RESUMO
BACKGROUND: Posttraumatic stress disorder (PTSD) is declared in 3 to 6 % of postpartum women (PP) and up to 18.5 % in cases of complications of pregnancy or childbirth. The objective of this study is to assess the prevalence of PTSD after a red code cesarean section and to identify the risk factors among the prenatal vulnerability factors, the birth alert factors and the maintenance factors in PP. METHOD: A phone or computerized questionnaire including an Questionnaire de stress immédiat and the Posttraumatic Stress Disorder Checklist for DSM-5 was offered to patients who had a red code cesarean section between 05/12/2015 and 02/28/2021 at the University South Hospital of Reunion Island. RESULTS: Among the 555 cesarean sections selected, 329 parturients responded. The prevalence of PTSD was 20.1 % and was stable over time. The 2 risk factors found were the negative experience of childbirth and the proven traumatic experience. Prenatal vunerability factors were not found to be statistically significant. Almost 3 in 4 women had not been informed of the risk of cesarean section and more than 1 in 2 women did not have an explanation in PP. CONCLUSION: Red code cesarean sections cause PTSD in 1 in 5 women. This lasting disorder can last up to 6 years after childbirth. This indicates the seriousness of this disorder and the need to prevent it. The risk of developing it is 4 times greater in the event of a traumatic experience proven in the Questionnaire de stress immédiat. Offering this questionnaire in the maternity could be an important element of secondary prevention. The role of health personnel remains essential.
Assuntos
Transtornos de Estresse Pós-Traumáticos , Cesárea/efeitos adversos , Feminino , Humanos , Incidência , Parto , Gravidez , Fatores de Risco , Transtornos de Estresse Pós-Traumáticos/epidemiologia , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Syphilis is a sexually transmitted infection (STI) with a global prevalence estimated at 0.5% in 2012. Syphilis has been on the rise among men who have sex with men (MSM) in high-income countries and remains at endemic levels in low- and middle-income countries. This trend, however, has not been observed in Reunion Island. OBJECTIVES: To determine the prevalence, clinical characteristics and risk factors of syphilis in at-risk patients visiting the South Reunion STI clinic in Reunion Island. METHODS: This monocentric cross-sectional study included all patients who visited our STI clinic between 2017 and 2020. Syphilis serology was performed on all included patients, and data were collected using a standardized self-administered questionnaire. RESULTS: Over the 3-year study period, 2593 patients were enrolled. The prevalence of syphilis was 7.52% (n = 195, 95% CI, 6.50-8.65%) in the overall study population, 11.76% (n = 18, 95% CI, 6.97-18.59%) in minors (aged under 18 years) and 36.36% (n = 16, 95% CI, 21-59%) in pregnant women. The risk factors identified in multivariate analysis were being female [adjusted Prevalence Ratio (aPR) 1.85, 95% CI, 1.10-3.11], being MSM (aPR 2.87, 95% CI, 1.71-4.80), being aged under 18 years (aPR 3.54, 95% CI, 1.90-6.57), living in precarious conditions [aPR 3.12, 95% CI, 2.11-4.62] and being born in Reunion Island (aPR 2.43, 95% CI, 1.42-4.13). The clinical presentation was heterogeneous (plaques and papules, chancre, atypical ulcerations, multiple ulcerations, condyloma lata, etc.). CONCLUSIONS: These findings suggest a high prevalence of syphilis in at-risk patients visiting our STI clinic. Unlike the situation in other high-income countries, the people most at risk of syphilis in Reunion Island are local-born residents, minors, women and precarious patients. This is a source of concern, especially given the risk of resurgence of congenital syphilis on the island.
