Association of in vitro fertilization outcome with circulating insulin-like growth factor components prior to cycle initiation
American journal of obstetrics and gynecology, Jan 30, 2015
Components of the insulin-like growth factor (IGF) system enhance in vitro embryo quality and imp... more Components of the insulin-like growth factor (IGF) system enhance in vitro embryo quality and implantation rates in both animal models and human in vitro fertilization (IVF). We evaluated whether differences in serum levels of these components in women prior to initiation of an IVF cycle would be predictive of subsequent outcome. In this retrospective study sera from women obtained at day 2 of their IVF cycle (at baseline before stimulation) were assayed for IGF-I, IGF-II, and IGF binding protein (BP)-1 by enzyme-linked immunosorbent assay. Samples from 54 women with a live birth, 38 with a transient biochemical pregnancy, 45 with a spontaneous abortion, 54 who did not become pregnant, and 35 who had an ectopic pregnancy were available for analysis. Associations between the assays and outcome were evaluated by the Kruskal-Wallis test and receiver operating characteristic analysis. There were no differences in the number of oocytes retrieved, oocyte quality, fertilization rates, or e...
Detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction in introital specimens from pregnant women
American Journal of Obstetrics and Gynecology, 1996
OBJECTIVE: We analyzed whether specimens obtained from the vaginal introitus were comparable to e... more OBJECTIVE: We analyzed whether specimens obtained from the vaginal introitus were comparable to endocervical and posterior vaginal vault specimens for detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction. STUDY DESIGN: Introital and endocervical specimens were obtained from 300 women at the first prenatal visit. Specimens from the posterior vaginal vault were also obtained from 219 of these
Comparative distribution of murine mammary tumor virus (mumtv) anti- body, antigen (gp52) and antigen-antibody complexes in sera of mam- mary tumor bearing and non-tumor bearing c3h/bi mice. Abstr
Autologous Endometrial Coculture in Patients with In Vitro-Fertilization (IVF) Failure: Correlations of Outcome with Leukemia Inhibiting Factor (LIF) Production1
American Journal of Reproductive Immunology, 2001
To analyze the effectiveness of autologous endometrial coculture (AECC) in improving embryo quali... more To analyze the effectiveness of autologous endometrial coculture (AECC) in improving embryo quality and pregnancy rates in 1,030 consecutive cycles of in vitro fertilization-embryo transfer (IVF-ET) utilizing AECC from January 1996 to December 2001. Embryos from each of 1,030 patients allocated to growth on AECC were analyzed for outcome. All patients had previously undergone failed IVF cycles. During a luteal phase biopsy (5-12 days after the luteinizing hormone surge) performed prior to the treatment cycle, glandular (G) and stromal (S) endometrial cells were isolated by enzymatic digestion and separated based on differential sedimentation rates. These cells were cryopreserved, then plated as a 50%/50% combination of G and S cells prior to embryo exposure. The conditioned medium was changed every 2 days. Embryos were randomly grown on endometrial coculture (ECC) or conventional media if > 6 oocytes were normally fertilized. Otherwise, all embryos were grown on AECC. The patients' mean age was 36.9 (+/-3.1) years. The patients had on average a history of 3.1 (+/- 1.7) failed prior attempts. When comparing a previous cycle (same institution only), the cleaved embryos on day 3 were of an improved quality (6.8+/-1.2 vs. 5.5+/-1.0 blastomeres and 14.6% +/- 9.3 vs. 27.2% +/- 9.8 fragmentation, P <.05). Twenty-two (2.13%) patients did not undergo ET secondary to poor embryonic development. Overall positive and clinical pregnancy rates of 49.8% and 41.5% were noted, respectively. Age remained the most important predictor of outcome. We demonstrated a significant improvement in embryo quality with ECC. We also demonstrated that patients with a poor prognosis secondary to prior IVF failures can have a good outcome when utilizing AECC.
