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1.
Clin Exp Obstet Gynecol ; 41(1): 83-6, 2014.
Article in English | MEDLINE | ID: mdl-24707692

ABSTRACT

OBJECTIVE: To examine whether conservative treatment with oral contraceptives is effective in the shrinkage of a peritoneal inclusion cyst (PIC). This is a case report of two patients with a PIC that developed after gynecological surgery. CASES: Both cases were suspected of a PIC based on the medical history, laboratory data, and image findings. It was difficult in differentiate a PIC from an ovarian tumor. Surgery was chosen at first. However, PICs in both cases recurred after surgery and were treated with oral contraceptives as a conservative treatment. PICs shrank after the treatment of oral contraceptives in both cases. CONCLUSION: Due to the high rate of recurrence following surgery, conservative treatment is recommended to treat PICs. Hormone therapy using oral contraceptives seems to have some therapeutic benefit for the PICs.


Subject(s)
Contraceptives, Oral, Combined/administration & dosage , Contraceptives, Oral, Hormonal/administration & dosage , Cysts/drug therapy , Ethinyl Estradiol/administration & dosage , Levonorgestrel/administration & dosage , Peritoneal Diseases/drug therapy , Adult , Cysts/diagnosis , Cysts/etiology , Cysts/surgery , Drug Combinations , Female , Gynecologic Surgical Procedures/adverse effects , Humans , Middle Aged , Peritoneal Diseases/diagnosis , Peritoneal Diseases/etiology , Peritoneal Diseases/surgery , Recurrence , Sclerotherapy , Tissue Adhesions/surgery
2.
Clin Exp Obstet Gynecol ; 40(3): 445-7, 2013.
Article in English | MEDLINE | ID: mdl-24283186

ABSTRACT

BACKGROUND: A heterotopic pregnancy (HP) is an extremely rare disease that represents the simultaneous occurrence of two or more implantation sites in the uterus and extrauterus. Early diagnosis of HP is difficult because of the presence of an intrauterine pregnancy (IUP). In most cases, a precise diagnosis was made after symptoms develop through the rupture or bleeding of the ectopic pregnancy (EP). The authors present a case that was successfully diagnosed as an undemonstrative HP. CASE: A 24-year-old multiparous woman became pregnant after taking clomiphene citrate. At ten weeks of pregnancy, an ultrasonography revealed gestational sacs containing fetuses in the uterus and the right adnexal region, respectively. The patient was diagnosed as having a HP and an emergency right tubal resectomy was performed. The IUP progressed normally and the fetus was delivered at 37 weeks of pregnancy. DISCUSSION: Even if a gestational sac can be confirmed in the uterus, a careful ultrasonographic examination should always be considered to determine the presence of a concurrent extrauterine pregnancy.


Subject(s)
Pregnancy, Heterotopic/diagnosis , Female , Humans , Magnetic Resonance Imaging , Pregnancy , Pregnancy, Heterotopic/diagnostic imaging , Rupture , Ultrasonography, Prenatal , Young Adult
3.
Early Pregnancy (Cherry Hill) ; 5(1): 28-9, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11753500

ABSTRACT

In order to elucidate the regulation of human placental growth during pregnancy, we have assessed PCNA expression, apoptosis and Bcl-2 protein expression in placental trophoblasts over the course of pregnancy. PCNA, Bcl-2 protein and Fas antigen expression were examined by the avidin/biotin immunoperoxidase method, while apoptosis was assessed by in situ DNA 3'-end labeling method. Both PCNA expression and apoptotic DNA fragmentation were noted in cytotrophoblasts (C-cells), being most abundant in very early placenta, less abundant in midterm placenta and least abundant in term placenta. In contrast, Bcl-2 protein expression was noted in syncytiotrophoblasts (S-cells), being least abundant in very early placenta, less abundant in midterm placenta and most abundant in term placenta. These results indicate that very early placenta is characterized by highly proliferative activity of C-cells associated with increased occurrence of apoptosis. Since Bcl-2 protein is an apoptosis-inhibiting gene product, the minimal occurrence of apoptosis in term placenta seems likely to be attributable to the increased expression of Bcl-2 protein in S-cell in term placenta. On the other hand, in extravillous trophoblasts on cell columns, both PCNA and Bcl-2 protein expression were pronounced only in the shallower part, while Fas/Fas ligand expression and apoptosis were prominent in the deeper part. Thus, it seems likely that Bcl-2 protein expression also participates in the regulation of extravillous trophoblast apoptosis.


