RESUMO
Observational human data and several lines of animal experimental data indicate that maternal obesity impairs offspring health. Here, we comprehensively tested the model that maternal obesity causes defects in the next three generations of oocytes and embryos. We exposed female F0 mice to a high-fat/high-sugar (HF/HS) diet for 6 weeks before conception until weaning. Sires, F1 offspring and all subsequent generations were fed control chow diet. Oocytes from F1, F2 and F3 offspring of obese mothers had lower mitochondrial mass and less ATP and citrate than oocytes from offspring of control mothers. F0 blastocysts from HF/HS-exposed mice, but not F1 and F2 blastocysts, had lower mitochondrial mass and membrane potential, less citrate and ATP and smaller total cell number than F0 blastocysts from control mothers. Finally, supplementation of IVF media with the anti-oxidant mito-esculetin partially prevented the oocyte mitochondrial effects caused by maternal HF/HS diet. Our results support the idea that maternal obesity impairs offspring oocyte quality and suggest that antioxidant supplementation should be tested as a means to improve IVF outcomes for obese women.
Assuntos
Dieta Hiperlipídica/efeitos adversos , Desenvolvimento Embrionário/fisiologia , Obesidade/patologia , Oócitos/crescimento & desenvolvimento , Açúcares/efeitos adversos , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Feminino , Fertilização in vitro , Células HEK293 , Humanos , Exposição Materna , Fenômenos Fisiológicos da Nutrição Materna , Potencial da Membrana Mitocondrial/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Umbeliferonas/farmacologiaRESUMO
STUDY QUESTION: Does supplementation with co-enzyme Q10 (CoQ10) improve the oocyte mitochondrial abnormalities associated with obesity in mice? SUMMARY ANSWER: In an obese mouse model, CoQ10 improves the mitochondrial function of oocytes. WHAT IS KNOWN ALREADY: Obesity impairs oocyte quality. Oocytes from mice fed a high-fat/high-sugar (HF/HS) diet have abnormalities in mitochondrial distribution and function and in meiotic progression. STUDY DESIGN, SIZE, DURATION: Mice were randomly assigned to a normal, chow diet or an isocaloric HF/HS diet for 12 weeks. After 6 weeks on the diet, half of the mice receiving a normal diet and half of the mice receiving a HF/HS diet were randomly assigned to receive CoQ10 supplementation injections for the remaining 6 weeks. PARTICIPANTS/MATERIALS, SETTING, METHODS: Dietary intervention was initiated on C57Bl6 female mice at 4 weeks of age, CoQ10 versus vehicle injections were assigned at 10 weeks, and assays were conducted at 16 weeks of age. Mice were super-ovulated, and oocytes were collected and stained to assess mitochondrial distribution, quantify reactive oxygen species (ROS), assess meiotic spindle formation, and measure metabolites. In vitro fertilization was performed, and blastocyst embryos were transferred into control mice. Oocyte number, fertilization rate, blastulation rate and implantation rate were compared between the four cohorts. Bivariate statistics were performed appropriately. MAIN RESULTS AND THE ROLE OF CHANCE: HF/HS mice weighed significantly more than normal diet mice (29 versus 22 g, P< 0.001). CoQ10 supplementation did not influence weight. Levels of ATP, citrate, and phosphocreatine were lower and ROS levels were higher in HF/HS mice than in controls (P< 0.001). CoQ10 supplementation significantly increased the levels of metabolites and decreased ROS levels in oocytes from normal diet mice but not in oocytes from HF/HS mice. However, CoQ10 completely prevented the mitochondrial distribution abnormalities observed in the HF/HS mice. Overall, CoQ10 supplementation significantly increased the percentage of normal spindle and chromosome alignment (92.3 versus 80.2%, P= 0.039). In the sub-analysis by diet, the difference did not reach statistical significance. When undergoing IVF, there were no statistically significant differences in the number of mature oocytes, the fertilization rate, blastocyst formation rates, implantation rates, resorption rates or litter size between HF/HS mice receiving CoQ10 or vehicle injections. LIMITATIONS, REASONS FOR CAUTION: Experiments were limited to one species and strain of mice. The majority of experiments were performed after ovulation induction, which may not represent natural cycle fertility. WIDER IMPLICATIONS OF THE FINDINGS: Improvement in oocyte mitochondrial distribution and function of normal, chow-fed mice and HF/HS-fed mice demonstrates the importance of CoQ10 and the efficiency of the mitochondrial respiratory chain in oocyte competence. Clinical studies are now needed to evaluate the therapeutic potential of CoQ10 in women's reproductive health. STUDY FUNDING/COMPETING INTERESTS: C.E.B. received support from the National Research Training Program in Reproductive Medicine sponsored by the National Institute of Health (T32 HD040135-13) and the Scientific Advisory Board of Vivere Health. K.H.M received support from the American Diabetes Association and the National Institute of Health (R01 HD083895). There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: This study is not a clinical trial.