International journal of cancer. Journal international du cancer, 2015
Inhibition of autophagy is a characteristic of ovarian cancer. We determined whether inhibition o... more Inhibition of autophagy is a characteristic of ovarian cancer. We determined whether inhibition of autophagy by vaginal fluid could provide a non-invasive test for cancer risk stratification in women presenting with an adnexal mass. Vaginal fluid supernatants from 90 women undergoing evaluation for a suspicious adnexal mass were incubated with peripheral blood mononuclear cells (PBMCs) obtained from healthy women under conditions that induce autophagy. Rapamycin, an autophagy inducer, was added to some cultures. After 48 hours the cells were collected, lysed and assayed by ELISA for intracellular p62 concentration. p62 is a cytoplasmic protein that is consumed during autophagy induction. Its concentration is inversely proportional to the extent of autophagy induction. Clinical information including pathological diagnoses was obtained after completion of laboratory studies. Mean p62 levels were 9.4 ng/ml in the 21 women with a subsequent malignant diagnosis, 4.5 ng/ml in the 8 women ...
Background: For more than 100 years, group A Streptococcus has been identified as a cause of seve... more Background: For more than 100 years, group A Streptococcus has been identified as a cause of severe and, in many cases, fatal infections of the female urogenital tract. Due to advances in hospital hygiene and the advent of antibiotics, this type of infection has been virtually eradicated. However, within the last three decades there has been an increase in severe intraand post-partum infections attributed to GAS.
Background: Streptococcus agalactiae (group B Streptococcus) is a bacterial pathogen that causes ... more Background: Streptococcus agalactiae (group B Streptococcus) is a bacterial pathogen that causes severe intrauterine infections leading to fetal morbidity and mortality. The pathogenesis of GBS infection in this environment is poorly understood, in part because we lack a detailed understanding of the adaptation of this pathogen to growth in amniotic fluid. To address this knowledge deficit, we characterized the transcriptome of GBS grown in human amniotic fluid (AF) and compared it with the transcriptome in rich laboratory medium.
The objective of this work was to evaluate the influence of human papillomavirus (HPV) vaccinatio... more The objective of this work was to evaluate the influence of human papillomavirus (HPV) vaccination on peripheral blood mononuclear cell (PBMC) proliferation and cytokine gene transcription. PBMCs isolated after HPV immunization were incubated with HPV vaccine, phytohemagglutinin, or buffer. Cell proliferation was assessed by MTT reduction assay. RNA was extracted from PBMCs, and the relative concentration of cytokine messenger RNA (mRNA) transcripts (IFN-β, IFN-γ, IL-12, TNF-α, IL-6, IL-17, or IL-10) relative to transcription of the β-actin gene was determined by real-time polymerase chain reaction. PBMC proliferation in response to HPV vaccine and PHA were greater than that observed in unstimulated cells (p<0.001). Cytokine mRNAs were upregulated in stimulated PBMC cultures. The median increase in vaccine-stimulated cultures was: IFN-β=334.4-fold; IL-12=46.33-fold; IFN-γ=12.64-fold; IL-6=9.07-fold; IL-17=7.33-fold; IL-10=6.47-fold; and TNF-α=2.36-fold. The IFN-β expression was s...
Interleukin 1 receptor antagonist gene polymorphism in women with vulvar vestibulitis
American journal of obstetrics and gynecology, 2000
Vulvar vestibulitis is a chronic inflammatory syndrome of unknown cause and pathogenesis. We exam... more Vulvar vestibulitis is a chronic inflammatory syndrome of unknown cause and pathogenesis. We examined the relation between vulvar vestibulitis and polymorphisms in the gene coding for the interleukin 1 receptor antagonist, a naturally occurring down-regulator of proinflammatory immune responses. Cells from the lower genital tract of 68 women with vulvar vestibulitis, 343 women with no history of vulvodynia, and 40 women with human papillomavirus cervical infection were tested by polymerase chain reaction for the different alleles of the gene encoding for interleukin 1 receptor antagonist. The presence of human papillomavirus in the specimens was determined by polymerase chain reaction with the use of degenerate consensus primers to the conserved L1 gene. Allele 2 of the gene encoding the interleukin 1 receptor antagonist was present in homozygous form in 52.9% of women with vulvar vestibulitis. In marked contrast only 8. 5% of the control women and 2.5% of women with human papilloma...
Objective: Women with diminished ovarian reserve have high rates of pregnancy loss. A number of t... more Objective: Women with diminished ovarian reserve have high rates of pregnancy loss. A number of tests have been described that attempt to determine ovarian reserve. Serum inhibin B concentration has been used to predict orarian reserve as well as pregnancy outcome after in virto fertilization (IVF) cycle outcomes. The present study was undertaken to assess inhibin B serum concentrations among women experiencing recurrent pregnancy loss.