Assuntos
Infecções por HIV , Minorias Sexuais e de Gênero , Infecções Sexualmente Transmissíveis , Sífilis , Adolescente , Idoso , Estudos Transversais , Feminino , Homossexualidade Masculina , Humanos , Masculino , Menores de Idade , Gravidez , Prevalência , Reunião/epidemiologia , Fatores de Risco , Infecções Sexualmente Transmissíveis/epidemiologia , Sífilis/epidemiologiaRESUMO
OBJECTIVE: We aim to evaluate the knowledge and physicians' practices concerning fertility preservation in women with endometriosis. DESIGN: Descriptive, observational, national study using an online self-questionnaire, sent by email to French gynaecologists in October 2019 within 2 months. RESULTS: We obtained 110 analyzable responses from mainly surgeons (54 %) and reproductive clinicians (19 %) with a good experience (average 15 years of practice). Amongst these practitioners, 91 % seemed aware of latest French recommendations on endometriosis issued in December 2017. The most commonly used surgical techniques for management of endometriomas were intra-peritoneal cystectomy (51 %), vaporization by plasma energy (29 %) and destruction by bipolar coagulation (8.5 %). Preoperative AMH was systematically or often prescribed by 78 % of the practitioners against 37.3 % who did it postoperatively. Furthermore, 74 % also considered and performed fertility preservation strategy to manage endometriosis. It was offered in situations of bilateral or recurrent endometrioma, but only 33 % offered it in unilateral endometrioma cases. In the cases recorded, vitrification of mature oocytes appears to be the most common fertility preservation technique (used by 87 % of the practitioners). CONCLUSION: We observed in our population of sensitized practitioners a good and adequate knowledge concerning endometriosis physiopathology and recommendations for its management, with good information delivery to women. Operating techniques are adapted although information and education concerning fertility preservation indications seem necessary. The place of multidisciplinary concertation meeting in endometriosis appears essential both for discussion of surgical indications and for fertility preservation possibilities. Creation of dedicated structures should be encouraged.
Assuntos
Endometriose/terapia , Preservação da Fertilidade , Padrões de Prática Médica/estatística & dados numéricos , Adulto , Idoso , Competência Clínica , Feminino , França , Humanos , Masculino , Pessoa de Meia-Idade , Inquéritos e QuestionáriosRESUMO
In vitro fertilization (IVF) is nowadays a reliable and common method for couple who need medically assisted procreation. Complications are rare. We report in this paper, the case of a woman with severe endometriosis who developed ureteral obstruction complicated by a renal rupture of the fornix due to ovarian hyperstimulation during an IVF attempt. The condition was diagnosed by CT scan and resolved with insertion of double-J catheter in the left ureter.
Assuntos
Fertilização in vitro/efeitos adversos , Nefropatias/etiologia , Síndrome de Hiperestimulação Ovariana/complicações , Ruptura Espontânea/etiologia , Obstrução Ureteral/etiologia , Adulto , Feminino , Humanos , Nefropatias/diagnóstico por imagem , Nefropatias/terapia , Ruptura Espontânea/diagnóstico por imagem , Ruptura Espontânea/terapia , Obstrução Ureteral/diagnóstico por imagem , Obstrução Ureteral/terapiaRESUMO
We report a rare case of primary gallbladder adenocarcinoma producing human chorionic gonadotropin (HCG) in a 31-year-old woman. The patient was first misdiagnosed and monitored for an extra-uterine pregnancy. The most frequent cause of elevated serum HCG is pregnancy but elevated HCG can also be a marker of others pathologies like tumors. It is of utmost importance to keep in mind all the possible causes of elevated serum HCG. Once pregnancy has been ruled out, complementary exams should be performed to seek a tumor, especially since tumors producing HCG can be particularly aggressive.
Assuntos
Carcinoma/sangue , Gonadotropina Coriônica/sangue , Neoplasias da Vesícula Biliar/sangue , Adulto , Feminino , HumanosRESUMO
OBJECTIVE: Inhibiting bacterial biofilms is of major significance for proper wound healing. The choice of the dressing material plays a key role, as bacteria can live in dressings and keep reinfecting the wound. This study examines the effectiveness of a colloidal silver gel (Ag-gel) wound dressing in inhibiting the growth of bacteria in a mouse wound model. METHOD: Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii and two different meticillin-resistant Staphylococcus aureus (MRSA) strains were examined. Bacteria were measured in vitro on the dressing, and in vivo studies were carried out to analyses both the dressing and the infected tissue. RESULTS: Using colony-forming unit (CFU) assays, over 7 logs of inhibition (100%) were found for Staphylococcus aureus, Pseudomonas aeruginosa and Acinetobacter baumannii for the Ag-gel dressing when compared with the control dressing. In vivo, complete inhibition was observered for the three most common bacteria on the Ag-gel dressing and the tissue under that dressing. These results were confirmed by an in vivo live imaging system. However, with MRSA strains, only 2-3 logs of inhibition were recorded. CONCLUSION: The Ag-gel was effective in preventing biofilm infections caused by both Gram-negative and Gram-positive bacteria.
