in type I collagen group than control. Therefore, in vitro culture study using type I collagen could provide improved model for the establishment of blastocyst implantation study.
Animals ; Blastocyst ; Chorionic Gonadotropin ; Collagen ; Collagen Type I* ; Embryo Implantation ; Embryonic Structures* ; Female ; Glutamine ; Gonadotropins ; Hot Temperature ; Humans ; Incubators ; Mice* ; Mice, Inbred ICR ; Penicillins ; Polymers ; Pregnancy ; Pyruvic Acid ; Sodium ; Streptomycin ; Trophoblasts ; Uterus ; Water

It was reported that the etiologies of recurrent spontaneous abortion are immunologic factors, endocrinologic problems, anatomical abnormalities, genetic abnormalities, infection, and unexplained factors. Among those etiologic factors, genetic abnormalities occur in about 5% of the couples who experience recurrent spontaneous abortions, and most common parental chromosomal abnormality contributing to recurrent abortion is balanced translocation. The advent of in vitro fertilization (IVF), the development of skills associated with the handling of human embryo, and an explosion of knowledge in molecular biology have opened the possibility of early diagnosis of genetic disease in preimplantation embryos. Therefore preimplantation genetic diagnosis (PGD) is indicated for couples, infertile or not, at risk of transmitting a genetic disease. A case of successful pregnancy and term delivery by PGD using fluorescence in situ hybridization (FISH) technique in patient with recurrent spontaneous abortion due to balanced translocation is presented with brief review of literatures.
Abortion, Habitual ; Abortion, Spontaneous* ; Blastocyst ; Chromosome Aberrations ; Early Diagnosis ; Embryonic Structures ; Explosions ; Family Characteristics ; Female ; Fertilization in Vitro ; Fluorescence ; Humans ; Immunologic Factors ; In Situ Hybridization ; Molecular Biology ; Parents ; Pregnancy* ; Preimplantation Diagnosis* ; Prostaglandins D

Amylase-producing multiple myeloma is a rare disorder and has poor prognosis. Its characteristics are elevation of salivary type amylase, extensive extramedullary spread, extensive bone destruction, shorter survival time, and abnormal karyotype. Recently, we have experienced a case of amylase-producing IgG kappa type multiple myeloma in a 63-year-old woman. On admission, serum and urine amylase was 8,450U/L and 169,710U/L, 85% and 86% of which was determined to be the salivary type, respectively. The other cause of hyperamylasemia had not been detected. The presence of immunoglobulin and amylase in the myeloma cells was demonstrated immunohistochemically. Bone marrow aspiration smear revealed 59.1% plasma cells. The cytogenetic study of bone marrow cell showed normal karyotype. The patient died 3 months after chemotherapy with melphalan and prednisolone.
Abnormal Karyotype ; Amylases ; Bone Marrow ; Bone Marrow Cells ; Cytogenetics ; Drug Therapy ; Female ; Humans ; Hyperamylasemia ; Immunoglobulin G* ; Immunoglobulins ; Karyotype ; Melphalan ; Middle Aged ; Multiple Myeloma* ; Plasma Cells ; Prednisolone ; Prognosis