Pregnancy Hypertension: An International Journal of Women's Cardiovascular Health, 2015
Preeclampsia (PE) is a specific disorder of pregnancy associated with hyperuricemia, higher level... more Preeclampsia (PE) is a specific disorder of pregnancy associated with hyperuricemia, higher levels of pro-inflammatory cytokines, enhanced leukocyte activation and oxidative stress. Adenosine deaminase (ADA) is an enzyme present in cells of the immune system, and is considered an indicator of cellular inflammation. This study aimed to compare serum levels of ADA with TNF-alpha and IL-1beta produced by peripheral blood mononuclear cells (PBMC) in pregnant women with PE, in normotensive pregnant (NT) and in non-pregnant (NP) women. Sixty women with PE and 30 NT paired by gestational age were evaluated. Twenty healthy NP women matched for age with pregnant women were included as a control group. PBMC were obtained from the three groups studied and cultured with or without lipopolysaccharide (LPS) for 18h at 37°C, and TNF-alpha and IL-1beta production was assessed in the supernatant of cultures by enzyme-linked immunosorbent assay. ADA and uric acid serum concentrations were determined by colorimetric methods. Differences between groups were analyzed by non-parametric tests with a significance level of 5%. Serum levels of ADA were significantly higher in PE group compared with NT and NP groups. A positive correlation between ADA and uric acid levels was detected in preeclamptic women (r=0.4082; p=0.0024). There was no significant difference in relation to ADA levels when PE patients were classified in early and late-onset PE. The endogenous production of IL-1beta and TNF-alpha by PBMC was significantly higher in PE group than in NT and NP women, showing the activation state of these cells in PE. The increased levels of ADA associated with hyperuricemia and proinflammatory cytokines suggest that this enzyme plays an important role in the systemic inflammation observed in PE. V.S. Giorgi: None. V.T. Borges: None. S.S. Witkin: None. C.F. Bannwart-Castro: None. J.C. Peracoli: None. M.T. Peracoli: None.
Pregnancy Hypertension: An International Journal of Women's Cardiovascular Health, 2015
Preeclampsia (PE) is a clinical complication of pregnancy associated with high levels of inflamma... more Preeclampsia (PE) is a clinical complication of pregnancy associated with high levels of inflammatory cytokines, leukocyte activation and oxidative stress. Autophagy, an intracellular catabolic process, which removes damaged organelles and cytoplasmic proteins seems to be able to maintain cellular homeostasis. Moreover, it also has a potent anti-inflammatory mechanism by destroying inflammasomes, an intracellular multi-protein structure required for processing and releasing bioactive Interleukin-1 beta (IL-1beta). The present study compared the ability of sera from pregnant women with or without preeclampsia to induce autophagy in peripheral blood mononuclear cells (PBMC), and the relationship between this activity and serum concentrations of uric acid, IL-1beta, IL-10 and TNF-alpha. PBMC from 20 healthy non-pregnant women were incubated with sera from 45 pregnant women, being 23 normotensive (NT) and 22 with PE in the presence or absence of rapamycin, an autophagy inducer. After 48h, the cells were assayed for p62, a cytoplasmic protein associated with autophagy inhibition. Sera from both groups of pregnant women were tested for IL-1beta, IL-10 and TNF-alpha by enzyme-linked immunosorbent assay. Autophagy was induced to a greater extent by sera from women with PE than by NT pregnant women. The relationship between inflammatory cytokines in sera of preeclamptic pregnant women showed association between autophagy induction (decrease in cytoplasmic p62) and higher concentrations of IL-1beta (r= -0.4949; p=0.0192), and TNF-alpha (r=- 0.5302; p=0.0111). This relationship was absent in sera from NT women. There was no association between IL-10 or uric acid levels and autophagy both in preeclamptic and NT group. Association between autophagy induction and higher levels of inflammatory cytokines in sera from patients with preeclampsia suggests that the enhanced pro-inflammatory state characteristic of preeclampsia may result in the up-regulation of autophagy induction in these women. Financial support: Fapesp 2014/00693-9. M.L. Matias: None. M.T. Peracoli: None. T.T. Kanninen: None. G. Sisti: None. M. Romao: None. I.C. Weel: None. J.C. Peracoli: None. A.M. Bongiovanni: None. S.S. Witkin: None.