Subject(s)
Apoptosis/physiology , Proliferating Cell Nuclear Antigen/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Trophoblasts/physiology , Apoptosis/genetics , Cell Division/physiology , DNA Fragmentation , Fas Ligand Protein , Female , Humans , Membrane Glycoproteins/metabolism , Pregnancy , Trophoblasts/cytology , fas Receptor/metabolism
4.
Hum Reprod ; 16(10): 2103-8, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11574499

ABSTRACT

BACKGROUND: The levonorgestrel-releasing intrauterine system (LNg-IUS) has been shown to be effective in the management of menorrhagia. In order to evaluate the effects of LNg-IUS on endometrial proliferation and apoptosis, proliferating cell nuclear antigen (PCNA) expression, apoptosis, Fas and Bcl-2 protein expression in the endometrium were determined at the early proliferative phase of the menstrual cycle before and 3 months after LNg-IUS insertion. METHODS: PCNA, Fas and Bcl-2 protein expression were analysed using an avidin-biotin immunoperoxidase method. Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick-end labelling (TUNEL) method. RESULTS: PCNA, immunolocalized both in the nuclei of endometrial glands and stroma was less abundant 3 months after insertion (P < 0.05). Bcl-2 protein, immunolocalized in the cytoplasm of endometrial glands but not in the stroma, became scanty 3 months after insertion. Fas antigen, immunolocalized only in endometrial glands before insertion, became prominent in both endometrial glands and stroma 3 months after insertion. The apoptosis-positive rate of the nuclei in both endometrial glands and stroma was significantly higher 3 months after insertion relative to that before insertion (P < 0.05). CONCLUSIONS: LNg-IUS resulted in a decrease in endometrial proliferation and an increase in apoptosis in endometrial glands and stroma. The increase in apoptosis associated with increased Fas antigen expression and decreased Bcl-2 protein expression in the endometrium may be one of the underlying molecular mechanisms by which LNg-IUS insertion causes the atrophic change of the endometrium.


Subject(s)
Apoptosis/drug effects , Endometrium/drug effects , Endometrium/pathology , Levonorgestrel/administration & dosage , Adult , Cell Division/drug effects , Drug Delivery Systems , Endometriosis/etiology , Endometrium/metabolism , Female , Humans , Levonorgestrel/therapeutic use , Menorrhagia/complications , Menorrhagia/drug therapy , Menorrhagia/metabolism , Menorrhagia/pathology , Proliferating Cell Nuclear Antigen/metabolism , Proto-Oncogene Proteins c-bcl-2/antagonists & inhibitors , Time Factors , fas Receptor/metabolism
5.
Endocr J ; 47(3): 317-27, 2000 Jun.
Article in English | MEDLINE | ID: mdl-11036876

ABSTRACT

In order to evaluate placental trophoblast proliferation and apoptosis during pregnancy, we investigated proliferating cell nuclear antigen (PCNA) expression, apoptosis and Bcl-2 protein expression in the human placenta using avidin/biotin immunoperoxidase method to examine PCNA and Bcl-2 protein expression, and TUNEL method to assess apoptosis. The appearance of apoptotic cells in very early term placental trophoblasts was also examined by transmission electron microscopy. PCNA was immunolocalized in the nuclei of cytotrophoblasts (C-cells). Determination of the mean percentage of PCNA-positive nuclei of C-cells revealed that PCNA expression in C-cells was highest in very early term (4th to 5th wk) placentas and significantly decreased with the advance of pregnancy. Bcl-2 protein was immunolocalized in the cytoplasm of syncytiotrophoblast (S-cell), being least abundant in very early term placentas, less abundant in early term and midterm placentas, and most abundant in term placentas. On the basis of TUNEL method, apoptosis was apparent in the nuclei of both C-cells and S-cell. The apoptosis positive rate of C-cell nuclei was highest in very early term 4th to 5th wk placentas, and significantly decreased in early term 7th to 9th wk and midterm placentas, but somewhat increased in term placentas compared to that in midterm placentas. On the other hand, apoptosis positive rate of S-cell nuclei was remarkably higher only in very early term 4th to 5th wk placentas compared to that in early term, midterm and term placentas. Transmission electron microscopy revealed the appearance of apoptotic nucleus in very early term placental trophoblasts. These results demonstrate for the first time that apoptosis in the human normal placenta predominates in both C-cells and S-cell in very early term 4th to 5th wk pregnancy and drastically diminished after 7th wk of pregnancy. An apparent increase in apoptosis in C-cells in term placentas compared to that in midterm placentas may reflect aging of the placenta or parturition-associated biological change. The abundant expression of Bcl-2 protein in S-cell in term placentas may be responsible for the diminished occurrence of apoptosis in S-cell in term placentas.


