ABSTRACT
AIM: Minimally invasive surgery (MIS) has been introduced as an alternative to more radical surgical procedures. The Japan Society of Gynecologic and Obstetric Endoscopy and Minimally Invasive Therapy conducted a cross-sectional questionnaire survey to ascertain the status of MIS for endometrial cancer. METHODS: The survey was conducted between May 10 and June 30, 2022. The questionnaire included information on personal attributes, academic affiliations, qualifications, hysterectomies, and intraoperative procedures performed. RESULTS: The total number of questionnaire respondents was 436 (9.2% of the membership). The hysterectomy methods and percentage performed were as follows: simple total hysterectomy (equivalent to benign surgery), 3%; simple total hysterectomy with care to avoid shaving the cervix, 31%; extended total hysterectomy, 48%; and modified radical hysterectomy, 15%. An analysis of hysterectomies performed using MIS for endometrial cancer by qualified gynecologists of endoscopy or board-certified gynecologic oncologists showed a tendency not to choose simple total hysterectomy compared to the gynecologists who did not hold certification (p = 0.019, p = 0.045, and p = 0.010, respectively). Additionally, 67% of respondents did not use uterine manipulators, and 59% of the respondents did not perform lymph node dissection following the guidelines for treating endometrial cancer in Japan. CONCLUSION: This study provided the current status of MIS for endometrial cancer in Japan. The hysterectomy method, use of uterine manipulators, and criteria for omitting lymph node dissection were generally in agreement with the guidelines. Currently, an extra-fascial simple hysterectomy, including at least not shaving the cervix, was a major method for early invasive endometrial cancer using MIS.
Subject(s)
Endometrial Neoplasms , Laparoscopy , Female , Humans , Cross-Sectional Studies , Japan , Endometrial Neoplasms/surgery , Endometrial Neoplasms/pathology , Hysterectomy/methods , Surveys and Questionnaires , Minimally Invasive Surgical Procedures/methods , Retrospective Studies , Laparoscopy/methodsABSTRACT
ConspectusIn this Account, we describe the challenges and promising applications of transmission electron microscopy (TEM) imaging and spectroscopy at cryogenic temperatures. Our work focuses on two areas of application: the delay of electron-beam-induced degradation and following low-temperature phenomena in a continuous and variable temperature range. For the former, we present a study of LiMn1.5Ni0.5O4 lithium ion battery cathode material that undergoes electron beam-induced degradation when studied at room temperature by TEM. Cryogenic imaging reveals the true structure of LiMn1.5Ni0.5O4 nanoparticles in their discharged state. Improved stability under electron beam irradiation was confirmed by following the evolution of the O K-edge fine structure by electron energy-loss spectroscopy. Our results demonstrate that the effect of radiation damage on discharged LiMn1.5Ni0.5O4 was previously underestimated and that atomic-resolution imaging at cryogenic temperature has a potential to be generalized to most of the Li-based materials and beyond. For the latter, we present two studies in the imaging of low-temperature phenomena on the local scale, namely, the evolution of ferroelectric and ferromagnetic domains walls, in BaTiO3 and Y3Fe5O12 systems, respectively, in a continuous and variable temperature range. Continuous imaging of the phase transition in BaTiO3, a prototypical ferroelectric system, from the low-temperature orthorhombic phase continuously up to the centrosymmetric high-temperature phase is shown to be possible inside a TEM. Similarly, the propagation of domain walls in Y3Fe5O12, a magnetic insulator, is studied from â¼120 to â¼400 K and combined with the application of a magnetic field and electrical current pulses to mimic the operando conditions as in domain wall memory and logic devices for information technology. Such studies are promising for studying the pinning of the ferroelectric and magnetic domains versus temperature, spin-polarized current, and externally applied magnetic field to better manipulate the domain walls. The capability of combining operando TEM stimuli such as current, voltage, and/or magnetic field with in situ TEM imaging in a continuous cryogenic temperature range will allow the uncovering of fundamental phenomena on the nanometer scale. These studies were made possible using a MEMS-based TEM holder that allowed an electron-transparent sample to be transferred and electrically contacted on a MEMS chip. The six-contact double-tilt holder allows the alignment of the specimen into its zone axis while simultaneously using four electrical contacts to regulate the temperature and two contacts to apply the electrical stimuli, i.e., operando TEM imaging. This Account leads to the demonstration of (i) the high-resolution imaging and spectroscopy of nanoparticles oriented in the desired [110] zone-axis direction at cryogenic temperatures to mitigate the electron beam degradation, (ii) imaging of low-temperature transitions with accurate and continuous control of the temperature that allowed single-frame observation of the presence of both the orthorhombic and tetragonal phases in the BaTiO3 system, and (iii) magnetic domain wall propagation as a function of temperature, magnetic field, and current pulses (100 ns with a 100 kHz repetition rate) in the Y3Fe5O12 system.