Assuntos
Mitocôndrias/efeitos dos fármacos , Obesidade/metabolismo , Oócitos/efeitos dos fármacos , Ubiquinona/análogos & derivados , Animais , Peso Corporal/efeitos dos fármacos , Peso Corporal/fisiologia , Dieta Hiperlipídica , Modelos Animais de Doenças , Feminino , Camundongos , Mitocôndrias/metabolismo , Oócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Resultado do Tratamento , Ubiquinona/farmacologiaRESUMO
BACKGROUND: Adiponectin (Adipoq), a protein secreted by adipocytes in inverse proportion to the adipose mass present, modulates energy homeostasis and increases insulin sensitivity. Tissue Adipoq signaling decreases in settings of maternal diabetes, polycystic ovary syndrome (PCOS) and endometriosis, conditions which are associated with reproductive difficulty. Our objective was to define the expression and hormonal regulation of Adipoq and its receptors in the mouse preimplantation embryo and uterus. METHODS AND RESULTS: By real-time quantitative PCR, mRNA transcripts for Adipoq, AdipoR1, AdipoR2, Ppara, Ppard, FATP1 (SLC27A1) and acyl CoA oxidase (Acox1) were identified in mouse 2-cell and 8-cell embryos, while blastocyst stage embryos and trophoblast stem (TS) cells expressed mRNA for all genes except Adipoq. Protein expression of Adipoq, AdipoR1, AdipoR2, the insulin sensitive transporters GLUT8 (Slc2A8), GLUT12 (Slc2A12) and p-PRKAA1 was identified by immunofluorescence staining in all stages of preimplantation embryos including the blastocyst. In situ hybridization demonstrated the presence of Adipoq, AdipoR1 and AdipoR2 mRNA in the mouse decidual cells of the implantation site and in artificially decidualized cells, and the expression of these proteins was confirmed by western blotting. Flow cytometry confirmed cell surface expression of AdipoR1 and AdipoR2 in TS cells and decidual cells. CONCLUSIONS: These results suggest for the first time that Adipoq signaling may play an important role in preimplantation embryo development and uterine receptivity by autocrine and paracrine methods in the mouse. Implantation failures and pregnancy loss, specifically those experienced in women with maternal metabolic conditions such as diabetes, obesity and PCOS, may be the result of aberrant Adipoq and AdipoR1 and AdipoR2 expression and suboptimal decidualization in the uterus.