This study was performed to investigate the influence of epidermal growth factor (EGF) on preimplantation development, implantation, and expression of epidermal growth factor receptor (EGFR) in mouse embryos. Riverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the presence of transcripts. Following reverse transcription, strategically designed nested primers, optimised for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Eight-cell stage mouse embryos were cultured for 48hrs with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/ml. Embryos not treated with EGF were served as control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 48hrs were determined. The percentages of fully expanded murine blastocysts at 48hrs in all EGF treated group were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in EGF treatment group at 0.1ng/ml (90.7%), 10 ng/ml (89.3%) compared to the control (82.1%; p < 0.05, p < 0.05). The percentages of implanted blastocyst in vitro were significantly higher following incubation with EGF at concentrations of O.lng/ml (38.1%; p < 0.05), 1.0ng/ml (33.3%; p < 0.05), 10ng/ml (22.2%; p < 0.05) compared to the control (10.7%). Embryo development and implantation in vitro were not significantly inhibited or enhanced in cultures supplemented with 100ng/ml EGF compared to the control. The mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF were significantly higher than those of the control and other EGF treatment groups. The implantation rate and mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF group were significantly higher than those of other treatd groups. In conclusion, EGF may have a stimulatory role in embryonic development, implantation and expression of EGFR in embryo itself with concentration-specific manner. These results suggest that EGF may act directly on the mouse embryo and favor its implantaion, irtespective of the presence ar absence of the endometrium.
Animals ; Blastocyst ; Digestion ; DNA, Complementary ; Embryonic Development ; Embryonic Structures* ; Endometrium ; Epidermal Growth Factor* ; Female ; Mice* ; Pregnancy ; Receptor, Epidermal Growth Factor ; Reverse Transcription ; Rivers ; RNA, Messenger ; Sensitivity and Specificity ; Sequence Analysis

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol (E2) were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with 10microliter of 10% DMSO was induced spontaneously in 10.1+/-9.8%, and acrosomal reaction with calcium ionophore A 23187 was induced in 27.4+/-18.1%, and the ARIC value was 17.4+/-16.2%. There were no significant correlation between the ARIC value and the fertilization rate (r2=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos (r2=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the microassisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.
Acrosome Reaction* ; Acrosome* ; Adult ; Calcimycin ; Calcium ; Dimethyl Sulfoxide ; Embryo Transfer ; Embryonic Structures ; Estradiol ; Family Characteristics ; Female ; Fertilization ; Fertilization in Vitro* ; Follicle Stimulating Hormone ; Gonadotropin-Releasing Hormone ; Humans ; Infertility ; Insemination ; Male ; Oocytes ; Ovum ; Pregnancy ; Semen Analysis ; Spermatozoa

Localized fibrous tumor of the scrotum is a very rare disease, and few radiologic features have been reported. We report the sonographic and CT findings of a case of localized fibrous tumor, which developed in the scrotum of a thirty-years-old man.
Mesothelioma* ; Rare Diseases ; Scrotum* ; Ultrasonography

In our previous study, we demonstrated that immobilization stress blocked estrogen-induced luteinizing hormone(LH) surge possibly by inhibiting the synthesis and release of gonadotropin-releasing hormone (GnRH) at the hypothalamic level and by blocking estrogen-induced prolactin (PRL) surge by increasing the synthesis of dopamine receptor at the pituitary level in ovariectomized rats. The present study was performed to determine whether immobilization stress affects pituitary LH responsiveness to GnRH, and whether endogenous opioid peptide (EOP) and dopamine systems are involved in blocking LH and PRL surges during immobilization stress. Immobilization stress was found to inhibit basal LH release and to completely abolish LH surge. However, the intravenous application of GnRH agonist completely restored immobilization-blocked LH surge and basal LH release. Treatment with naloxone did not exert any effect on immobilization-blocked LH surge but increased basal LH release during immobilization stress. Pimozide did not affect immobilization-blocked LH surge or basal LH release. Naloxone also decreased immobilization-induced basal PRL release, but had no effect on immobilization-blocked PRL surge. Immobilization-increased basal PRL levels were augmented by pimozide treatment and immobilization-blocked PRL surge was dramatically restored by pimozide. We conclude that immobilization stress does not impair pituitary LH response to GnRH, and that the immobilization stress-induced blockage of LH surge is probably not mediated by either the opioidergic or the dopaminergic system. However, immobilization-blockade of PRL surge may be partly mediated by the dopaminergic system.
Animal ; Estradiol/*pharmacology ; Female ; Gonadorelin/*pharmacology ; Immobilization ; Luteinizing Hormone/*secretion ; Naloxone/pharmacology ; Opioid Peptides/physiology ; Ovariectomy ; Prolactin/*secretion ; Rats ; Rats, Sprague-Dawley ; Receptors, Dopamine/physiology ; Stress/*metabolism