ABSTRACT
Lanternfish, a family Myctophidae, use ventro-lateral body photophores for camouflage of the ventral silhouette, a strategy called counterillumination. While other deep-sea fishes possess pigmented filters and silver reflectors to match sunlight filtering down through the depths, myctophids developed a blue-green reflector for this purpose. In this study, we showed in a lanternfish Diaphus watasei that the reflector comprised monolayered iridophores containing multilayered guanine crystals which enable high reflection with light interference colouration. Platelets shape in body photophores is an unique near-regular hexagonal, probably to allow the homogeneity of reflection angle of the luminescence from photocytes. Focus point of the parabola-like reflector is positioned on the photocytes that ensures the light produced from the photocytes is redirected to the ventral direction. In vitro luminescence reaction using purified luciferase and the substrate coelenterazine showed the light emission at λmax 454â¯nm, while reflection spectra of the iridophores exhibit peaks at longer wavelength, which accomplish to alter the luminescence emitted from photocytes to longer wavelength to fit the mesopelagic light environment. Taken together, we revealed multiple mechanistic elaborations in myctophid body photophores to achieve effective control of biochemical luminescence for counterillumination.
Subject(s)
Fishes/physiology , Animals , Biological Mimicry/physiology , Blood Platelets/chemistry , Blood Platelets/physiology , Fishes/anatomy & histology , Guanine/chemistry , Imidazoles/metabolism , Luciferases/metabolism , Luminescence , Pyrazines/metabolism , Spectroscopy, Fourier Transform Infrared , X-Ray DiffractionABSTRACT
INTRODUCTION: Although endometriosis is a benign disease, it shares some features with cancers, such as invasiveness and the potential to metastasize. This study sought to investigate the epithelial-mesenchymal transition status in human endometriotic lesions. MATERIAL AND METHODS: Thirteen endometriosis patients and 10 control women without endometriosis undergoing surgery for benign indications were recruited. We examined the expression of E-cadherin, vimentin, and epithelial-mesenchymal transition-induced transcriptional factors, such as Snail and ZEB1, by immunohistochemistry. We evaluated the expression of each marker in epithelial cells of both endometriotic lesions (ovarian endometrioma, deep infiltrating endometriosis, adenomyosis) and normal endometria. The correlation between ZEB1 expression and serum level of CA125 was also investigated. RESULTS: Immunohistochemical analysis revealed that although E-cadherin, vimentin, and Snail were expressed in epithelia of normal endometria and endometriotic lesions, ZEB1 expression was only expressed in epithelia of endometriotic lesions. Additionally, ZEB1 was most frequently observed in epithelial cells of invasive endometriosis. The endometriosis patients with high serum CA125 level were more likely to have ZEB1-positive lesions. CONCLUSIONS: This is the first observation of ZEB1 expression in epithelial cells of benign disease. The preferential expression of ZEB1 in epithelial cells of endometriotic lesions suggests that these cells may have, at least in part, a higher level of mesenchymal features possibly via ZEB1-driven epithelial-mesenchymal transition than normal endometria and that ZEB1 can be a potential indicator of invasiveness or severity of endometriosis.
