RESUMO
OBJECTIVES: To evaluate the relationship between the attitude of the fetal head quantified by means of the chin-to-chest angle (CCA) in fetuses in occiput posterior (OP) position at the beginning of the second stage of labor, and persistent OP position at birth. METHODS: This was a single-center, prospective observational study conducted at the University Hospital of Parma, Parma, Italy. We included singleton pregnancies at term with fetuses in the OP position at the beginning of the second stage of labor. The fetal head position, station by means of angle of progression and head-to-perineum distance, and attitude by means of CCA were assessed using transabdominal or transperineal ultrasound. The primary outcome was persistent OP position at birth. RESULTS: Between January and July 2022, 76 women were included in the study. There were 48 (63.2%) spontaneous rotations of the fetal head and spontaneous vaginal delivery occurred in all. Among the 28 (36.8%) fetuses that did not rotate spontaneously into an occiput anterior position, eight (28.6%) had a spontaneous vaginal delivery, while operative vaginal delivery and Cesarean delivery was performed in 11 (39.3%) and nine (32.1%) cases, respectively. Multivariable logistic regression analysis showed that the CCA (adjusted odds ratio (aOR), 2.15 (95% CI, 1.22-3.78); P = 0.008) and nulliparity (aOR, 0.20 (95% CI, 0.06-0.76); P = 0.02) were associated independently with persistent OP position at birth. Moreover, the CCA showed an area under the receiver-operating-characteristics curve of 0.69 (95% CI, 0.56-0.82); P = 0.005) for the prediction of persistent OP position. The optimal cut-off value of the CCA was 36.5°, and was associated with a sensitivity of 0.82 (95% CI, 0.63-0.94), specificity of 0.50 (95% CI, 0.35-0.65), positive predictive value of 0.49 (95% CI, 0.34-0.64), negative predictive value of 0.83 (95% CI, 0.64-0.94), positive likelihood ratio of 1.64 (95% CI, 1.18-2.29) and negative likelihood ratio of 0.36 (95% CI, 0.15-0.83). CONCLUSIONS: Our data show that, within a population of women with fetal OP position at the beginning of the second stage of labor, the sonographic fetal head attitude measured by means of the CCA might help in the identification of fetuses at risk of persistent OP position. Such findings can be useful for patient counseling when OP position is diagnosed at full cervical dilatation. Further studies should investigate if the CCA might select patients who may benefit from manual rotation of the fetal head. © 2023 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.
Assuntos
Feto , Apresentação no Trabalho de Parto , Recém-Nascido , Gravidez , Feminino , Humanos , Estudos Prospectivos , Feto/diagnóstico por imagem , Segunda Fase do Trabalho de Parto , Ultrassonografia Pré-Natal , Parto Obstétrico , Cabeça/diagnóstico por imagemRESUMO
OBJECTIVES: Restorative properties of medicinal plants such as Genista sessilifolia DC. have often been suggested to occur, in epidemiological studies. However, full characterization of effective principles responsible for this action has never previously been performed. Here, we have characterized G. sessilifolia's anti-cancer effects and identified the chemical components involved in this anti-tumour action. MATERIALS AND METHODS: Cell cycle, apoptosis, necrosis, differentiation analyses, high-performance liquid chromatography, western blotting, RNA extraction, real-time PCR and primers have all been observed/used in the study. RESULTS: We report that G. sessilifolia methanol extract has anti-cancer activity on solid and haematological cancer cells. G. sessilifolia extract's anti-proliferative action is closely bound to induction of apoptosis, whereas differentiation is only weakly modulated. Analysis of G. sessilifolia extract, by high-performance liquid chromatography, identifies fraction 18-22 as the pertinent component for induction of apoptosis, whereas fractions 11-13 and 27-30 both seem to contribute to differentiation. G. sessilifolia extract induces apoptosis mediated by caspase activation and p21, Rb, p53, Bcl2-associated agonist of cell death (BAD), tumour necrosis factor receptor super-family, member 10 (TRAIL) overexpression and death receptor 5 (DR5). Accordingly, fraction 18-22 inducing apoptosis was able to induce TRAIL. CONCLUSIONS: Our results indicate that G. sessilifolia extract and its fraction 18-22 containing genistin and isoprunetin, were able to induce anti-cancer effects supporting the hypothesis of a pro-apoptotic intrinsic content of this natural medicinal plant.