Infectious Diseases in Obstetrics & Gynecology, 1996
The 60kD heat shock protein (hsp60) is a highly conserved protein and a dominant antigen of most ... more The 60kD heat shock protein (hsp60) is a highly conserved protein and a dominant antigen of most pathogenic bacteria. In some women, chronic or repeated upper genital tract infections with Chlamydia trachomatis, and possibly with other microorganisms, induces immune sensitization to epitopes of hsp60 that are present in both the microbial and human hsp60. Once a woman becomes sensitized to these conserved epitopes, any subsequent induction of human or bacterial hsp60 expression will reactivate hsp60-sensitized lymphocytes and initiate a pro-inflammatory immune response. Hsp60 is expressed during the early stages of pregnancy, by both the embryo and the maternal decidua. We examined, therefore, whether women who were sensitized to hsp60 experi- enced less successful pregnancy outcomes compared to women who were not sensitized to this antigen. In women undergoing in vitro fertilization (IVF), the presence of cervical IgA antibodies reactive with the C. trachomatis hsp60 correlated with implantation failure after embryo transfer. Further analysis revealed that an immunodominant epitope for these IgA antibodies was an hsp60 epitope shared between C. trachomatis and man. In subsequent studies of women not undergoing IVF, cervical IgA antibodies to the human hsp60 were identified in 13 of 91 reproductive age women.
Vulvodynia is a prevalent and highly distressing disorder, with major consequences for interperso... more Vulvodynia is a prevalent and highly distressing disorder, with major consequences for interpersonal and psychologic well-being. Due to the high level of psychologic distress commonly associated with the disorder, as well as the absence of evidencebased guidelines for clinical practice, a multinational consensus conference on the topic of vulvodynia was convened. The first National Institutes of Health (NIH)sponsored conference on this topic was held in 1997, shortly after which a program of funded vulvodynia research was initiated. A second NIH meeting was convened in 2003 to present an overview of the science and epidemiology of vulvodynia with a focus on the fundamental mechanisms of this pain disorder.
To evaluate the gene expression of Toll-Like (TLR-2 and TLR-4) receptors and cytokine profile in ... more To evaluate the gene expression of Toll-Like (TLR-2 and TLR-4) receptors and cytokine profile in postmenopausal women with or without metabolic syndrome (MetS). In this cross-sectional study, 311 Brazilian women (age≥45 years and amenorrhea≥12 months) were included. Women showing three or more of the following diagnostic criteria were diagnosed as positive for MetS: waist circumference&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;88 cm, triglycerides≥150 mg/dL, HDL cholesterol&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;50 mg/dL, blood pressure≥130/85 mmHg, and fasting glucose≥100 mg/dL. The expression of TLR-2 and TLR-4 in peripheral blood was evaluated by RNA extraction and subsequent real time PCR analysis. The cytokine profile, tumor necrosis factor alpha (TNF-α) and interleukins 1β, 6, and 10, were measured by ELISA. The expression of TLR-2 RNA was demonstrated in 32.5% and TLR-4 in 20.6% of the subjects. There was no association between the expression of TLR-2 and TLR-4 and the presence or absence of MetS (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). A greater production of IL-6 was associated with TLR-2 and TLR-4 expressions and greater production of TNF-α was associated only with TLR-2 expression (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). Only the lower quartile of IL-10 was associated with the presence of the MetS (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). TLR-2 and TLR-4 expressions were associated with increased pro-inflammatory cytokines, IL-6 and TNF-α, with no association with biomarkers of MetS. The low concentrations of IL-10 may suggest an anti-inflammatory modulation in postmenopausal women with MetS.
Objective: To evaluate the efficacy of a self-collected vaginal introital specimen, obtained by w... more Objective: To evaluate the efficacy of a self-collected vaginal introital specimen, obtained by women using only an instruction booklet as a guide, for the detection of Chlamydia tvuchomatis using polymerase chain reaction (PCR).