Subject(s)
Biomarkers/metabolism , Endometriosis/diagnosis , Zinc Finger E-box-Binding Homeobox 1/metabolism , Adenomyosis/diagnosis , Adenomyosis/metabolism , Adenomyosis/pathology , Adenomyosis/surgery , Adult , Endometriosis/metabolism , Endometriosis/pathology , Endometriosis/surgery , Female , Humans , Immunohistochemistry , Ligaments/pathology , Neoplasm Invasiveness , Ovarian Diseases/diagnosis , Ovarian Diseases/metabolism , Ovarian Diseases/pathology , Ovarian Diseases/surgery , Predictive Value of Tests , Young AdultABSTRACT
Oxidative stress has a bidirectional role in the development and maturation of zygotes and embryos. Reduction-oxidation reactions and regulatory proteins, such as thioredoxin (TRX) and thioredoxin reductase (TRXR), are intimately involved in the regulation of oxidative stress. The aim of this study was to determine the levels of TRX mRNA and protein in ovarian follicles collected from women undergoing in vitro fertilization (IVF) and to assess these levels relative to follicle size, presence of oocytes, and responsiveness to superovulation. Follicular fluid (FF) and/or granulosa cells (GCs) from large and small follicles were collected at the time of ovum pick-up from 42 IVF patients enrolled in this study. We divided the patients into normal and poor responders (NR and PR, respectively) based on the serum estradiol levels on the day of human chorionic gonadotropin (hCG) administration. We also compared the TRX concentration in FF (FF-TRX) between oocyte-containing follicles (Oc+) and empty follicles (Oc-). The transcript levels of TRX, but not TRXR, were significantly higher in GCs derived from follicles collected from NR than PR, as determined by semi-quantitative RT-PCR analysis. In NR, the FF-TRX was significantly higher in Oc+ follicles than in Oc- follicles and also in large Oc+ follicles than in large Oc- follicles. Unlike NR, PR exhibited no positive association with elevated FF-TRX and presence of oocytes. Based on its collective anti-oxidative, cytoprotective, and cytokine-like properties of TRX, TRX is likely to be involved in the optimal growth and maturation of ovarian follicles and responsiveness to hyperstimulation.
Subject(s)
Fertilization in Vitro , Ovarian Follicle/metabolism , Thioredoxins/genetics , Thioredoxins/metabolism , Adult , Antioxidants/metabolism , Case-Control Studies , Female , Follicular Fluid/metabolism , Granulosa Cells/metabolism , Humans , Infertility/genetics , Infertility/metabolism , Infertility/therapy , Ovulation Induction , Oxidation-Reduction , PregnancyABSTRACT
Repeated and dramatic pregnancy-induced uterine enlargement and remodeling throughout reproductive life suggests the existence of uterine smooth muscle stem/progenitor cells. The aim of this study was to isolate and characterize stem/progenitor-like cells from human myometrium through identification of specific surface markers. We here identify CD49f and CD34 as markers to permit selection of the stem/progenitor cell-like population from human myometrium and show that human CD45(-) CD31(-) glycophorin A(-) and CD49f(+) CD34(+) myometrial cells exhibit stem cell-like properties. These include side population phenotypes, an undifferentiated status, high colony-forming ability, multilineage differentiation into smooth muscle cells, osteoblasts, adipocytes, and chondrocytes, and in vivo myometrial tissue reconstitution following xenotransplantation. Furthermore, CD45(-) CD31(-) glycophorin A(-) and CD49f(+) CD34(+) myometrial cells proliferate under hypoxic conditions in vitro and, compared with the untreated nonpregnant myometrium, show greater expansion in the estrogen-treated nonpregnant myometrium and further in the pregnant myometrium in mice upon xenotransplantation. These results suggest that the newly identified myometrial stem/progenitor-like cells influenced by hypoxia and sex steroids may participate in pregnancy-induced uterine enlargement and remodeling, providing novel insights into human myometrial physiology.
Subject(s)
Antigens, CD34/genetics , Antigens, CD34/physiology , Integrin alpha6/genetics , Integrin alpha6/physiology , Myometrium/metabolism , Stem Cells/physiology , Uterus/physiology , Animals , Cell Differentiation , Cell Hypoxia , Cell Lineage/genetics , Female , Glycophorins/biosynthesis , Glycophorins/genetics , Hematopoietic Stem Cells , Humans , Mice , Myometrium/cytology , Platelet Endothelial Cell Adhesion Molecule-1/biosynthesis , Platelet Endothelial Cell Adhesion Molecule-1/genetics , PregnancyABSTRACT
We experimentally investigate the charge-spin interconversion by introducing a lighter impurity, namely nitrogen (N) into Pt by varying the nitrogen gas flow rate,Qfrom 0 to 20%, and studying the Onsager reciprocity of spin Hall effect (SHE) via complementary methods of spin-torque ferromagnetic resonance and spin-pumping inverse SHE measurements, respectively. We notice a reduction in the crystalline nature of Pt upon nitrogen incorporation. We observe the influence of extrinsic side-jump scattering induced SHE by measuring the spin Hall efficiency,θSHin a wide temperature range from 10 to 296 K. This work establishes the incorporation of N in Pt by using sputtering, to enhance the SHE via extrinsic scattering as well as to observe the reciprocal effects of charge-spin interconversion.