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose , Genista/química , Extratos Vegetais/farmacologia , Antineoplásicos Fitogênicos/química , Caspase 8/química , Caspase 8/genética , Ciclo Celular , Diferenciação Celular , Proliferação de Células/efeitos dos fármacos , Fracionamento Químico/métodos , Cromatografia Líquida de Alta Pressão , Ativação Enzimática , Citometria de Fluxo , Genisteína/química , Genisteína/isolamento & purificação , Genisteína/farmacologia , Granulócitos/efeitos dos fármacos , Granulócitos/patologia , Células HeLa , Humanos , Isoflavonas/química , Isoflavonas/isolamento & purificação , Isoflavonas/farmacologia , Células MCF-7 , Metanol/química , Componentes Aéreos da Planta/química , Extratos Vegetais/química , Reação em Cadeia da Polimerase em Tempo Real , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/química , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/genética , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Células U937 , Proteína de Morte Celular Associada a bcl/genética , Proteína de Morte Celular Associada a bcl/metabolismoRESUMO
OBJECTIVES: Curative properties of medicinal plants such as Psidium guajava L. (Myrtaceae) have often been indicated by epidemiological studies on populations in which these fruits are consumed daily. However, complete characterization of the active principles responsible for this ability has never been performed. Here, we have characterized P. guajava's anti-cancer potential and identified the parts of the fruit involved in its anti-neoplastic action. MATERIALS AND METHODS: We studied morphology of our cells, cell cycle characteristics and apoptosis and performed immunostaining, differentiation and western blot analyses. RESULTS: We report that the P. guajava extract exerted anti-cancer control on both haematological and solid neoplasias. P. guajava extract's anti-tumour properties were found to be tightly bound to induction of apoptosis and differentiation. Use of ex vivo myeloid leukaemia blasts corroborated that P. guajava was able to induce cell death but did not exhibit anti-cancer effects on all malignant cells investigated, indicating selective activity against certain types of tumour. Analyses of P. guajava pulp, peel and seeds identified the pulp as being the most relevant component for causing cell cycle arrest and apoptosis, whereas peel was responsible for causing cell differentiation. P. guajava itself and its pulp-derived extract were found to induce apoptosis accompanied by caspase activation and p16, p21, Fas ligand (FASL TNF super-family, member 6), Bcl-2-associated agonist of cell death (BAD) and tumour necrosis factor receptor super-family, member 10b (DR5), overexpression. CONCLUSIONS: Our findings showed that P. guajava L. extract was able to exert anti-cancer activity on cultures in vitro and ex vivo, supporting the hypothesis of its anti malignant pro-apoptotic modulation.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Fitoterapia , Psidium , Caspases/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Leucemia Promielocítica Aguda/tratamento farmacológico , Leucemia Promielocítica Aguda/metabolismo , Leucemia Promielocítica Aguda/patologia , Proteínas de Neoplasias/metabolismo , Extratos Vegetais/farmacologia , Plantas Medicinais , Células U937RESUMO
Recently, aqueous solutions polluted by BPA have been bioremediated by us using laccase immobilized on hydrophobic membranes in non-isothermal bioreactors. BPA degradation was checked using analytical methods. To assess in vitro the occurred bioremediation, the proliferation and viability indexes of MCF-7 cells incubated in the presence of aqueous solutions of BPA, or of enzyme-treated BPA solutions, have been measured as a function of the initial BPA concentration. The results demonstrated that: i) at each initial BPA concentration used, both the proliferation and viability indexes are a function of the duration of enzyme treatment; ii) proliferation and viability are uncoupled biological processes with respect to BPA enzyme treatment. Non-isothermal bioreactors are a useful tool for the bioremediation of aqueous solutions polluted by BPA, which is an example of an endocrine disruptor that belongs to the alkyl phenol family.