Assuntos
Adiponectina/metabolismo , Blastocisto/metabolismo , Decídua/metabolismo , Receptores de Adiponectina/metabolismo , Adiponectina/genética , Adiponectina/fisiologia , Animais , Western Blotting , Implantação do Embrião , Desenvolvimento Embrionário , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Hibridização In Situ , Camundongos , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Receptores de Adiponectina/genética , Receptores de Adiponectina/fisiologia , Transdução de SinaisRESUMO
Although perinatal mortality rates have improved for pregnant diabetic women because of insulin therapy and tight metabolic control, infants of diabetics still experience significantly higher rates of congenital malformations and spontaneous miscarriages compared with those of non-diabetic women. Our results here indicate that hyperglycemic conditions, either in vivo or in vitro, modulate the expression of an apoptosis regulatory gene as early as the pre-implantation blastocyst stage in the mouse. Apoptosis in the mammalian pre-implantation blastocyst is a normal process, thought to protect the early embryo by eliminating abnormal cells. Here we demonstrate that expression of Bax, a Bcl-2-like protein, is increased at the blastocyst stage in the presence of high concentrations of glucose, and that these changes correlate morphologically with increased DNA fragmentation. Expression of Bax and caspase are necessary for this in vitro glucose-induced apoptotic event, and ceramide is involved in mediating this embryotoxic effect of glucose. We also show that these apoptotic cellular changes can be prevented in vivo by treating hyperglycemic mice with insulin before and immediately after conception. These findings emphasize the importance of tight glycemic control in diabetic women at the earliest stages after conception.
Assuntos
Apoptose , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário , Hiperglicemia/complicações , Animais , Apoptose/genética , Blastocisto/metabolismo , Fragmentação do DNA , Desenvolvimento Embrionário/genética , Feminino , Camundongos , Microscopia Confocal , Microscopia de Fluorescência , Reação em Cadeia da Polimerase , Gravidez , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2RESUMO
Expression profiling of stem cells is challenging due to their small numbers and heterogeneity. The PCR colony (polony) approach has theoretical advantages as an assay for stem cells but has not been applied to small numbers of cells. An assay has been developed that is sensitive enough to detect mRNAs from small numbers of ES cells and from fractions of a single mouse blastocyst. Genes assayed include Oct3, Rex1, Nanog, Cdx2 and GLUT-1. The assay is highly sensitive so that multiple mRNAs from a single blastocyst were easily detected in the same assay. In its present version, the assay is an attractive alternative to conventional RT-PCR for profiling small populations of stem cells. The assay is also amenable to improvements that will increase its sensitivity and ability to analyze many cDNAs simultaneously.
Assuntos
Blastocisto/metabolismo , Células-Tronco Embrionárias/metabolismo , Perfilação da Expressão Gênica/métodos , RNA Mensageiro/análise , Animais , Linhagem Celular , Camundongos , Proteínas de Transporte de Cátions Orgânicos/genética , Reação em Cadeia da PolimeraseRESUMO
Congenital malformations are the leading cause of perinatal death among infants of diabetic women. Abnormal fuel metabolism and hyperglycemia have been shown to disturb embryogenesis during the earliest pre- and postimplantation stages in mice. This review presents a new model to explain, in part, adverse pregnancy outcomes associated with diabetes. In this model, by altering gene expression in developing tissues, raised glucose concentrations led to premature programmed cell death in key progenitor cells of the mouse blastocyst or in emerging organ structures in the mouse postimplantation embryo, resulting in abnormal morphogenesis or miscarriage. Although recent studies are still somewhat speculative and have currently only been explored in the mouse, this paradigm is supported by examples in other cell systems, which include human-derived cell lines, thereby suggesting that these findings are also applicable to human pregnancy.