ABSTRACT
Nerve growth factor (NGF) has been recently proposed as one of the key factors responsible not only for promotion of nerve fiber growth but also for the onset and maintenance of pain in a variety of diseases. The aim of this study was to investigate the role of NGF in the pelvic pain associated with endometriosis. Tissue and peritoneal fluid samples were collected from 95 women with laparoscopically and histopathologically confirmed endometriosis and 59 control women without endometriosis. Expression levels of NGF mRNA and protein were examined using real-time RT-PCR and immunohistochemistry, respectively. Concentration of NGF in the peritoneal fluid (PF-NGF) was measured using ELISA. The degree of dyspareunia and dysmenorrhea was evaluated using a verbal rating scale. Real-time RT-PCR analysis revealed that NGF mRNA was significantly more abundant in the ovarian endometriomas and peritoneal endometriosis than in the normal control endometrium. Immunohistochemical analyses demonstrated that NGF was prominently expressed and preferentially localized to the glands of the ovarian endometriomas and peritoneal endometriosis, whereas it was only weakly detectable in the normal endometrium. Although PF-NGF was undetectable in some normal subjects and endometriosis patients, elevated PF-NGF in the peritoneal fluid was more frequently observed in endometriosis patients with severe pain than in those with less severe pain. Our results suggest that NGF produced locally in the peritoneal cavity may be involved in the generation of endometriosis-associated pelvic pain.
Subject(s)
Dysmenorrhea/etiology , Dyspareunia/etiology , Endometriosis/complications , Nerve Growth Factor/physiology , Pelvic Pain/etiology , Adult , Ascitic Fluid/chemistry , Endometriosis/physiopathology , Female , Humans , Nerve Growth Factor/biosynthesis , Up-RegulationABSTRACT
AIM: The purpose of this study was to assess the relationship between chronically impaired spermatogenesis induced by exposing mice to doxorubicin (DXR) and expression of the infertility factor c-kit. METHOD: Eight-week-old male Institute for Cancer Research (ICR) mice were intraperitoneally treated with DXR (0.15 mg/kg, DXR group) or saline (0.15 mg/kg, control group) twice weekly for five weeks and were killed 14 weeks after initial exposure. The animals were sacrificed and bilateral testes were removed and weighed. The testes were stored for the mRNA assay and were fixed for immunohistochemistry. Some testicular samples were fixed in 10% formalin for histopathological examination. RESULTS: Testicular weight (67.6 ± 9.7 mg, P < 0.05), sperm motility (18 ± 6.0%, P < 0.05) and the fertilization rate (2-to-16-cell embryos, 5%; P < 0.05) were significantly lower in the DXR group than in the control group. In the DXR group there was severe tissue damage from the spermatogonia onward, and the Sertoli cell ratio was lower in the DXR group than in the control group (38% vs. 9%, P < 0.05). In addition, there was a decrease in c-kit protein expression, and the amount of c-kit messenger ribonucleic acid (mRNA) expression according to a semiquantitative method was also decreased. CONCLUSION: Expression of c-kit in the mice with chronically impaired spermatogenesis induced by long-term, low-dose administration of DXR correlated with the decrease in the number of spermatogonia.