Assuntos
Acinetobacter baumannii/efeitos dos fármacos , Anti-Infecciosos Locais/farmacologia , Bandagens , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/efeitos dos fármacos , Prata/farmacologia , Infecções Estafilocócicas/tratamento farmacológico , Infecção dos Ferimentos/tratamento farmacológico , Animais , Anti-Infecciosos Locais/uso terapêutico , Biofilmes/efeitos dos fármacos , Modelos Animais de Doenças , Géis , Técnicas In Vitro , Camundongos , Prata/uso terapêutico , Staphylococcus aureus/efeitos dos fármacosRESUMO
UNLABELLED: Cervical incompetency is one of the direct causes of neonatal morbidity and mortality; a unique and efficient treatment of which is cervical cerclage. The objective of this study was the evaluation of physicians' practice patterns concerning cerclage in Reunion Island, in order to reinforce the management and information of patients at risk. The indications and complications of cerclages effectuated in 2010 and 2011 were compared to the literature. MATERIAL AND METHODS: In this retrospective study, all the medical records of cerclage realized in Reunion Island during two years were collected and analyzed, specifically data concerning patients' cerclage, the complications, and the outcome of the pregnancy. RESULTS: We listed 200 cerclages, which were predominantly prophylactic cerclages (75.5%) and represented 0.71% of all births. A total of 71% of the indications of cerclage in Reunion Island did not take into account the recommendations of the literature. Analysis revealed the frequent use of prophylactic cerclage and subsequently reflected the insufficient use of therapeutic cerclage. In those cases, the rate of premature delivery was indeed lower (P=0.003), as well as the rate of chorioamniotitis (P=0.003). CONCLUSION: Cerclage is an efficient treatment to extend the length of the pregnancy. Nevertheless, it is important to comply with the recommendations given by the literature, by spotting the patients at risk of premature delivery, and recommend cerclage only in case of real cervical incompetency, for the sake of improving their management and reducing the rate of complications.
Assuntos
Aborto Espontâneo/epidemiologia , Cerclagem Cervical/estatística & dados numéricos , Guias de Prática Clínica como Assunto , Padrões de Prática Médica/estatística & dados numéricos , Nascimento Prematuro/epidemiologia , Incompetência do Colo do Útero/epidemiologia , Adolescente , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Gravidez , Estudos Retrospectivos , Reunião/epidemiologia , Incompetência do Colo do Útero/cirurgia , Adulto JovemRESUMO
OBJECTIVES: Partial mastectomy, augmentation and reduction mammaplasty are often operated on women who are not yet bothered by breastfeeding. The objectives of this study were to evaluate the information given to patients before surgery, and describe difficulties that mothers confront when starting breastfeeding in order to create a reference document about breastfeeding to inform patients who will undergo such surgery in the future. MATERIAL AND METHODS: We led one first study to evaluate the surgeons' practice in the Reunion Island and a second retrospective and descriptive study upon patients. RESULTS: We encountered the fact that few patients in childbearing age ask for information about breastfeeding before undergoing surgery, but surgeons do not systematically give such information either, even less before partial mastectomy. The impact of surgery on breastfeeding depends on the type of intervention and the surgical technique. Even though breastfeeding is possible, the mean period of breastfeeding after surgery is shorter and the most frequent difficulty encountered is lactation insufficiency, even more after reduction mammaplasty, periareolar incision, and nipple hypoesthesia after surgery. DISCUSSION AND CONCLUSION: The information document that we tried to establish concerning breastfeeding after partial mastectomy, augmentation and reduction mammaplasty, may compensate patients' lack of information and sums up all the complications described in our study and in the literature.
