Risk assessment for preterm delivery using the fetal fibronectin test associated with the measurement of uterine cervix length in symptomatic pregnant women / Avaliação do risco para parto pré-termo utilizando o teste da fibronectina fetal, associado à medida de comprimento de colo uterino, em gestantes sintomáticas

Abstract Objective To analyze the use of the measurement of uterine cervix length (MUCL) and the fetal fibronectin (fFN) rapid test as predictors of preterm delivery (PTD) in symptomatic pregnant women assisted at the Santa Casa de Misericórdia de Sobral Maternity Hospital. Methods This was a prospective and analytic study involving 53 parturients assisted between September of 2015 and July of 2016; the participants were between 24 and 34 weeks of gestational age (GA) and presented complaints related to preterm labor (PTL) prodromes. Vaginal secretion was collected for fFN testing, and the MUCL was obtained via transvaginal ultrasonography. Results A total of 58.49% of the subjects showed MUCL < 25 mm, and 41.51% were positive in the fFNrapid test.Atotal of 48 patients were followed-up until their delivery date, and 54.17% resulted in PTL. The relative risk (RR) for PTD in patients with MUCL < 25 mm was 1.83 (p = 0.09, 0.99-3.36, 95% confidence interval [CI]), with a mean time before delivery of 2.98 weeks. Based on fFN positive results, the RR was 3.50 (p = 0.002, 1.39- 8.79, 95%CI) and themean time until delivery was 1.94weeks. The RRwas 2.70 (p = 0.002, 1.08-6.72, 95%CI) when both tests were used. The RR of PTD within 48 hours, and 7 and 14 days were, respectively, 1.30 (p = 0.11, 95% CI 1.02-1.67), 1.43 (p = 0.12, 95% CI % 0.99-2.06), and 2.03 (p = 0.008, 95% CI 1.26-3.27), when based on the MUCL, and 1.75 (p = 0.0006, 95% CI 1.20-2.53), 2.88 (p = 0.0001, 95% CI, 1.57-5.31), and 3.57 (p = 0.0002, 95% CI 1.63-7.81) when based on positive fFN results. The RR at 48 hours and 7 and 14 days considering both tests was 1.74 (p = 0.0001, 95% CI 1.14-2.64), 2.22 (p = 0.0001, 95% CI 1.22-4.04), and 2.76 (p = 0.0002, 95% CI 1.27-5.96), respectively. Conclusion In symptomatic pregnant women, we concluded that the MUCL < 25 mm associated with positive fFN rapid test indicate increased the risk for PTD. Further studies with larger sample sizes could contribute in supporting the results presented in the current study.

Resumo Objetivo Analisar a utilização da medida do comprimento do colo uterino (MCCU), e do teste da fibronectina fetal (FNf) como preditores do trabalho de parto pré-termo (PPT), em gestantes sintomáticas, atendidas na Maternidade da Santa Casa de Misericórdia de Sobral. Métodos Foi realizado umestudo prospectivo e analítico, envolvendo 53 parturientes atendidas no período de setembro de 2015 a julho de 2016, com idade gestacional (IG) entre 24 e 34 semanas que tiveram queixas relacionadas a pródromos de trabalho de parto prematuro (TPP), sendo realizada coleta de secreção vaginal para FNf e MCCU por via ultrassonográfica transvaginal. Resultados Um total de 58,49% das pacientes tinham MCCU < 25 mm, e 41,51% tiveram teste rápido de fFN positivo. Foi feito o acompanhamento de 48 pacientes, com 54,17% de PPTs. O risco relativo (RR) para PPT com MCCU < 25 mm foi de 1,83 (p = 0,09, 0,99-3,36, intervalo de confiança [IC] 95%), com média de tempo até o parto de 2,98 semanas. Para fFN, o RR foi de 3.50 (p = 0.002, 1.39-8.79, IC 95%) e a média até o parto foi de 1,94 semanas. Quando os dois testes forampositivos, o RR foi de 2,70 (1,08-6,72). Para a MCCU, o RR para PPT em 48 horas, 7 e 14 dias foram 1,30 (p = 0.11, 95% IC 1.02-1.67), 1,43 (p = 0.12, 95% CI % 0.99-2.06) e 2,03 (p = 0.008, 95% IC 1.26-3.27), respectivamente. Para FNf, em 48 horas, 7 e 14 dias foi de 1,75 (p = 0.0006, 95% IC 1.20-2.53, 2,88 (p = 0.0001, 95% IC, 1.57-5.31) e 3,57 (p = 0.0002, 95% IC 1.63-7.81) respectivamente. Com os dois testes, o RR em 48 horas, 7 e 14 dias foi 1,74 (p = 0.0001, 95%IC 1.14-2.64), 2,22 (p = 0.0001, 95% IC 1.22-4.04) e 2,76 (p = 0.0002, 95% IC 1.27-5.96) respectivamente. Conclusão Em mulheres grávidas sintomáticas, concluímos que a MCCU < 25 mm e o teste rápido de FNf positivo indicam aumento do risco de PPT. Outros estudos com tamanhos de amostra maiores podem contribuir para apoiar os resultados apresentados no presente estudo.