Subject(s)
Infertility, Male/chemically induced , Proto-Oncogene Proteins c-kit/metabolism , Spermatogenesis , Spermatogonia/metabolism , Animals , Antibiotics, Antineoplastic , Disease Models, Animal , Doxorubicin , Fertilization in Vitro , Immunohistochemistry , Infertility, Male/metabolism , Infertility, Male/pathology , Male , Mice , Organ Size , RNA, Messenger/metabolism , Semen Analysis , Testis/pathologyABSTRACT
RATIONALE: Amniotic membrane is known to have the ability to transdifferentiate into multiple organs and is expected to stimulate a reduced immunologic reaction. OBJECTIVE: Determine whether human amniotic membrane-derived mesenchymal cells (hAMCs) can be an ideal allograftable stem cell source for cardiac regenerative medicine. METHODS AND RESULTS: We established hAMCs. After cardiomyogenic induction in vitro, hAMCs beat spontaneously, and the calculated cardiomyogenic transdifferentiation efficiency was 33%. Transplantation of hAMCs 2 weeks after myocardial infarction improved impaired left ventricular fractional shortening measured by echocardiogram (34+/-2% [n=8] to 39+/-2% [n=11]; P<0.05) and decreased myocardial fibrosis area (18+/-1% [n=9] to 13+/-1% [n=10]; P<0.05), significantly. Furthermore hAMCs transplanted into the infarcted myocardium of Wistar rats were transdifferentiated into cardiomyocytes in situ and survived for more than 4 weeks after the transplantation without using any immunosuppressant. Immunologic tolerance was caused by the hAMC-derived HLA-G expression, lack of MHC expression of hAMCs, and activation of FOXP3-positive regulatory T cells. Administration of IL-10 or progesterone, which is known to play an important role in feto-maternal tolerance during pregnancy, markedly increased HLA-G expression in hAMCs in vitro and, surprisingly, also increased cardiomyogenic transdifferentiation efficiency in vitro and in vivo. CONCLUSIONS: Because hAMCs have a high ability to transdifferentiate into cardiomyocytes and to acquire immunologic tolerance in vivo, they can be a promising cellular source for allograftable stem cells for cardiac regenerative medicine.
Subject(s)
Amnion/cytology , Amnion/physiology , Mesenchymal Stem Cell Transplantation/methods , Myocytes, Cardiac/cytology , Transplantation, Heterologous/physiology , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/physiology , Cell Differentiation , Delivery, Obstetric , Echocardiography , Female , Graft Rejection/prevention & control , Heart/physiology , Humans , Infant, Newborn , Male , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/physiology , Mice , Myocytes, Cardiac/physiology , Pregnancy , Rats , Rats, Wistar , Transplantation Tolerance , Ventricular Function, Left/physiologyABSTRACT
Innate mucosal immune responses, including recognition of pathogen-associated molecular patterns through Toll-like receptors, play an important role in preventing infection in the female reproductive tract (FRT). Damaged cells release nucleotides, including ATP and uridine 5'-diphosphoglucose (UDP-glucose), during inflammation and mechanical stress. We show in this report that P2RY14, a membrane receptor for UDP-glucose, is exclusively expressed in the epithelium, but not the stroma, of the FRT in humans and mice. P2RY14 and several proinflammatory cytokines, such as IL-8, are up-regulated in the endometria of patients with pelvic inflammatory disease. UDP-glucose stimulated IL-8 production via P2RY14 in human endometrial epithelial cells but not stromal cells. Furthermore, UDP-glucose enhanced neutrophil chemotaxis in the presence of a human endometrial epithelial cell line in an IL-8-dependent manner. Administration of UDP-glucose into the mouse uterus induced expression of macrophage inflammatory protein-2 and keratinocyte-derived cytokine, two murine chemokines that are functional homologues of IL-8, and augmented endometrial neutrophil recruitment. Reduced expression of P2RY14 by small interfering RNA gene silencing attenuated LPS- or UDP-glucose-induced leukocytosis in the mouse uterus. These results suggest that UDP-glucose and its receptor P2RY14 are key front line players able to trigger innate mucosal immune responses in the FRT bypassing the recognition of pathogen-associated molecular patterns. Our findings would significantly impact the strategic design of therapies to modulate mucosal immunity by targeting P2RY14.
Subject(s)
Genitalia, Female/pathology , Immunity, Innate , Interleukin-8/genetics , Mucous Membrane/immunology , Receptors, Purinergic P2/physiology , Up-Regulation/genetics , Animals , Chemotaxis , Endometrium/metabolism , Endometrium/pathology , Epithelium/metabolism , Female , Genitalia, Female/immunology , Humans , Interleukin-8/biosynthesis , Mice , Neutrophils/physiology , Receptors, Purinergic P2Y , Uridine Diphosphate Glucose/pharmacology , UterusABSTRACT
We present 2 case reports of anti-N-methyl-d-aspartate receptor (NMDAR) encephalitis that was successfully treated via laparoscopic ovarian cystectomy. In both cases, resection of an ovarian teratoma resulted in eventual full recovery. Although adnexectomy has been reported for tumor resection in anti-NMDAR encephalitis, we chose ovarian cystectomy to preserve ovarian function. The efficacy of cystectomy is equivalent to that of adnexectomy. This suggests that ovarian adnexectomy may not be necessary in anti-NMDAR encephalitis with ovarian teratoma.