Assuntos
Aborto Espontâneo/etiologia , Apoptose , Anormalidades Congênitas/etiologia , Hiperglicemia/complicações , Gravidez em Diabéticas/complicações , Animais , Implantação do Embrião , Desenvolvimento Embrionário , Feminino , Humanos , GravidezRESUMO
Cigarette smoke exposure causes germ cell death during spermatogenesis. Our earlier studies demonstrated that cigarette smoke condensate (CSC) causes spermatocyte cell death in vivo and growth arrest of the mouse spermatocyte cell line (GC-2spd(ts)) in vitro via the aryl hydrocarbon receptor (AHR). We hypothesize here that inactivation of AHR could prevent the CSC-induced cell death in spermatocytes. We demonstrate that CSC exposure generates oxidative stress, which differentially regulates mitochondrial apoptosis in GC-2spd(ts) and wild type (WT) and AHR knockout (AHR-KO) mouse embryonic fibroblasts (MEFs). SiRNA-mediated silencing of Ahr augments the extent of CSC-mediated cellular damage while complementing the AHR-knockout condition. Pharmacological inhibition using the AHR-antagonist (CH223191) modulates the CSC-altered expression of apoptotic proteins and significantly abrogates DNA fragmentation though the cleavage of PARP appears AHR independent. Pretreatment with CH223191 at concentrations above 50 µM significantly prevents the CSC-induced activation of caspase-3/7 and externalization of phosphatidylserine in the plasma membrane. However, MAPK inhibitors alone or together with CH223191 could not prevent the membrane damage upon CSC addition and the caspase-3/7 activation and membrane damage in AHR-deficient MEF indicates the interplay of multiple cell signaling and cytoprotective ability of AHR. Thus the data obtained on one hand signifies the protective role of AHR in maintaining normal cellular homeostasis and the other, could be a potential prophylactic therapeutic target to promote cell survival and growth under cigarette smoke exposed environment by receptor antagonism via CH223191-like mechanism. Antagonist-mediated inactivation of the aryl hydrocarbon receptor blocks downstream events leading to cigarette smoke-induced cell death of a spermatocyte cell line.
RESUMO
A majority of ovarian follicles are lost to natural death, but the disruption of factors involved in maintenance of the oocyte pool results in a further untimely follicular depletion known as premature ovarian failure. The anti-apoptotic B-cell lymphoma 2 (Bcl-2) family member myeloid cell leukemia-1 (MCL-1) has a pro-survival role in various cell types; however, its contribution to oocyte survival is unconfirmed. We present a phenotypic characterization of oocytes deficient in Mcl-1, and establish its role in maintenance of the primordial follicle (PMF) pool, growing oocyte survival and oocyte quality. Mcl-1 depletion resulted in the premature exhaustion of the ovarian reserve, characterized by early PMF loss because of activation of apoptosis. The increasingly diminished surviving cohort of growing oocytes displayed elevated markers of autophagy and mitochondrial dysfunction. Mcl-1-deficient ovulated oocytes demonstrated an increased susceptibility to cellular fragmentation with activation of the apoptotic cascade. Concomitant deletion of the pro-apoptotic Bcl-2 member Bcl-2-associated X protein (Bax) rescued the PMF phenotype and ovulated oocyte death, but did not prevent the mitochondrial dysfunction associated with Mcl-1 deficiency and could not rescue long-term breeding performance. We thus recognize MCL-1 as the essential survival factor required for conservation of the postnatal PMF pool, growing follicle survival and effective oocyte mitochondrial function.
Assuntos
Proteína de Sequência 1 de Leucemia de Células Mieloides/fisiologia , Reserva Ovariana/fisiologia , Animais , Apoptose/fisiologia , Feminino , Humanos , Camundongos , Camundongos Transgênicos , Oócitos/fisiologiaRESUMO
Women with polycystic ovary syndrome have significantly higher rates of pregnancy loss, as well as elevated insulin and IGF-1 levels. In this study, preimplantation embryos exposed to high concentrations of IGF-1 or insulin undergo extensive apoptosis of the ICM nuclei. Lack of BAX expression, the caspase inhibitor, zVAD, or the ceramide synthase inhibitor, fumonisin B1, prevents this event, suggesting involvement of programmed cell death effector pathways. In other systems, the IGF-1 concentration regulates IGF-1R expression and thus high concentrations lead to down-regulation of the receptor. Here, data show a decrease in IGF-1 receptor protein expression, both by confocal immunofluorescent microscopy and by Western analysis upon exposure to 130 nM IGF-1. Insulin-stimulated glucose uptake, an event regulated via the IGF-1 receptor, is decreased upon exposure to excess IGF-1, suggesting decreased function of the receptor. The data also show that, by blocking receptor signal transduction or by decreasing receptor expression, the apoptotic event can be recreated, thus strongly suggesting that the mechanism of high IGF-1 induced apoptosis is decreased downstream IGF-1 receptor signaling. This embryotoxic insult by high IGF-1 levels may be responsible for the high incidence of pregnancy loss seen in women with polycystic ovary syndrome.