Association of in vitro fertilization outcome with circulating insulin-like growth factor components prior to cycle initiation
American journal of obstetrics and gynecology, Jan 30, 2015
Components of the insulin-like growth factor (IGF) system enhance in vitro embryo quality and imp... more Components of the insulin-like growth factor (IGF) system enhance in vitro embryo quality and implantation rates in both animal models and human in vitro fertilization (IVF). We evaluated whether differences in serum levels of these components in women prior to initiation of an IVF cycle would be predictive of subsequent outcome. In this retrospective study sera from women obtained at day 2 of their IVF cycle (at baseline before stimulation) were assayed for IGF-I, IGF-II, and IGF binding protein (BP)-1 by enzyme-linked immunosorbent assay. Samples from 54 women with a live birth, 38 with a transient biochemical pregnancy, 45 with a spontaneous abortion, 54 who did not become pregnant, and 35 who had an ectopic pregnancy were available for analysis. Associations between the assays and outcome were evaluated by the Kruskal-Wallis test and receiver operating characteristic analysis. There were no differences in the number of oocytes retrieved, oocyte quality, fertilization rates, or e...
Detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction in introital specimens from pregnant women
American Journal of Obstetrics and Gynecology, 1996
OBJECTIVE: We analyzed whether specimens obtained from the vaginal introitus were comparable to e... more OBJECTIVE: We analyzed whether specimens obtained from the vaginal introitus were comparable to endocervical and posterior vaginal vault specimens for detection of Chlamydia trachomatis and Trichomonas vaginalis by polymerase chain reaction. STUDY DESIGN: Introital and endocervical specimens were obtained from 300 women at the first prenatal visit. Specimens from the posterior vaginal vault were also obtained from 219 of these
Comparative distribution of murine mammary tumor virus (mumtv) anti- body, antigen (gp52) and antigen-antibody complexes in sera of mam- mary tumor bearing and non-tumor bearing c3h/bi mice. Abstr
Autologous Endometrial Coculture in Patients with In Vitro-Fertilization (IVF) Failure: Correlations of Outcome with Leukemia Inhibiting Factor (LIF) Production1
American Journal of Reproductive Immunology, 2001
To analyze the effectiveness of autologous endometrial coculture (AECC) in improving embryo quali... more To analyze the effectiveness of autologous endometrial coculture (AECC) in improving embryo quality and pregnancy rates in 1,030 consecutive cycles of in vitro fertilization-embryo transfer (IVF-ET) utilizing AECC from January 1996 to December 2001. Embryos from each of 1,030 patients allocated to growth on AECC were analyzed for outcome. All patients had previously undergone failed IVF cycles. During a luteal phase biopsy (5-12 days after the luteinizing hormone surge) performed prior to the treatment cycle, glandular (G) and stromal (S) endometrial cells were isolated by enzymatic digestion and separated based on differential sedimentation rates. These cells were cryopreserved, then plated as a 50%/50% combination of G and S cells prior to embryo exposure. The conditioned medium was changed every 2 days. Embryos were randomly grown on endometrial coculture (ECC) or conventional media if &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; 6 oocytes were normally fertilized. Otherwise, all embryos were grown on AECC. The patients&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; mean age was 36.9 (+/-3.1) years. The patients had on average a history of 3.1 (+/- 1.7) failed prior attempts. When comparing a previous cycle (same institution only), the cleaved embryos on day 3 were of an improved quality (6.8+/-1.2 vs. 5.5+/-1.0 blastomeres and 14.6% +/- 9.3 vs. 27.2% +/- 9.8 fragmentation, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;.05). Twenty-two (2.13%) patients did not undergo ET secondary to poor embryonic development. Overall positive and clinical pregnancy rates of 49.8% and 41.5% were noted, respectively. Age remained the most important predictor of outcome. We demonstrated a significant improvement in embryo quality with ECC. We also demonstrated that patients with a poor prognosis secondary to prior IVF failures can have a good outcome when utilizing AECC.