Subject(s)
Autoantibodies , Encephalitis/immunology , Ovarian Neoplasms/surgery , Receptors, N-Methyl-D-Aspartate/immunology , Teratoma/surgery , Adult , Female , Humans , Laparoscopy , Ovarian Neoplasms/complications , Teratoma/complicationsABSTRACT
Over the course of pregnancy, the human uterus undergoes a 500- to 1,000-fold increase in volume and a 24-fold increase in weight. The uterine smooth muscle layer or myometrium is remodeled, and both cell hypertrophy and hyperplasia are evident. The origin of the new smooth muscle cells, however, is unclear. They may arise from existing smooth muscle cells, or they may be the product of stem cell differentiation. This study describes a subset of myometrial cells isolated from nonpregnant human myometrium that represents the myometrial stem cell population. This was characterized as side population of myometrial cells (myoSP) by a distinct Hoechst dye efflux pattern. In contrast to the main population of myometrial cells (myoMP), myoSP resided in quiescence, underexpressed or lacked myometrial cell markers, and could proliferate and eventually differentiate into mature myometrial cells in vitro only under low oxygen concentration. Although myoMP displayed mature myometrial phenotypes before and after in vitro cultivation, only myoSP, not myoMP, generated functional human myometrial tissues efficiently when transplanted into the uteri of severely immunodeficient mice. Finally, myoSP were multipotent and made to differentiate into osteocytes and adipocytes in vitro under the appropriate differentiation-inducing conditions. Thus, myoSP exhibited phenotypic and functional characteristics of myometrial stem cells. Study of myoSP will improve the understanding of myometrial physiology and the pathogenesis of myometrium-derived diseases such as leiomyoma. myoSP may also represent a novel source of biological material that could be used in the reconstruction of not only the human uterus but also other organs as well.
Subject(s)
Myometrium/cytology , Myometrium/metabolism , Stem Cells/cytology , Stem Cells/metabolism , Adult , Cell Differentiation , Cell Hypoxia , Cell Separation , Female , Gene Expression Regulation , Humans , Middle Aged , PhenotypeABSTRACT
The atomically flat interface of the Y3Fe5O12 (YIG) thin film and the Gd3Ga5O12 (GGG) substrate plays a vital role in obtaining the magnetization dynamics of YIG below and above the anisotropy field. Here, magnetoimpedance (MI) is used to investigate the magnetization dynamics in fully epitaxial 45 nm YIG thin films grown on the GGG (001) substrates using a copper strip coil in the MHz-GHz frequency region. The resistance (R) and reactance (X), which are components of impedance (Z), allow us to probe the absorptive and dispersive components of the dynamic permeability, whereas a conventional spectrometer only measures the field derivative of the power absorbed. The distinct excitation modes arising from the resonance in the uniform and dragged magnetization states of YIG are respectively observed above and below the anisotropy field. The magnetodynamics clearly shows the visible dichotomy between two resonant fields below and above the anisotropy field and its motion as a function of the direction of the applied magnetic field. A low value of a damping factor of â¼4.7 - 6.1 × 10-4 is estimated for uniform excitation mode with an anisotropy field of 65 ± 2 Oe. Investigation of below and above anisotropy field-dependent magnetodynamics in the low-frequency mode can be useful in designing the YIG-based resonators, oscillators, filters, and magnonic devices.