Assuntos
Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Disruptores Endócrinos/metabolismo , Disruptores Endócrinos/toxicidade , Lacase/metabolismo , Fenóis/metabolismo , Fenóis/toxicidade , Compostos Benzidrílicos , Linhagem Celular Tumoral , Humanos , Fenóis/antagonistas & inibidoresRESUMO
BACKGROUND: The retinoblastoma-interacting zinc-finger gene RIZ is expressed in two forms (RIZ1 and RIZ2) that differ for the presence near the N-terminus of RIZ1 of a conserved domain, defined PR (PRDI-BF1-RIZ homology), homologous to a similar domain present in other proteins recognized as tumor suppressor gene products. The RIZ1 form is usually absent or expressed at low levels in tumor cells, whereas RIZ2 is frequently expressed. We investigated a possible involvement of RIZ1 in differentiation control using a myeloid cell maturation model that is easily modulated by retinoids and other agents. MATERIALS AND METHODS: HL60 or NB4 cell lines or patients' leukemic promyelocytes were treated with all- trans -retinoic acid or other agents to induce differentiation. RIZ gene expression was determined with reverse transcriptase polymerase chain reaction (RT-PCR) and RNase protection assay. Immunocytochemistry was performed to assess variation of the intracellular distribution of RIZ protein on all- trans-retinoic acid treatment. Forced expression of RIZ1 protein was obtained with a recombinant adenovirus containing RIZ1 cDNA. RESULTS: Treatment with retinoic acid induced a selective expression of RIZ1 in HL60 cell line. Retinoic acid effect was maximal at 7 days and correlated to the granulocytic differentiation of cells. A similar effect was obtained in retinoic acid-sensitive NB4 cell line or in patients' leukemic promyelocytes, but not in the retinoic acid-resistant cell line NB4.007/6 or in the U937 cell line. Selective expression of RIZ1 was also induced by 12-O-tetradecanoyl-phorbol-13-acetate in the U937 and HL60 cell lines and by 1,25-dihydroxyvitamin D(3) only in HL60 cells. In HL60 cells, RIZ1 was also induced by activation of a retinoid alpha receptor-independent maturation pathway based on retinoid X receptor agonist and protein kinase A synergism. In addition, retinoic acid produced a redistribution of the antigen within the nucleus in these cells. Forced expression of RIZ1 protein induced growth arrest and death of HL60 cells. CONCLUSIONS: The correlation between the selective expression of RIZ1 induced by retinoic acid, 12-O-tetradecanoyl-phorbol-13-acetate, or 1,25-dihydroxyvitamin D(3) and differentiation suggested that RIZ protein was involved in myeloid cell differentiation induced by these agents.
Assuntos
Diferenciação Celular/fisiologia , Proteínas de Ligação a DNA , Células Mieloides/fisiologia , Proteínas Nucleares/metabolismo , Fatores de Transcrição , Monofosfato de Adenosina/análogos & derivados , Adenoviridae/metabolismo , Antineoplásicos/farmacologia , Benzoatos/farmacologia , Células Cultivadas , Histona-Lisina N-Metiltransferase , Humanos , Immunoblotting , Imuno-Histoquímica , Células Mieloides/citologia , Células Mieloides/efeitos dos fármacos , Proteínas Nucleares/genética , Proteína do Retinoblastoma/genética , Proteína do Retinoblastoma/metabolismo , Retinoides/farmacologia , Tretinoína/farmacologia , Dedos de Zinco/genéticaRESUMO
Estrogen treatment of MCF-7 cells grown in serum-free medium induced a modification of the intracellular distribution of p53 protein. Western blot analysis and immunofluorescence staining showed that p53 was localized in the nucleus of untreated cell and that after 48 h of hormone treatment, it was mostly localized in the cytoplasm. This effect was blocked by the antiestrogen ICI182,780. Intracellular redistribution of p53 was correlated to a reduced expression of the WAF1/CIP1 gene product and to the presence of degradation fragments of p53 in the cytosol. Estradiol treatment prevented the growth inhibition induced by oligonucleotide transfection, simulating DNA damage. This observation indicated that the wild-type p53 gene product present in the MCF-7 cell could be inactivated by estradiol through nuclear exclusion to permit the cyclin-dependent phosphorylation events leading to the G1-S transition. In addition, the estradiol-induced inactivation of p53 could be involved in the tumorigenesis of estrogen-dependent neoplasm.