Assuntos
Apoptose/efeitos dos fármacos , Blastocisto/citologia , Fator de Crescimento Insulin-Like I/farmacologia , Insulina/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2 , Receptor IGF Tipo 1/fisiologia , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Caspases/metabolismo , Células Cultivadas , Ceramidas/biossíntese , Tubas Uterinas/química , Feminino , Glucose/metabolismo , Marcação In Situ das Extremidades Cortadas , Insulina/análise , Fator de Crescimento Insulin-Like I/análise , Masculino , Camundongos , Camundongos Knockout , Oligonucleotídeos Antissenso/farmacologia , Proteínas Proto-Oncogênicas/deficiência , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/fisiologia , Receptor IGF Tipo 1/análise , Receptor IGF Tipo 1/antagonistas & inibidores , Proteína X Associada a bcl-2RESUMO
The diabetic state, as well as elevated culture media glucose level (950 mg D-glucose/dL) per se, significantly retards in vitro development of mouse pre-implantation embryos from a two-cell stage to blastocyst stage; maternal insulin therapy to diabetic mice reverses this impairment. This study was undertaken to assess (1) whether less extreme elevation of the media glucose concentration would also impair development, and (2) whether elevated culture media insulin or glucagon levels would alter development. Two-cell pre-embryos were recovered from B6C3F1 mice that had been stimulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hGG), mated, and killed 48 hours later. Pre-embryos were observed in culture at 24-hour intervals for a total of 72 hours at four glucose levels: 110 (n = 108), 220 (n = 101), 440 (n = 65), and 950 (n = 106) mg D-glucose/dL. Impairment in progression of development was noted at each time period; compared with development in 110 mg glucose/dL, the distribution of development was significantly different at 24 hours (chi 2 = 60.1, P less than .001), at 48 hours (chi 2 = 36.7, P less than .001), and at 72 hours (chi 2 = 45.1, P less than .001). Rate of development as assessed by ANOVA was also significantly reduced at increasing glucose levels (P less than .0001), with Duncan Multiple Range test demonstrating differences between development at higher glucose levels in the comparison of development in 110 mg/dL versus 440 mg/dL and 950 mg/dL, and at 220 mg/dL versus 950 mg/dL.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Blastocisto/fisiologia , Glucagon/farmacologia , Glucose/farmacologia , Insulina/farmacologia , Análise de Variância , Animais , Blastocisto/citologia , Blastocisto/efeitos dos fármacos , Gonadotropina Coriônica/farmacologia , Relação Dose-Resposta a Droga , Feminino , Gonadotropinas Equinas/farmacologia , Cinética , Camundongos , Camundongos Endogâmicos , Técnicas de Cultura de Órgãos , Superovulação , Fatores de TempoRESUMO
Animal models of diabetes mellitus during pregnancy have repeatedly suggested that maternal hyperglycemia was teratogenic during organogenesis, and thus may contribute to diabetic teratogenesis. However, little attention has been focused on the effects of hyperglycemia on pre-organogenic development. In this report, we examine the effect of hyperglycemia (950 mg glucose/dL) on the development of mouse pre-embryos in vitro. B6C3F1 mice were superovulated with 5 U pregnant mare serum gonadotropin (PMSG) followed by 5 U human chorionic gonadotropin (hCG) 48 hours later. Two cell pre-embryos were recovered 48 hours later, pooled together, and randomly assigned to different treatment groups. Cultures were performed in HAM's F-10 media (Gibco, Long