Assuntos
Aleitamento Materno , Mama/cirurgia , Feminino , Humanos , Consentimento Livre e Esclarecido , Mamoplastia/efeitos adversos , Mastectomia Segmentar , Mamilos/cirurgia , Educação de Pacientes como Assunto , Gravidez , Estudos Retrospectivos , ReuniãoRESUMO
Few studies address chronic disease risk for Southeast Asians in the United States. In 1999, the authors conducted a cross-sectional study of bone mineral density (BMD) estimated from ultrasonic calcaneal measurements in women born in Southeast Asia who then lived in Chicago, Illinois. The study addressed three questions: Do Southeast-Asian women have relatively low BMD? What factors before and after immigration are associated with BMD? Are factors that reflect the childhood/adolescent environment equally associated with BMD for postmenopausal and premenopausal women? An interviewer-administered bilingual questionnaire collected immigration, reproductive, and lifestyle data from 213 women (aged 20--80 years) born in Vietnam, Cambodia, or Laos. The authors found that the estimated mean BMD of postmenopausal Southeast-Asian women was lower than the reference values for White women. Four summary indicators of childhood/adolescent environment were predictive of higher BMD: more years of education, earlier age at menarche, lower height, and coastal birth; these indicators were more strongly associated with BMD for premenopausal (multiple-partial R(2) = 0.21) than postmenopausal (R(2) = 0.06) women. Young-adult exposures (e.g., early first pregnancy and age at immigration) and proximal lifestyle factors (e.g., smoking, physical inactivity, vegetarian diet, and betel nut use) were also assessed as potential predictors of BMD.
Assuntos
Densidade Óssea , Calcâneo/diagnóstico por imagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Antropometria , Sudeste Asiático/etnologia , Chicago/epidemiologia , Estudos Transversais , Feminino , Humanos , Análise dos Mínimos Quadrados , Estilo de Vida , Pessoa de Meia-Idade , Fatores de Risco , Inquéritos e Questionários , UltrassonografiaRESUMO
Gene therapy for hepatocellular carcinoma (HCC) has shown some promise, but its evaluation requires relevant experimental models. With this aim, we present an evaluation of the interest of using the woodchuck model of HCC to assess in vivo gene transfer efficiency. We tested the transduction efficacy of the adenoviral vectors directing lacZ gene product expression under the control of the cytomegalovirus and alpha-fetoprotein (AFP) regulatory sequences. We have also investigated whether an adenoviral cytomegalovirus-thymidine kinase (Tk) vector might induce an antitumoral effect in this model. Our results demonstrate that with direct intratumoral and intrahepatic arterial injections, transduction of a significant proportion of tumor cells occurred even in large HCC nodules. Furthermore, due to intra-arterial anastomoses, direct intratumoral injection led to transduction of some noninjected HCC nodules. Moreover, direct intratumoral injection of a herpes simplex virus-1 Tk-encoding vector induced, on ganciclovir administration, a significant antitumoral effect in the two animals evaluated. However, in one animal, massive hepatic failure occurred due to Tk expression in nontumor cells. These results emphasize the need to target the expression of the Tk gene to tumor cells using a hepatoma-specific promoter such as AFP promoter. However, we showed that, in vivo, lacZ expression as driven by the AFP promoter was extremely low, thus emphasizing some potential pitfalls when using this approach. Altogether, our data stress the need to test gene therapy-based strategies in such in vivo animal models of HCC and evaluate gene transduction, expression, and biological activity, as well as its potential toxicity.
Assuntos
Adenoviridae/genética , Carcinoma Hepatocelular/terapia , Ganciclovir/uso terapêutico , Terapia Genética/métodos , Neoplasias Hepáticas Experimentais/terapia , Simplexvirus/enzimologia , Timidina Quinase/genética , Transdução Genética , Animais , Antivirais/uso terapêutico , Antivirais/toxicidade , Apoptose , Carcinoma Hepatocelular/metabolismo , Terapia Combinada , Citomegalovirus/enzimologia , Modelos Animais de Doenças , Ganciclovir/toxicidade , Vetores Genéticos , Óperon Lac , Fígado/metabolismo , Neoplasias Hepáticas Experimentais/metabolismo , Marmota , Necrose , Regiões Promotoras Genéticas , beta-Galactosidase/biossínteseRESUMO
Adenoviral vectors are known to transduce hepatocytes in normal liver tissue with high efficiency. The aim of this study was to investigate whether sinusoidal endothelial cells, which separate hepatocytes from the bloodstream in the sinusoidal lumen, are permissive for infection by adenoviruses. We show here that microvascular liver sinusoidal endothelial cells are not infected by adenovirus type 5 in vivo or in vitro unless high MOIs are used. In contrast, macrovascular endothelial cells from aorta are efficiently infected by adenovirus type 5. In addition, Kupffer cells, similar to sinusoidal endothelial cells, are not infected by adenovirus type 5. Liver sinusoidal endothelial cells do not express the integrin receptor alpha(v)beta3, which is required for efficient infection by adenoviruses. Our results demonstrate that hepatocytes are the main cell population of the liver that is infected by adenovirus type 5.