Subject(s)
Apoptosis , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Trophoblasts/cytology , Abortion, Induced , Cell Division , Female , Humans , In Situ Nick-End Labeling , Microscopy, Electron , Pregnancy , Proliferating Cell Nuclear Antigen/analysis , Trophoblasts/metabolism , Trophoblasts/ultrastructure
6.
J Neurosci ; 19(5): 1728-35, 1999 Mar 01.
Article in English | MEDLINE | ID: mdl-10024359

ABSTRACT

Molecular cloning studies have revealed the existence of a large family of voltage-gated K+ channel genes expressed in mammalian brain. This molecular diversity underlies the vast repertoire of neuronal K+ channels that regulate action potential conduction and neurotransmitter release and that are essential to the control of neuronal excitability. However, the specific contribution of individual K+ channel gene products to these neuronal K+ currents is poorly understood. We have shown previously, using an antibody, "KC, " specific for the Kv2.1 K+ channel alpha-subunit, the high-level expression of Kv2.1 protein in hippocampal neurons in situ and in culture. Here we show that KC is a potent blocker of K+ currents expressed in cells transfected with the Kv2.1 cDNA, but not of currents expressed in cells transfected with other highly related K+ channel alpha-subunit cDNAs. KC also blocks the majority of the slowly inactivating outward current in cultured hippocampal neurons, although antibodies to two other K+ channel alpha-subunits known to be expressed in these cells did not exhibit blocking effects. In all cases the blocking effects of KC were eliminated by previous incubation with a recombinant fusion protein containing the KC antigenic sequence. Together these studies show that Kv2.1, which is expressed at high levels in most mammalian central neurons, is a major contributor to the delayed rectifier K+ current in hippocampal neurons and that the KC antibody is a powerful tool for the elucidation of the role of the Kv2.1 K+ channel in regulating neuronal excitability.


Subject(s)
Hippocampus/metabolism , Potassium Channels, Voltage-Gated , Potassium Channels/metabolism , Potassium/metabolism , Animals , Antibodies/pharmacology , Antibody Specificity , Cells, Cultured , Delayed Rectifier Potassium Channels , Dendrites/metabolism , Embryo, Mammalian , Fibroblasts/drug effects , Fibroblasts/metabolism , Hippocampus/drug effects , In Vitro Techniques , Mice , Microscopy, Fluorescence , Neurons/drug effects , Neurons/metabolism , Patch-Clamp Techniques , Potassium Channels/drug effects , Potassium Channels/immunology , Rats , Shab Potassium Channels , Transfection
7.
Exp Anim ; 47(4): 229-35, 1998 Oct.
Article in English | MEDLINE | ID: mdl-10067165

ABSTRACT

Morphological and immunohistochemical features of the abdominal mesotheliomas that were developed by inoculation of 3 cell lines (MeET-4, -5 and -6) established from spontaneous abdominal mesotheliomas in male F344 rats. Although the original tumors of three cell lines showed signs of epithelioid growth with a predominantly simple papillary pattern, transplanted tumors revealed a variety of morphologic features including epithelioid with glandular structures, sarcomatous, and a mixture of these components. All tumor cells of transplanted tumors were positive for alpha-smooth muscle actin (ASMA) but almost negative for desmin as were epithelioid cells of the original tumors, and the cell lines were positive for desmin but not for ASMA. These results suggested that mesothelioma in the F344 rat had the potential for wide spectrum differentiation under in vitro conditions. The microenvironmental factors obtained in vivo can modify their potential ability and their morphological aspects. These factors may be related to tumor cell reexpression of ASMA of tumor cells that were masked under in vitro culture conditions.