Island, NY) with 0.1% bovine serum albumin (BSA; Sigma, St. Louis, MO) BSA at 37 degrees C in an atmosphere of 5% CO2, 5% O2, and 90% N2 with 15 to 30 embryos per milliliter of culture fluid. Cultures were viewed daily at 24, 48, and 72 hours after culturing, with recording of the development. Compared with control pre-embryos (n = 216), embryos cultured in elevated glucose levels (950 mg/dL) (n = 226) demonstrated marked growth retardation as assessed both by (1) distribution of developmental stages at each observation point (24 hours, P less than .001; 48 hours, P less than .006; 72 hours, P less than .001); and (2) a difference in the average rank sums indicating a delay in maturation (P less than .005). In a second protocol group, pre-embryos were cultured in an equivalent amount of L-glucose; no impairment in development compared with controls was noted.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Embrião de Mamíferos/embriologia , Hiperglicemia/embriologia , Gravidez em Diabéticas/embriologia , Animais , Ácido Araquidônico , Ácidos Araquidônicos/farmacologia , Blastocisto/citologia , Blastômeros/citologia , Células Cultivadas/efeitos dos fármacos , Desenvolvimento Embrionário e Fetal , Feminino , Glucose/farmacologia , Hiperglicemia/complicações , Camundongos , Óvulo/citologia , Gravidez , Gravidez em Diabéticas/metabolismo , Fatores de TempoRESUMO
OBJECTIVE: To compare pregnancy outcome after IUI versus cervical cap insemination in a donor insemination program. DESIGN: A randomized prospective clinical trial in which patients were alternately inseminated with cryopreserved human semen using either IUI or cervical cap insemination methods. SETTING: The donor insemination program at Washington University School of Medicine. PATIENTS: Forty-two women with either isolated male factor or male factor plus corrected ovulatory dysfunction using clomiphene citrate underwent 141 cycles of donor insemination. MAIN OUTCOME MEASURES: Clinical pregnancy rates (PRs) defined as a viable intrauterine gestation > 12 weeks or delivered were compared between groups using the chi 2 test. RESULTS: Clinical PRs were significantly higher in the IUI group (16.4%) compared with the cervical cap insemination group (5.9%). The spontaneous abortion rates were similar between the IUI (1.4%) and cervical cap insemination groups (4.4%). CONCLUSIONS: These findings suggest an advantage to IUI over cervical cap insemination in a donor insemination program.
Assuntos
Colo do Útero , Inseminação Artificial Heteróloga/métodos , Resultado da Gravidez , Útero , Aborto Espontâneo , Clomifeno/uso terapêutico , Criopreservação , Feminino , Humanos , Infertilidade/terapia , Masculino , Gravidez , Estudos Prospectivos , Preservação do SêmenRESUMO
This article addresses a new initiative of the Health Care Financing Administration (HCFA) to contract on a group basis with: employer self-insurance plans, unions, group health insurance companies, and Taft-Hartley Health and Welfare Funds for groups of Medicare beneficiaries. under this new concept, Medicare beneficiaries may elect in the future to obtain Medicare coverage through their group insurance plan rather than through traditional Medicare, with HCFA paying the premiums. This Medicare demonstration will bridge the gap between employer plans, which coordinate with traditional Medicare coverage, and employer-sponsored health benefit plans. This will make available to Medicare-eligible retirees similar, if not the same, managed-care alternatives as are currently available to active employees.