Assuntos
Adenoviridae/crescimento & desenvolvimento , Endotélio/virologia , Células de Kupffer/virologia , Fígado/virologia , Animais , Células Cultivadas , Endotélio/anatomia & histologia , Endotélio/citologia , Humanos , Fígado/anatomia & histologia , Fígado/citologia , CamundongosRESUMO
Transfer of the herpes simplex virus-thymidine kinase (HSV-tk) gene followed by the administration of ganciclovir (GCV) into hepatocellular carcinoma (HCC)-derived cell lines either in vitro or transplanted into nude mice has been shown to provide a potential strategy for HSV-tk-based gene therapy of HCC. We report herein an analysis of the antitumoral efficacy of two recombinant adenoviruses (Ads), Ad.CMVtk and Ad.AFPtk, in a relevant model of multifocal hepatic lesions induced in rats by a potent alkylating chemical carcinogen, diethylnitrosamine. Two routes of administration of the Ad were studied: intratumoral and intrahepatic artery injections. Both recombinant Ads, Ad.CMVtk and Ad.AFPtk, express the HSV-tk gene under the control of the early enhancer/promoter cytomegalovirus and alpha-fetoprotein regulatory gene sequences, respectively. The antitumor response was assessed by magnetic resonance imaging and by autopsy and histological analysis following postmortem. Tumor growth cessation was demonstrated by magnetic resonance imaging in large tumor nodules of size 5-8 mm treated by intratumoral administration of 2x10(9) pfu Ad.CMVtk plus i.p. treatment with GCV. We also show an antitumor efficacy in small tumor nodules of size <3 mm treated with 2x10(9) pfu Ad.CMVtk plus GCV by the intrahepatic artery route, albeit associated with an adverse toxicity. In vivo targeting of the HSV-tk gene to diethylnitrosamine-induced HCC cells with the recombinant Ad.AFPtk suppresses the hepatic toxicity in the nontumoral liver. The lower antitumor response would argue for the use of multiple injections of such adenoviral constructs. These observations may lead to potential approaches for designing gene therapy destined for early treatment of dysplastic nodules or advanced HCC in cirrhosis.
Assuntos
Adenoviridae/genética , Terapia Genética , Neoplasias Hepáticas Experimentais/terapia , Simplexvirus/enzimologia , Timidina Quinase/genética , Animais , Dietilnitrosamina , Ganciclovir/efeitos adversos , Ganciclovir/uso terapêutico , Terapia Genética/efeitos adversos , Artéria Hepática , Humanos , Masculino , Ratos , Ratos Wistar , Células Tumorais CultivadasRESUMO
Adenovirus-mediated gene therapy of experimental hepatocarcinoma is hindered by low transduction efficacy in vivo. We evaluated the extent of gene expression following various routes of administration of recombinant adenovirus AdCMVlacZ in diethylnitrosamine-induced rat hepatocarcinoma. We first characterized the vascularization of diethylnitrosamine-induced hepatocarcinomas using a computerized tomography scanner approach. The efficacy of gene transfer was then evaluated by three routes of administration: intraportal, selective injection through the hepatic artery and direct injection into the tumor. Diethylnitrosamine-induced hepatocarcinomas had predominantly an arterial blood supply, 67% of the total liver blood supply. Compared with intraportal administration, arterial injection improved gene transfer into tumors whereas that to the non-tumor areas was diminished. In addition, this route of injection allowed the efficient transduction of dysplastic nodules. Diethylnitrosamine-induced hepatocarcinoma in rats is a relevant model for the study of human hepatocarcinoma due to its vascularization. Arterial infusion improved the ratio of transduced tumorous to nontumorous cells and allowed targeting of gene transfer to dysplastic nodules. This will be useful in the design of gene therapy for hepatocarcinoma.