Subject(s)
Calcinosis , Hysterectomy , Female , Humans , Calcinosis/diagnostic imaging , Postoperative Complications/diagnosisABSTRACT
Progesterone induces decidual transformation of estrogen-primed human endometrial stromal cells (hESCs), critical for implantation and maintenance of pregnancy, through activation of many signaling pathways involving protein kinase A and signal transducer and activator of transcription (STAT)-5. We have previously shown that kinase activation of v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (SRC) kinase is closely associated with decidualization and that SRC is indispensable for maximal decidualization in mice. To address whether SRC kinase activity is essential for decidualization in humans, hESCs were infected with adenoviruses carrying enhanced green fluorescent protein alone (Ad-EGFP), a kinase-inactive dominant-negative mutant (Ad-SRC/K295R), or an inactive autophosphorylation site mutant (Ad-SRC/Y416F). The cells were cultured in the presence of estradiol and progesterone (EP) to induce decidualization and subjected to RT-PCR, immunoblot, and ELISA analyses. Ad-EGFP-infected hESCs exhibited decidual transformation and up-regulation of decidualization markers including IGF binding protein 1 and prolactin in response to 12-d treatment with EP. In contrast, hESCs infected with Ad-SRC/K295R remained morphologically fibroblastoid without production of IGF binding protein 1 and prolactin even after EP treatment. Ad-SRC/Y416F displayed similar but less inhibitory effects on decidualization, compared with Ad-SRC/K295R. During decidualization, STAT5 was phosphorylated on tyrosine 694, a well-known SRC phosphorylation site. Phosphorylation was markedly attenuated by Ad-SRC/K295R but not Ad-EGFP. These results indicate that the SRC-STAT5 pathway is essential for decidualization of hESCs.
Subject(s)
Cell Differentiation/physiology , Decidua/pathology , Endometrium/pathology , STAT5 Transcription Factor/metabolism , Stromal Cells/pathology , src-Family Kinases/metabolism , Animals , Cell Differentiation/drug effects , Cells, Cultured , Decidua/drug effects , Decidua/metabolism , Endometrium/drug effects , Endometrium/metabolism , Enzyme Activation/drug effects , Enzyme Activation/physiology , Epidermal Growth Factor/pharmacology , Estrogens/pharmacology , Female , Humans , Insulin-Like Growth Factor Binding Protein 1/metabolism , Mice , Mitogen-Activated Protein Kinase Kinases/metabolism , NIH 3T3 Cells , Phosphorylation/drug effects , Progesterone/pharmacology , Prolactin/metabolism , Signal Transduction/physiology , Stromal Cells/drug effects , Stromal Cells/metabolismABSTRACT
PURPOSE: To prospectively determine if semiquantitative assessment of R2* images and T1-weighted magnetic resonance (MR) images of leiomyomas correlates with the efficacy of gonadotropin-releasing hormone (GnRH) agonist treatment for volume reduction. MATERIALS AND METHODS: Internal review board approval and informed consent were obtained for this study. Twenty women (mean age, 36.3 years) with intramyometrial leiomyomas were enrolled in this study. Single-section double-echo dynamic MR imaging was performed before GnRH agonist administration. T2-weighted images were obtained before and after two or three GnRH agonist injections (1.88 mg leuprorelin acetate). The steepest signal intensity (SI) upslope on T1-weighted images and the area under the curve (AUC) on R2* images were determined by using a 16 x 16-voxel matrix that was placed in the center of a leiomyoma. Pearson correlation analysis was performed to compare the percentage of volume reduction with SI upslope and AUC. Unpaired t test was performed to evaluate the difference between leiomyomas with AUC and SI upslope values that were less than or greater than the mean. RESULTS: Percentage of volume reduction ranged from 6.2% to 51.1%. The mean AUC and mean SI upslope were 39.2 and 9.83% per second, respectively. There was a significant correlation between the AUC and the percentage of volume reduction (r = 0.81, P < .001), although no significant correlation was observed between the SI upslope and the percentage of volume reduction. A significant difference in percentage of volume reduction was observed in leiomyomas by using mean AUC as a cutoff value (P = .003). CONCLUSION: AUC on R2* images correlates with the efficacy of GnRH agonist before initiation of treatment for volume reduction of leiomyoma.
Subject(s)
Image Interpretation, Computer-Assisted/methods , Leiomyoma/diagnosis , Leiomyoma/drug therapy , Leuprolide/administration & dosage , Magnetic Resonance Imaging/methods , Uterine Neoplasms/diagnosis , Uterine Neoplasms/drug therapy , Adult , Algorithms , Antineoplastic Agents, Hormonal/administration & dosage , Female , Gonadotropin-Releasing Hormone/agonists , Humans , Pilot Projects , Prognosis , Reproducibility of Results , Sensitivity and Specificity , Treatment OutcomeABSTRACT
Recently, structural colour formation and light control by accumulated guanine crystals were reported. However, the relationship between light interference by guanine platelets and light intensity in an individual platelet must be examined further. This study presents experimental evidence that the guanine crystal platelets of fishes aid in efficiently controlling the enhancement of light intensity based on light interference between platelets floating in a micro-space. In addition, a magnetic orientation technique enabled us to dynamically modulate the arrangement of platelets floating in water. A group orientation of the platelets under magnetic fields exhibited a distinct enhancement of the light interference between platelets present in the micro-space, and a two-fold enhancement of the reflected light intensity was achieved by comparing two arrangements of magnetically oriented platelets. The developed micro-optic light control method employing tiny platelets floating under aqueous liquid conditions is expected to facilitate the creation of tuneable optical micro-devices, e.g., a micro-'search-light' for individual cell analysis.