Assuntos
Capitação , Honorários e Preços , Planos de Assistência de Saúde para Empregados , Sistemas Pré-Pagos de Saúde/estatística & dados numéricos , Seguro Saúde , Medicare/tendências , Idoso , Centers for Medicare and Medicaid Services, U.S. , Humanos , Aposentadoria , Estados UnidosRESUMO
This chapter describes a finely tuned series of events that results in ovarian morphological changes including follicular growth, dominant follicle selection, oocyte development, ovulation, and corpus luteum formation. These changes are under the obligate control of the pituitary gonadotropins FSH and LH. The signaling mechanisms whereby the gonadotropins stimulate these changes in the ovary are now well described. The gonadotropin membrane receptors have been cloned and the cellular events proceeding from receptor binding to phosphorylation of protein gene regulators, such as CREB, have been elucidated. A whole series of intraovarian paracrine and autocrine regulators have been described as having the capability of "fine tuning" the effects of gonadotropins (see Tables I and II), and they are likely involved in dominant follicle selection and the demise of the corpus luteum. The roles of the individual paracrine regulators are, as of yet, not well known, but the IGF system offers an attractive paradigm for these intraovarian factors. Lastly, we have described the explosion of data on intracellular and intranuclear regulators. The challenge for us will be to understand the physiological significance of the seemingly unlimited number of regulatory possibilities. Much has been learned concerning the regulation of the morphological changes seen in the ovary, but much more remains to be elucidated.
Assuntos
Glândulas Endócrinas/fisiologia , Atresia Folicular/fisiologia , Hormônios/fisiologia , Folículo Ovariano/fisiologia , Ovulação/fisiologia , Óvulo/crescimento & desenvolvimento , Animais , Feminino , Humanos , Folículo Ovariano/crescimento & desenvolvimentoRESUMO
OBJECTIVE: To determine whether elevated levels of cancer antigen 125 (CA-125) correlate with the severity and clinical outcome of pelvic inflammatory disease. STUDY DESIGN: CA-125 levels were measured prospectively in 36 women with pelvic inflammatory disease using a commercial immunoassay. Initial inclusion criteria were abdominal tenderness, cervical motion tenderness and adnexal tenderness. Patients then were categorized into three groups based on the severity of the illness. Group 1 included patients with (1) temperature > or = 38.0 but < 38.5 degrees C or (2) WBC count > or = 10 but < 15 x 10(3) cells/microL. Group 2 included patients with (1) temperature > or = 38.5 degrees C or (2) WBC count > or = 15 x 10(3) cells/microL. Group 3 included patients who fulfilled the same criteria as group 2 but who also had an adnexal mass. Differences in CA-125 levels were compared statistically using the unpaired Student t test. RESULTS: All patients with the diagnosis of pelvic inflammatory disease had CA-125 levels > 30 U/mL, with a range of 48-656. Moreover, the levels were significantly different among the three groups (group 1 vs. 2, P = .002; 1 vs. 3, P = .001; 2 vs. 3, P = .001), and a correlation between severity of disease and CA-125 levels was found. All patients with a presumed pelvic abscess had CA-125 levels in the range for neoplastic processes. CONCLUSION: Elevated CA-125 levels correlate with the severity of pelvic inflammatory disease and outcome. Pelvic inflammatory disease must be considered in the differential diagnosis of elevated CA-125 levels and pelvic masses.
Assuntos
Antígeno Ca-125/sangue , Doença Inflamatória Pélvica/sangue , Adolescente , Adulto , Diagnóstico Diferencial , Feminino , Humanos , Técnicas Imunoenzimáticas , Contagem de Leucócitos , Doença Inflamatória Pélvica/diagnóstico , Doença Inflamatória Pélvica/fisiopatologia , Estudos Prospectivos , Índice de Gravidade de Doença , TemperaturaAssuntos
Células Germinativas/citologia , Receptores de Hidrocarboneto Arílico/metabolismo , Apoptose/efeitos dos fármacos , Células Germinativas/efeitos dos fármacos , Células Germinativas/metabolismo , Humanos , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Receptores de Hidrocarboneto Arílico/antagonistas & inibidores , Receptores de Hidrocarboneto Arílico/genética , Transdução de Sinais/efeitos dos fármacos , FumarRESUMO
AIMS: To evaluate the efficacy and safety of laparoscopic adrenalectomy in benign adrenal disorders. METHODS AND MATERIAL: Since July 2007, twenty patients have undergone laparoscopic adrenalectomy for various benign adrenal disorders at our institution. Every patient underwent contrast enhanced CT-abdomen. Serum corticosteroid levels were conducted in all, and urinary metanephrines, normetanephrines and VMA levels were performed in suspected pheochromocytoma. All the patients underwent laparoscopic adrenalectomy via the transperitoneal approach. RESULTS: The patients were in the age range of 18-57 years, eleven males and nine females, seven right, eleven left, two bilateral. The mean operative time was 150 minutes (120-180), mean hospital stay four days (3-5), mean intraoperative blood loss 150 ml and mean post-operative analgesic need was for 36 (24-72) hours. One out of twenty-two laparoscopic operations had to be converted into open adrenalectomy due to intra-operative complications. CONCLUSIONS: Laparoscopic adrenalectomy is a safe, effective and useful procedure without any major post-operative complication and is the gold standard for all benign adrenal disorders.