Subject(s)
Abdominal Neoplasms/pathology , Gene Expression Regulation, Neoplastic , Mesothelioma/pathology , Neoplasm Transplantation , Abdominal Neoplasms/genetics , Abdominal Neoplasms/veterinary , Actins/biosynthesis , Animals , Cell Differentiation , Cell Survival , Immunohistochemistry , Male , Mesothelioma/genetics , Mesothelioma/veterinary , Rats , Rats, Inbred F344 , Tumor Cells, Cultured
8.
Mol Pharmacol ; 52(5): 821-8, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9351973

ABSTRACT

The voltage-gated delayed-rectifier-type K+ channel Kv2.1 is expressed in high-density clusters on the soma and proximal dendrites of mammalian central neurons; thus, dynamic regulation of Kv2.1 would be predicted to have an impact on dendritic excitability. Rat brain Kv2.1 polypeptides are phosphorylated extensively, leading to a dramatically increased molecular mass on sodium dodecyl sulfate gels. Phosphoamino acid analysis of Kv2.1 expressed in transfected cells and labeled in vivo with 32P shows that phosphorylation was restricted to serine residues and that a truncation mutant, DeltaC318, which lacks the last 318 amino acids in the cytoplasmic carboxyl terminus, was phosphorylated to a much lesser degree than was wild-type Kv2.1. Whole-cell patch-clamp studies showed that the voltage-dependence of activation of DeltaC318 was shifted to more negative membrane potentials than Kv2.1 without differences in macroscopic kinetics; however, the differences in the voltage-dependence of activation between Kv2.1 and DeltaC318 were eliminated by in vivo intracellular application of alkaline phosphatase, suggesting that these differences were due to differential phosphorylation. Similar analyses of other truncation and point mutants indicated that the phosphorylation sites responsible for the observed differences in voltage-dependent activation lie between amino acids 667 and 853 near the distal end of the Kv2.1 carboxyl terminus. Together, these parallel biochemical and electrophysiological results provide direct evidence that the voltage-dependent activation of the delayed-rectifier K+ channel Kv2. 1 can be modulated by direct phosphorylation of the channel protein; such modulation of Kv2.1 could dynamically regulate dendritic excitability.


Subject(s)
Potassium Channels, Voltage-Gated , Potassium Channels/physiology , Animals , Brain/metabolism , COS Cells/metabolism , Delayed Rectifier Potassium Channels , Membrane Potentials , Patch-Clamp Techniques , Phosphorylation , Point Mutation , Potassium Channels/chemistry , Potassium Channels/genetics , Potassium Channels/metabolism , Precipitin Tests , Rats , Serine/chemistry , Shab Potassium Channels , Transfection
9.
J Cell Biol ; 135(6 Pt 1): 1619-32, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8978827

ABSTRACT

The voltage-sensitive K+ channel Kv2.1 has a polarized and clustered distribution in neurons. To investigate the basis for this localization, we expressed wild-type Kv2.1 and two COOH-terminal truncation mutants, delta C318 and delta C187, in polarized epithelial MDCK cells. These functional channel proteins had differing subcellular localization, in that while both wild-type Kv2.1 and delta C187 localized to the lateral membrane in high density clusters, delta C318 was expressed uniformly on both apical and lateral membranes. A chimeric protein containing the hemagglutinin protein from influenza virus and the region of Kv2.1 that differentiates the two truncation mutants (amino acids 536-666) was also expressed in MDCK cells, where it was found in high density clusters similar to those observed for Kv2.1. Polarized expression and clustering of Kv2.1 correlates with detergent solubility, suggesting that interaction with the detergent insoluble cytoskeleton may be necessary for proper localization of this channel.