Assuntos
Adenoviridae , Carcinoma Hepatocelular/terapia , Técnicas de Transferência de Genes , Vetores Genéticos , Neoplasias Hepáticas Experimentais/terapia , Animais , Carcinoma Hepatocelular/diagnóstico por imagem , Dietilnitrosamina , Expressão Gênica , Artéria Hepática , Injeções Intra-Arteriais , Neoplasias Hepáticas Experimentais/diagnóstico por imagem , Masculino , Ratos , Ratos Wistar , Tomografia Computadorizada por Raios X , beta-Galactosidase/genéticaRESUMO
A serious limitation in the use of the DNA-cleaving, antitumoral-antibiotic, bleomycin during chemotherapy is pulmonary toxicity. Lung injury induced by bleomycin is characterized by an increased deposition of interstitial extracellular matrix proteins in the alveolar wall that compromises respiratory function. Several drugs have been tested in animal models to prevent the pulmonary toxicity of bleomycin, but have not led to a useful clinical treatment because of their adverse effects on other tissues. We have shown that transgenic mice expressing Streptoalloteichus hindustanus (Sh) ble bleomycin resistance protein in pulmonary epithelial cells in the lungs are protected against bleomycin-induced toxicity in lungs. In the present study, we used intranasal administration by adenovirus-mediated gene transfer of the bleomycin resistance Sh ble gene to mouse lung for prevention of bleomycin-induced pulmonary fibrosis. We constructed recombinant adenoviruses Ad.CMVble and Ad.RSVble harboring the bleomycin resistance Sh ble gene under the control of the cytomegalovirus early promoter and the Rous sarcoma virus early promoter, respectively. Transgene expression was detected in epithelia of conducting airways and alveolar septa by immunostaining with a rabbit polyclonal antibody directed against the bleomycin resistance protein and persisted for the duration of drug treatment; i.e., up to 17 d. No toxic effect was seen in adenovirus-treated mice. Pretreatment of mice with Ad.CMVble or Ad.RSVble completely prevented collagen deposition 42-133 d after bleomycin treatment, as measured by lung OH-proline content. Histologic studies indicated that there was little or no lung injury in the adenovirus/bleomycin-treated mice compared with the bleomycin-treated mice. These observations may lead to new approaches for the prevention of bleomycin-induced pulmonary fibrosis.
Assuntos
Acetiltransferases , Adenovírus Humanos/fisiologia , Bleomicina , Técnicas de Transferência de Genes , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/prevenção & controle , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Brônquios/química , Brônquios/enzimologia , Resistência Microbiana a Medicamentos/genética , Epitélio/química , Epitélio/enzimologia , Feminino , Vetores Genéticos , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Prolina/efeitos dos fármacos , Fibrose Pulmonar/patologia , Streptomyces/genética , beta-Galactosidase/genéticaRESUMO
Thymic epithelium is involved in negative selection, but its precise role in selecting the CD4 T cell repertoire remains elusive. By using two transgenic mice, we have investigated how medullary thymic epithelium (mTE) and bone marrow (BM)-derived cells contribute to tolerance of CD4 T cells to nuclear beta-galactosidase (beta-gal). CD4 T cells were not tolerant when beta-gal was expressed in thymic BM-derived cells. In contrast, CD4 T cells of mice expressing beta-gal in mTE were tolerized. Tolerance resulted from presentation of endogenous beta-gal by mTE cells but not from cross-priming. mTE cells presented nuclear beta-gal to a Th clone in vitro, while thymic dendritic cells did not. The data indicate that mTE but not thymic BM-derived cells can use a MHC class II endogenous presentation pathway to induce tolerance to nuclear proteins.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Tolerância Imunológica , Proteínas Nucleares/imunologia , Timo/imunologia , Animais , Apresentação de Antígeno/genética , Células Epiteliais , Epitélio/imunologia , Centro Germinativo/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Tolerância Imunológica/genética , Interleucina-2/farmacologia , Ativação Linfocitária/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Camundongos Transgênicos , Proteínas Nucleares/genética , Proteínas Recombinantes/farmacologia , Timo/anatomia & histologia , beta-Galactosidase/genéticaRESUMO
The low molecular weight B cell growth factor (LMW-BCGF) induces the G1 --> S transition in human B lymphocytes activated by a first signal, Staphylococcus aureus Cowan (SAC) or anti-mu antibody. It also stimulates proliferation of normal long-term B cell lines and some B cell tumors. We have previously reported that LMW-BCGF induces the hydrolysis of polyphosphoinositides (PI) and a rise in intracellular free calcium concentration, through the generation of inositol trisphosphate (InsP3) (Renard et al., J. Immunol. 18, 1705, 1988). In the present work we have analyzed the possible association between early signaling events elicited by LMW-BCGF in SAC-activated B cells and its ability to provoke DNA synthesis, notably at the level of phospholipase C (PLC) and protein kinase C (PK-C) activation. Inhibitors of PLC and of InsP3-induced calcium release were found to block LMW-BCGF-dependent DNA synthesis. An increase in membrane-associated protein kinase C (PK-C) activity was detected after the addition of the growth factor and the mitogenic effect of LMW-BCGF was partially suppressed when B cell blasts were incubated with staurosporine or H-3, two inhibitors of PK-C activity. In addition, the mitogenic effect due to the addition of LMW-BCGF was not modified by the incubation of B cell blasts with high concentrations of TPA, even if this treatment inhibited cellular response to a low concentration of TPA. LMW-BCGF also increased intracellular pH in B cell blasts and lymphokine-induced mitogenic activity was reduced when the Na+/H+ amiloride or ethylisopropyl amiloride (EIPA) antiport blockers were added. These results suggest that (i) LMW-BCGF-induced PI breakdown and CA2+ mobilization and cell alkalinization are associated with the induction of cell proliferation, and (ii) the activation of PK-C does not appear to be the sole pathway activated by LMW-BCGF.