International journal of cancer. Journal international du cancer, 2015
Inhibition of autophagy is a characteristic of ovarian cancer. We determined whether inhibition o... more Inhibition of autophagy is a characteristic of ovarian cancer. We determined whether inhibition of autophagy by vaginal fluid could provide a non-invasive test for cancer risk stratification in women presenting with an adnexal mass. Vaginal fluid supernatants from 90 women undergoing evaluation for a suspicious adnexal mass were incubated with peripheral blood mononuclear cells (PBMCs) obtained from healthy women under conditions that induce autophagy. Rapamycin, an autophagy inducer, was added to some cultures. After 48 hours the cells were collected, lysed and assayed by ELISA for intracellular p62 concentration. p62 is a cytoplasmic protein that is consumed during autophagy induction. Its concentration is inversely proportional to the extent of autophagy induction. Clinical information including pathological diagnoses was obtained after completion of laboratory studies. Mean p62 levels were 9.4 ng/ml in the 21 women with a subsequent malignant diagnosis, 4.5 ng/ml in the 8 women ...
Background: For more than 100 years, group A Streptococcus has been identified as a cause of seve... more Background: For more than 100 years, group A Streptococcus has been identified as a cause of severe and, in many cases, fatal infections of the female urogenital tract. Due to advances in hospital hygiene and the advent of antibiotics, this type of infection has been virtually eradicated. However, within the last three decades there has been an increase in severe intraand post-partum infections attributed to GAS.
Background: Streptococcus agalactiae (group B Streptococcus) is a bacterial pathogen that causes ... more Background: Streptococcus agalactiae (group B Streptococcus) is a bacterial pathogen that causes severe intrauterine infections leading to fetal morbidity and mortality. The pathogenesis of GBS infection in this environment is poorly understood, in part because we lack a detailed understanding of the adaptation of this pathogen to growth in amniotic fluid. To address this knowledge deficit, we characterized the transcriptome of GBS grown in human amniotic fluid (AF) and compared it with the transcriptome in rich laboratory medium.
The objective of this work was to evaluate the influence of human papillomavirus (HPV) vaccinatio... more The objective of this work was to evaluate the influence of human papillomavirus (HPV) vaccination on peripheral blood mononuclear cell (PBMC) proliferation and cytokine gene transcription. PBMCs isolated after HPV immunization were incubated with HPV vaccine, phytohemagglutinin, or buffer. Cell proliferation was assessed by MTT reduction assay. RNA was extracted from PBMCs, and the relative concentration of cytokine messenger RNA (mRNA) transcripts (IFN-β, IFN-γ, IL-12, TNF-α, IL-6, IL-17, or IL-10) relative to transcription of the β-actin gene was determined by real-time polymerase chain reaction. PBMC proliferation in response to HPV vaccine and PHA were greater than that observed in unstimulated cells (p<0.001). Cytokine mRNAs were upregulated in stimulated PBMC cultures. The median increase in vaccine-stimulated cultures was: IFN-β=334.4-fold; IL-12=46.33-fold; IFN-γ=12.64-fold; IL-6=9.07-fold; IL-17=7.33-fold; IL-10=6.47-fold; and TNF-α=2.36-fold. The IFN-β expression was s...
Interleukin 1 receptor antagonist gene polymorphism in women with vulvar vestibulitis
American journal of obstetrics and gynecology, 2000
Vulvar vestibulitis is a chronic inflammatory syndrome of unknown cause and pathogenesis. We exam... more Vulvar vestibulitis is a chronic inflammatory syndrome of unknown cause and pathogenesis. We examined the relation between vulvar vestibulitis and polymorphisms in the gene coding for the interleukin 1 receptor antagonist, a naturally occurring down-regulator of proinflammatory immune responses. Cells from the lower genital tract of 68 women with vulvar vestibulitis, 343 women with no history of vulvodynia, and 40 women with human papillomavirus cervical infection were tested by polymerase chain reaction for the different alleles of the gene encoding for interleukin 1 receptor antagonist. The presence of human papillomavirus in the specimens was determined by polymerase chain reaction with the use of degenerate consensus primers to the conserved L1 gene. Allele 2 of the gene encoding the interleukin 1 receptor antagonist was present in homozygous form in 52.9% of women with vulvar vestibulitis. In marked contrast only 8. 5% of the control women and 2.5% of women with human papilloma...