RESUMO
Glucose transport across the chondrocyte membrane is essential for chondrogenesis and the development of the skeletal system. We have previously used RT-PCR to show that fully developed human articular chondrocytes express transcripts for the GLUT1 and GLUT9 glucose transporters. In this study we report on the expression and immunohistochemical localization of the GLUT1 and GLUT9 proteins in embryonic and mature ovine cartilage. We also provide Western blot evidence for GLUT1 and GLUT9 expression in mature ovine chondrocytes. Ovine embryos (developmental stages E32 to E36 and E42 to E45) were obtained from pregnant ewes humanely killed by injection with sodium pentobarbitone. Embryos were fixed and processed for immunohistochemistry. Polyclonal antibodies to GLUT1 and GLUT9 revealed that both transporters are expressed in developing chondrocytes in ovine embryos and in the superficial, middle and deep layers of ovine cartilage from mature animals. GLUT1 expression was observed in erythrocytes and organs including heart, liver, and kidney. GLUT9 was also found in heart, kidney and liver. Western blotting confirmed the presence of the GLUT1 protein which migrated between the 50 and 64 kDa markers and two specific GLUT9 bands migrating under the 50 and 60 kDa markers, respectively. The presence of GLUT1 and GLUT9 in developing joints of ovine embryos suggests that these proteins may be important in glucose delivery to developing chondroblasts. Expression of these GLUT isoforms may be an important bioenergetic adaptation for chondrocytes in the extracellular matrix of developing cartilage.
Assuntos
Condrócitos/metabolismo , Proteínas de Transporte de Monossacarídeos/biossíntese , Animais , Western Blotting , Cartilagem Articular/citologia , Cartilagem Articular/embriologia , Cartilagem Articular/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Transportador de Glucose Tipo 1 , Técnicas Imunoenzimáticas , Ovinos/embriologiaRESUMO
From animal and human studies is it clear that mammalian embryos are vulnerable to injury during the earliest preimplantation stages of development. Exposure to some agents during this brief period has resulted not only in fetal loss but also in malformations. Thus, the potential for maternal induced embryotoxicity exists earlier than previously expected. This period is marked by a drastic change in metabolic needs at the late morula stage. Glucose becomes the predominant exogenous energy substrate and enters the blastocyst via one of three facilitative glucose transporters, GLUT1, GLUT2, and GLUT3. It has been shown that maternal diabetes adversely affects the preimplantation embryo. Recent work has revealed that hyperglycemia leads to a downregulation of the GLUTs at the blastocyst stage in the mouse. This downregulation results in decreased glucose transport into the blastocyst of diabetic mice and thus lower intraembryonic free glucose levels. The embryos are starving themselves of the key substrate necessary for survival. Maternal diabetes also causes a decrease in the number of cells in rat blastocyst and recently hyperglycemia has been shown to induce apoptosis in the mouse blastocyst via cell death effector pathways involving BAX and caspases. Significant loss of key progenitor cells from the blastocyst may predispose these diabetic embryos to later developmental deficiencies manifesting as dysmorphogenesis, fetal loss or early growth delay. The hypothesis that the hyperglycemia-induced decrease in glucose transport triggers apoptosis is presented and discussed as a novel mechanism to explain preimplantation diabetic embryopathy.