Subject(s)
Guanine/chemistry , Magnetic Fields , Photons , Animals , Crystallization , Goldfish , Optics and PhotonicsABSTRACT
Histone deacetylase inhibitors (HDACIs) have recently emerged as promising anticancer drugs to induce cell cycle arrest, cytodifferentiation, and apoptosis. It is suggested, however, that HDACIs promote cell migration and invasion depending on the cell type. We have reported previously that treatment with HDACIs, including trichostatin A and suberoylanilide hydroxamic acid (SAHA) or progesterone in combination with estrogen, can induce cytodifferentiation of endometrial adenocarcinoma Ishikawa cells through up-regulation of glycodelin, a progesterone-induced endometrial glycoprotein. Given the reported role of glycodelin in cell motility and the migration-modulating potential of HDACIs, we investigated using wound healing assay and transwell migration assay whether ovarian steroid hormones, trichostatin A, or SAHA affects cell migration in endometrial cancer cell lines, Ishikawa and RL95-2. Treatment with ovarian steroid hormones, trichostatin A, and SAHA enhanced cell migration together with up-regulation of glycodelin. SAHA-augmented cell migration was almost completely blocked by gene silencing of glycodelin. Furthermore, overexpression of gycodelin alone resulted in increased cell motility in Ishikawa cells. Our results collectively indicate that glycodelin positively regulates cell motility acting as a mediator of HDACI-enhanced endometrial cell migration, suggesting the involvement of glycodelin in the dynamic endometrial gland morphogenesis during menstrual cycle. Our results raise a possibility that the use of HDACIs in the therapy for glycodelin-inducible endometrial and presumably other gynecological cancers may enhance invasion in cases in which the HDACIs fail to exert differentiation-inducing and/or antiproliferative effects.
Subject(s)
Adenocarcinoma/physiopathology , Cell Movement/drug effects , Endometrial Neoplasms/physiopathology , Enzyme Inhibitors/pharmacology , Glycoproteins/metabolism , Histone Deacetylase Inhibitors , Pregnancy Proteins/metabolism , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Female , Fibronectins/metabolism , Glycodelin , Humans , Hydroxamic Acids/pharmacology , Ovary/metabolism , Up-Regulation , VorinostatABSTRACT
Preimplantation development is marked by four major events: the transition of maternal transcripts to zygotic transcripts, compaction, the first lineage differentiation into inner cell mass and trophectoderm, and implantation. The scarcity of the materials of preimplantation embryos, both in size (diameter < 100 microm) and in quantity (only a few to tens of oocytes from each ovulation), has hampered molecular analysis of preimplantation embryos. Recent progress in RNA amplification methods and microarray platforms, including genes unique to preimplantation embryos, allow us to apply global gene expression profiling to the study of preimplantation embryos. Our gene expression profiling during preimplantation development revealed the distinctive patterns of maternal RNA degradation and embryonic gene activation, including two major transient waves of de novo transcription. The first wave corresponds to zygotic genome activation (ZGA). The second wave, mid-preimplantation gene activation (MGA), contributes dramatic morphological changes during late preimplantation development. Further expression profiling of embryos treated with inhibitors of transcription or translation revealed that the translation of maternal RNA is required for the initiation of ZGA, suggesting a cascade of gene activation from maternal RNA/protein sets to ZGA gene sets and thence to MGA gene sets. To date, several reports of microarray experiments using mouse and human preimplantation embryos have been published. The identification of a large number of genes and multiple signaling pathways involved at each developmental stage by such global gene expression profiling accelerates understanding of molecular mechanisms underlining totipotency/pluripotency and programs of early mammalian development.