Assuntos
Neoplasias da Mama/metabolismo , Estradiol/metabolismo , Proteína Supressora de Tumor p53/antagonistas & inibidores , Western Blotting , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/biossíntese , Dano ao DNA , Eletroforese em Gel de Poliacrilamida , Estradiol/análogos & derivados , Estradiol/farmacologia , Antagonistas de Estrogênios/farmacologia , Feminino , Fulvestranto , Fase G1 , Humanos , Imuno-Histoquímica , Fase S , Transfecção , Células Tumorais CultivadasRESUMO
Co-immunoprecipitation experiments in cell extract from cultured cells or target tissues indicated that estrogen receptor was complexed with the retinoblastoma binding protein RIZ in a ligand-dependent manner. Mapping of interaction sites indicated that in both proteins the same regions and motifs responsible for the interaction of transcriptional co-activator and nuclear receptors were involved. In cultured cells, estradiol induced a redistribution of RIZ protein within the nucleus and in the cytoplasm. A similar effect was produced in vivo, in prepuberal rat endometrium, by administration of a physiological dose of estradiol. Therefore, RIZ protein could be a specific effector of estrogen action downstream of the hormone-receptor interaction, presumably involved in proliferation control.
Assuntos
Proteínas de Ligação a DNA , Estrogênios/fisiologia , Proteínas Nucleares/metabolismo , Fatores de Transcrição , Dedos de Zinco , Sequência de Bases , Linhagem Celular , Primers do DNA , Histona-Lisina N-Metiltransferase , Humanos , Receptores de Estrogênio/metabolismoRESUMO
Double-stranded DNA fragments were selected from a random pool by repeated cycles of estrogen receptor-specific immunoprecipitation in the presence of a nuclear extract and PCR amplification (cyclic amplification and selection of target, CAST, for multiple elements). Fragments were cloned and sequence analysis indicated the 5-nucleotide word TTGGC was the most recurrent sequence unrelated to the known estrogen responsive element. Screening a HeLa cell expression library with a probe designed with multiple repeats of this sequence resulted in the identification of a 1700-aa protein showing a complete homology with the product of the human retinoblastoma-interacting zinc-finger gene RIZ. In transfection experiments, RIZ protein was able to bestow estrogen inducibility to a promoter containing an incomplete estrogen responsive element and a TTGGC motif. RIZ protein present in MCF-7 cell nuclear extract retarded the TTGGC-containing probe in an EMSA. Estrogen receptor was co-immunoprecipitated from MCF-7 cell extract by antibodies to RIZ protein and vice versa, thus indicating an existing interaction between these two proteins.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas Nucleares/genética , Receptores de Estrogênio/genética , Fatores de Transcrição , Sequência de Bases , Proteínas de Ligação a DNA/metabolismo , Células HeLa , Histona-Lisina N-Metiltransferase , Humanos , Dados de Sequência Molecular , Proteínas Nucleares/metabolismo , Receptores de Estrogênio/metabolismo , Análise de Sequência , Transfecção , Dedos de ZincoRESUMO
The mechanism by which estradiol acts on cell multiplication is still unclear. Under conditions of estradiol-dependent growth, estradiol treatment of human mammary cancer MCF-7 cells triggers rapid and transient activation of the mitogen-activated (MAP) kinases, erk-1 and erk-2, increases the active form of p21ras, tyrosine phosphorylation of Shc and p190 protein and induces association of p190 to p21ras-GAP. Both Shc and p190 are substrates of activated src and once phosphorylated, they interact with other proteins and upregulate p21ras. Estradiol activates the tyrosine kinase/p21ras/MAP-kinase pathway in MCF-7 cells with kinetics which are similar to those of peptide mitogens. It is only after introduction of the human wild-type 67 kDa estradiol receptor cDNA that Cos cells become estradiol-responsive in terms of erk-2 activity. This finding, together with the inhibition by the pure anti-estrogen ICI 182 780 of the stimulatory effect of estradiol on each step of the pathway in MCF-7 cells proves that the classic estradiol receptor is responsible for the transduction pathway activation. Transfection experiments of Cos cells with the estradiol receptor cDNA and in vitro experiments with c-src show that the estradiol receptor activates c-src and this activation requires occupancy of the receptor by hormone. Our experiments suggest that c-src is an initial and integral part of the signaling events mediated by the estradiol receptor.