Objective: Women with diminished ovarian reserve have high rates of pregnancy loss. A number of t... more Objective: Women with diminished ovarian reserve have high rates of pregnancy loss. A number of tests have been described that attempt to determine ovarian reserve. Serum inhibin B concentration has been used to predict orarian reserve as well as pregnancy outcome after in virto fertilization (IVF) cycle outcomes. The present study was undertaken to assess inhibin B serum concentrations among women experiencing recurrent pregnancy loss.
Pregnancy Hypertension: An International Journal of Women's Cardiovascular Health, 2015
Preeclampsia (PE) is a specific disorder of pregnancy associated with hyperuricemia, higher level... more Preeclampsia (PE) is a specific disorder of pregnancy associated with hyperuricemia, higher levels of pro-inflammatory cytokines, enhanced leukocyte activation and oxidative stress. Adenosine deaminase (ADA) is an enzyme present in cells of the immune system, and is considered an indicator of cellular inflammation. This study aimed to compare serum levels of ADA with TNF-alpha and IL-1beta produced by peripheral blood mononuclear cells (PBMC) in pregnant women with PE, in normotensive pregnant (NT) and in non-pregnant (NP) women. Sixty women with PE and 30 NT paired by gestational age were evaluated. Twenty healthy NP women matched for age with pregnant women were included as a control group. PBMC were obtained from the three groups studied and cultured with or without lipopolysaccharide (LPS) for 18h at 37°C, and TNF-alpha and IL-1beta production was assessed in the supernatant of cultures by enzyme-linked immunosorbent assay. ADA and uric acid serum concentrations were determined by colorimetric methods. Differences between groups were analyzed by non-parametric tests with a significance level of 5%. Serum levels of ADA were significantly higher in PE group compared with NT and NP groups. A positive correlation between ADA and uric acid levels was detected in preeclamptic women (r=0.4082; p=0.0024). There was no significant difference in relation to ADA levels when PE patients were classified in early and late-onset PE. The endogenous production of IL-1beta and TNF-alpha by PBMC was significantly higher in PE group than in NT and NP women, showing the activation state of these cells in PE. The increased levels of ADA associated with hyperuricemia and proinflammatory cytokines suggest that this enzyme plays an important role in the systemic inflammation observed in PE. V.S. Giorgi: None. V.T. Borges: None. S.S. Witkin: None. C.F. Bannwart-Castro: None. J.C. Peracoli: None. M.T. Peracoli: None.
Pregnancy Hypertension: An International Journal of Women's Cardiovascular Health, 2015
Preeclampsia (PE) is a clinical complication of pregnancy associated with high levels of inflamma... more Preeclampsia (PE) is a clinical complication of pregnancy associated with high levels of inflammatory cytokines, leukocyte activation and oxidative stress. Autophagy, an intracellular catabolic process, which removes damaged organelles and cytoplasmic proteins seems to be able to maintain cellular homeostasis. Moreover, it also has a potent anti-inflammatory mechanism by destroying inflammasomes, an intracellular multi-protein structure required for processing and releasing bioactive Interleukin-1 beta (IL-1beta). The present study compared the ability of sera from pregnant women with or without preeclampsia to induce autophagy in peripheral blood mononuclear cells (PBMC), and the relationship between this activity and serum concentrations of uric acid, IL-1beta, IL-10 and TNF-alpha. PBMC from 20 healthy non-pregnant women were incubated with sera from 45 pregnant women, being 23 normotensive (NT) and 22 with PE in the presence or absence of rapamycin, an autophagy inducer. After 48h, the cells were assayed for p62, a cytoplasmic protein associated with autophagy inhibition. Sera from both groups of pregnant women were tested for IL-1beta, IL-10 and TNF-alpha by enzyme-linked immunosorbent assay. Autophagy was induced to a greater extent by sera from women with PE than by NT pregnant women. The relationship between inflammatory cytokines in sera of preeclamptic pregnant women showed association between autophagy induction (decrease in cytoplasmic p62) and higher concentrations of IL-1beta (r= -0.4949; p=0.0192), and TNF-alpha (r=- 0.5302; p=0.0111). This relationship was absent in sera from NT women. There was no association between IL-10 or uric acid levels and autophagy both in preeclamptic and NT group. Association between autophagy induction and higher levels of inflammatory cytokines in sera from patients with preeclampsia suggests that the enhanced pro-inflammatory state characteristic of preeclampsia may result in the up-regulation of autophagy induction in these women. Financial support: Fapesp 2014/00693-9. M.L. Matias: None. M.T. Peracoli: None. T.T. Kanninen: None. G. Sisti: None. M. Romao: None. I.C. Weel: None. J.C. Peracoli: None. A.M. Bongiovanni: None. S.S. Witkin: None.