Subject(s)
Potassium Channels, Voltage-Gated , Potassium Channels/metabolism , Animals , Binding Sites , Biotin/metabolism , Brain/cytology , COS Cells , Cell Line , Cell Membrane/metabolism , Cell Polarity , Cytoplasm/metabolism , Delayed Rectifier Potassium Channels , Male , Mutagenesis , Neurons/metabolism , Octoxynol , Potassium Channels/genetics , Rats , Rats, Sprague-Dawley , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Shab Potassium Channels , Solubility
10.
Br J Pharmacol ; 116(3): 2062-6, 1995 Oct.
Article in English | MEDLINE | ID: mdl-8640346

ABSTRACT

1. In the present study we estimated the KA value of endothelin-1 (ET-1) for ETA-receptors by a new method in which the level of expression of ETA-receptors in Xenopus oocytes was altered in a controlled way. 2. Kvl.2 (a delayed rectifier type K channel) c RNA at the fixed concentration of 0.2 micro g micro l(-1) was mixed with ETA-receptor cRNA at various concentration ratios (10(-3)-3). Oocytes were examined 2-4 days after the injection of the cRNA mixtures. 3. In these oocytes, ET-1 suppressed the amplitude of Kvl.2 current in a dose-dependent manner in the range of 0.1-100 nM; the maximum inhibition produced by ET-1 was larger and the EC50 value for the inhibition by ET-1 was smaller as the mixture ratio was increased. Double-reciprocal plots of equiactive concentrations of ET-1 in 1/1- and 1/30-injected oocytes yielded a KA for ET-1 of 7.4 nM. The number of ETA-receptors in 1/30-injected oocytes was 13% of that in 1/1-injected oocytes, whereas the inhibition of the current in 1/30-injected oocytes was about 60% of that in 1/1-injected oocytes. This suggests the presence of spare receptors of ETA in the latter. 4. A saturation binding experiment estimated a KD value of 0.1 nM for ET-1 at ETA-receptors and the number of ETA-receptors in 1/30-injected oocytes was 23% of that in 1/1-injected ones. This value was not significantly different from that estimated by the above new method. However, there was a discrepancy between KA and KD, which could be due to factors unique to the expression system employed in the present study.


Subject(s)
Endothelins/metabolism , Oocytes/metabolism , Receptors, Endothelin/metabolism , Analysis of Variance , Animals , Binding, Competitive , Dose-Response Relationship, Drug , Female , Oocytes/cytology , Patch-Clamp Techniques , RNA, Complementary/genetics , RNA, Complementary/metabolism , Receptor, Endothelin A , Xenopus laevis
11.
Glycoconj J ; 12(3): 290-7, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7496144

ABSTRACT

We previously reported for the first time two Japanese patients with aspartylglycosaminuria (AGU). A novel disialo Asn N-glycoside (AG-5) has been isolated from the urine of one of the patients in addition to four known monosialo Asn N-glycosides (AG-1 to AG-4) by gel filtration and anion exchange chromatography in this study. Final purification of AG-5 was achieved by an electrochemical chromatographic method, high performance liquid chromatography with pulsed amperometric detector (HPLC-PAD). The yield of AG-5 was approximately 1 mg l-1 urine. The chemical structures of AG-1 to AG-5 were characterized by gas-liquid chromatography, a permethylation study, fast atom bombardment-mass spectrometry (FAB-MS), and nuclear magnetic resonance (NMR). Based on the structural analysis, AG-5 had the following novel structure: NeuAc alpha 2-->8NeuAc alpha 2-->3Gal beta 1-->4GlcNAc beta 1-->Asn.


Subject(s)
Acetylglucosamine/analogs & derivatives , Glycosides/urine , Acetylglucosamine/urine , Adult , Carbohydrate Sequence , Chromatography, High Pressure Liquid , Female , Humans , Molecular Sequence Data
12.
J Biol Chem ; 269(39): 24138-42, 1994 Sep 30.
Article in English | MEDLINE | ID: mdl-7929069