Assuntos
Linfócitos B/imunologia , Ativação Enzimática/efeitos dos fármacos , Linfocinas/farmacologia , Mitose/efeitos dos fármacos , Proteína Quinase C/metabolismo , Transdução de Sinais , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina , Alcaloides , Amilorida , Linfócitos B/efeitos dos fármacos , Proteínas de Transporte/antagonistas & inibidores , DNA/biossíntese , Humanos , Concentração de Íons de Hidrogênio , Isoquinolinas , Linfocinas/antagonistas & inibidores , Neomicina , Piperazinas , Trocadores de Sódio-Hidrogênio , Estaurosporina , Fosfolipases Tipo C/antagonistas & inibidoresRESUMO
The four fluorescent derivatives of TPA--dansylaza-TPA, NBDaza-TPA, and (N)- and (P)-dansylamino-TPA--were synthesized and examined for their ability to induce differentiation in human promyelocytic leukemic HL60 cells. At a concentration of 20 nM, all the derivatives inhibited proliferation and induced 60-80% of the cells to differentiate into macrophage-like cells. Removal of dansylaza-TPA from the medium after 5 h did not arrest adherence or the expression of nonspecific esterase activity. However, upon removal of any of the other three compounds after 5 h, HL60 cells became nonadherent and expressed low nonspecific esterase activity after additional culture. To investigate the relationship between protein kinase C (PKC) activation and cell maturation, PKC activity and translocation were measured after 0.5, 5, 24, and 48 h of treatment with each compound. Cells induced to differentiate by dansylaza-TPA or (N)- or (P)-dansylamino-TPA exhibited enhanced PKC activity, 50-80% of which was located in the particulate fraction. In cells that differentiated with NBDaza-TPA, 65-70% of PKC activity remained in the cytosol. After removal of the TPA derivatives, all cells exhibited PKC activity in the cytosol. These results indicate that the fluorescent derivatives are as potent as TPA in inducing HL60 cell differentiation. However, in the case of NBDaza-TPA and (N)- or (P)-dansylamino-TPA, their continuous presence in the culture medium was required for the recruitment of cells to differentiate. Consequently, it is suggested that activation and translocation of PKC are among the early biochemical events that trigger HL60 cell differentiation. Nevertheless, these two events alone are not sufficient to induce differentiation.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Proteína Quinase C/metabolismo , Acetato de Tetradecanoilforbol/análogos & derivados , Adesão Celular/efeitos dos fármacos , Humanos , Acetato de Tetradecanoilforbol/farmacologia , Células Tumorais CultivadasRESUMO
Dansyl-TPA, a fluorescent TPA analogue, which is a label with a high affinity for C3H/10T1/2 cells and induces 3H-choline release from these cells (Tran et al. Nouv. J. Chim 1984, 8, 751-757), has been studied for in vitro promoting activity in the same cell line initiated by a carcinogen MNNG and in vivo short-term mouse skin tests. In vitro, dansyl-TPA expresses transforming effect in its own (without MNNG pretreatment) as well as increases the production of transformed foci in MNNG-treated cells. In in vivo skin tests, dansyl-TPA displays lower effects than TPA on mouse skin. These results indicate a low promoting potential of dansyl-TPA.