Infectious Diseases in Obstetrics & Gynecology, 1996
The 60kD heat shock protein (hsp60) is a highly conserved protein and a dominant antigen of most ... more The 60kD heat shock protein (hsp60) is a highly conserved protein and a dominant antigen of most pathogenic bacteria. In some women, chronic or repeated upper genital tract infections with Chlamydia trachomatis, and possibly with other microorganisms, induces immune sensitization to epitopes of hsp60 that are present in both the microbial and human hsp60. Once a woman becomes sensitized to these conserved epitopes, any subsequent induction of human or bacterial hsp60 expression will reactivate hsp60-sensitized lymphocytes and initiate a pro-inflammatory immune response. Hsp60 is expressed during the early stages of pregnancy, by both the embryo and the maternal decidua. We examined, therefore, whether women who were sensitized to hsp60 experi- enced less successful pregnancy outcomes compared to women who were not sensitized to this antigen. In women undergoing in vitro fertilization (IVF), the presence of cervical IgA antibodies reactive with the C. trachomatis hsp60 correlated with implantation failure after embryo transfer. Further analysis revealed that an immunodominant epitope for these IgA antibodies was an hsp60 epitope shared between C. trachomatis and man. In subsequent studies of women not undergoing IVF, cervical IgA antibodies to the human hsp60 were identified in 13 of 91 reproductive age women.
Vulvodynia is a prevalent and highly distressing disorder, with major consequences for interperso... more Vulvodynia is a prevalent and highly distressing disorder, with major consequences for interpersonal and psychologic well-being. Due to the high level of psychologic distress commonly associated with the disorder, as well as the absence of evidencebased guidelines for clinical practice, a multinational consensus conference on the topic of vulvodynia was convened. The first National Institutes of Health (NIH)sponsored conference on this topic was held in 1997, shortly after which a program of funded vulvodynia research was initiated. A second NIH meeting was convened in 2003 to present an overview of the science and epidemiology of vulvodynia with a focus on the fundamental mechanisms of this pain disorder.
To evaluate the gene expression of Toll-Like (TLR-2 and TLR-4) receptors and cytokine profile in ... more To evaluate the gene expression of Toll-Like (TLR-2 and TLR-4) receptors and cytokine profile in postmenopausal women with or without metabolic syndrome (MetS). In this cross-sectional study, 311 Brazilian women (age≥45 years and amenorrhea≥12 months) were included. Women showing three or more of the following diagnostic criteria were diagnosed as positive for MetS: waist circumference&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;88 cm, triglycerides≥150 mg/dL, HDL cholesterol&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;50 mg/dL, blood pressure≥130/85 mmHg, and fasting glucose≥100 mg/dL. The expression of TLR-2 and TLR-4 in peripheral blood was evaluated by RNA extraction and subsequent real time PCR analysis. The cytokine profile, tumor necrosis factor alpha (TNF-α) and interleukins 1β, 6, and 10, were measured by ELISA. The expression of TLR-2 RNA was demonstrated in 32.5% and TLR-4 in 20.6% of the subjects. There was no association between the expression of TLR-2 and TLR-4 and the presence or absence of MetS (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). A greater production of IL-6 was associated with TLR-2 and TLR-4 expressions and greater production of TNF-α was associated only with TLR-2 expression (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). Only the lower quartile of IL-10 was associated with the presence of the MetS (P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.05). TLR-2 and TLR-4 expressions were associated with increased pro-inflammatory cytokines, IL-6 and TNF-α, with no association with biomarkers of MetS. The low concentrations of IL-10 may suggest an anti-inflammatory modulation in postmenopausal women with MetS.
Objective: To evaluate the efficacy of a self-collected vaginal introital specimen, obtained by w... more Objective: To evaluate the efficacy of a self-collected vaginal introital specimen, obtained by women using only an instruction booklet as a guide, for the detection of Chlamydia tvuchomatis using polymerase chain reaction (PCR).
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