ABSTRACT

We constructed tandem cDNA by linking the 5' end of a delayed rectifier-type (Kv1.2) clone to the 3' end of a transient-type (Kv1.4) K+ channel clone. Fusion genes were also constructed, consisting of Kv1.4 and mutants of Kv1.2, which have a single amino acid substitution in the S4-S5 loop. From electrophysiological characterization, it is likely that two pairs of tandem heterodimer constructs can form hybrid channels. In addition, it has been revealed that the wild-type hybrid channel shows a time constant of inactivation very similar to that observed in the homotetrameric Kv1.4 channel. Difference of inactivation kinetics between wild-type and mutant hybrid K+ channels suggests that not only the S4-S5 loop of Kv1.4 but also that of Kv1.2 can serve as the acceptor sites for the inactivation gates, and that all of four sets of loops should be functional for rapid inactivation. From these results, in the hybrid channels the structure and composition of the acceptor sites could be important factors for determining the rate of inactivation.


Subject(s)
Potassium Channels/genetics , Amino Acid Sequence , Animals , DNA, Complementary , Kinetics , Membrane Potentials , Molecular Sequence Data , Potassium Channel Blockers , Potassium Channels/physiology , Recombinant Fusion Proteins , Xenopus
13.
Eur J Pharmacol ; 268(3): 451-4, 1994 Aug 16.
Article in English | MEDLINE | ID: mdl-7805772

ABSTRACT

To investigate mechanisms for the receptor-mediated inhibition of a rat cardiac K+ channel clone (KV1.2), we coexpressed KV1.2 with a subtype of endothelin receptors (ETA) in Xenopus oocytes. Effects of endothelin ETA receptor stimulation were mimicked by application of PMA (4-beta-phorbol 12-myristate 13-acetate; 0.1 microM) or intracellular injection of CaCl2 (estimated concentration of 1 microM). These effects diminished in the presence of staurosporine (1 microM) or EGTA (estimated concentration of 5 mM). These results suggest that both activation of protein kinase C and an increase in intracellular Ca2+ contribute to the suppression.


Subject(s)
Potassium Channels/metabolism , Receptors, Endothelin/metabolism , Alkaloids/pharmacology , Animals , Calcium/metabolism , Myocardium/metabolism , Oocytes/metabolism , Patch-Clamp Techniques , Phosphatidylinositols/metabolism , Plasmids/physiology , Potassium Channels/drug effects , Protein Kinase C/antagonists & inhibitors , RNA, Complementary/biosynthesis , Rats , Receptors, Endothelin/genetics , Second Messenger Systems/physiology , Staurosporine , Tetradecanoylphorbol Acetate/pharmacology , Xenopus
14.
Biochem Biophys Res Commun ; 184(3): 1484-9, 1992 May 15.
Article in English | MEDLINE | ID: mdl-1590805

ABSTRACT

We have cloned a delayed rectifier type K channel from rat heart (RH1). RH1 was identical to the rat brain K channel BK2 and differed from recently cloned rat cardiac K channel RAK by one amino acid residue. Endothelin receptors(ETRs)-mediated modulation of RH1 current (IRH1) was studied using Xenopus oocyte expression system. Activation of two different subtypes of ETRs by endothelin-1 equally suppressed the amplitude of IRH1. Stimulation of phosphatidylinositol turnover will probably be responsible for the suppression.


Subject(s)
Endothelins/pharmacology , Heart/physiology , Potassium Channels/genetics , Potassium Channels/physiology , Animals , Cloning, Molecular , DNA/genetics , Evoked Potentials/drug effects , Female , Genetic Vectors , Male , Membrane Potentials/drug effects , Oocytes/drug effects , Oocytes/physiology , Phosphatidylinositols/metabolism , Potassium Channels/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Recombinant Proteins/drug effects , Recombinant Proteins/metabolism , Tetraethylammonium , Tetraethylammonium Compounds/pharmacology , Xenopus laevis
15.
Jikken Dobutsu ; 36(4): 443-8, 1987 Oct.
Article in Japanese | MEDLINE | ID: mdl-3436378

ABSTRACT

A personal computer system was developed for measuring values of copulatory behavior of small laboratory rodents. This system enabled us to get the analysis data of copulatory behavior of male rats as soon as possible after observation. This present system could be useful to measure other reproductive behavior of small rodents.


Subject(s)
Computers , Copulation , Microcomputers , Rodentia/physiology , Animals , Female , Male , Rats
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