Avances en el tratamiento de cáncer de próstata resistente a la castración: énfasis en nuevas terapias hormonales / Advances in the treatment of castration-resistant prostate cancer: emphasis in new hormonal therapies

Prostate cancer represents the second cancer-related cause of death in North American and Chilean men. The main treatment for incurable stages of disease is surgical or pharmacological castration. However, with time and despite the addition of anti-androgens, the disease progresses to a clinical state that has been commonly referred to as “hormone refractory”. In recent years, the concept of hormone refractoriness has been challenged and replaced by “castration resistance”, acknowledging that further and optimal hormonal manipulation can be attained, beyond achieving testosterone levels at castration range. The purpose of this review is to summarize the recent therapeutic breakthroughs in the management of metastatic castrate resistant prostate cancer (mCRPC), with greater emphasis in the newer hormonal therapy agents such as Abiraterone and Enzalutamide. Future combination and sequential treatment strategies are contextualized in the current era of personalized cancer medicine and genomic characterization of prostate cancer.

Rapamycin Inhibits Platelet-Derived Growth Factor- Induced Collagen, but Not Fibronectin, Synthesis in Rat Mesangial Cells

Rapamycin, a macrocyclic lactone, is effective in reducing the incidence of acute rejection after renal transplantation. The inhibitory effects of rapamycin on lymphocyte proliferation and the molecular mechanisms that were involved have been described. However, its effects on glomerular mesangial cells have not been clearly understood, and here, we examined the effect of rapamycin on platelet-derived growth factor (PDGF) - induced extracellular matrix synthesis as well as cell proliferation in mesangial cells. Rat mesangial cells were isolated from the glomeruli of Sprague-Dawley rats and cultured with Dulbecco's modified Eagles medium containing 20% fetal bovine serum. Different concentrations of rapamycin were administered 1 hour before the addition of 10 ng/ml of PDGF into growth arrested and synchronized cells. Cell proliferation was assessed by [3H]thymidine incorporation, total collagen synthesis by [3H]proline incorporation, and fibronectin secretion into the medium by Western blot analysis. In the mesangial cells, PDGF increased cell proliferation by 4.6-fold, total collagen synthesis by 1.8-fold, and fibronectin secretion by 3.2-fold. Rapamycin above 10 nM significantly inhibited PDGF-induced proliferation and collagen synthesis, but the treatment of rapamycin up to 1micrometer did not show any significant effects on PDGF-induced fibronectin secretion. These inhibitory effects of rapamycin on PDGF-induced mesangial cell proliferation and collagen synthesis reflect the potential value of rapamycin in the prevention and treatment of glomerulosclerosis in patients with chronic allograft nephropathy.

Molecular cloning of recombinant fibronectin EDA and EDB fusion protein / 法医学杂志

OBJECTIVE@#Construct a recombinant plasmid pET28a-EDA-EDB, prepare the fusion EDA-EDB protein.@*METHODS@#For the production of recombinant fibronectin EDA-EDB in Escherichia coli, the EDA and EDB segments were separated from pGEM2-EDA/EDB and recomposed with two additional amino acids, then cloned into the expression vector pET28a. pET system to express EDA-EDB fusion protein and 6 x His/Ni-NTA system to purify it in a single step were used. Western blotting confirmed the purified protein.@*RESULTS@#The EDA and EDB segments were ligated and inserted into pET28a vector. EDA-EDB fusion protein was highly expressed in Escherichia coli BL21 (DE3). Afterwards, it was purified by Ni-NTA resin and verified by western blotting.@*CONCLUSION@#EDA-EDB fusion protein can be expressed in pET system and purified by 6 x His/Ni-NTA system.

Efeito da desnutriçäo protéica sobre a matriz extracelular da medula óssea de camundongos / Effect of the protein malnutrition on the extracellular matrix of the bone marrow of mices

As células sangüíneas originam-se da medula óssea através da célula tronco que sofre processo de proliferação, diferenciação e maturação no microambiente hematopoiético. O microambiente hematopoiético é uma estrutura altamente organizada composta de células estromais, moléculas da matriz extracelular (MEC) e citocinas. A desnutrição protéico-energética diminui a produção de células sangüíneas e interfere na defesa do organismo. Neste trabalho estudamos os efeitos da desnutrição protéica (dieta contendo 4 por cento de caseína) sobre a MEC da medula óssea em camundongos. Avaliamos a composição da MEC através de SDS-PAGE 7,5 por cento e Western blot para fibronectina (FN), laminina (LN) e trombospondina (TSP)...

Chronic liver injury, TGF-beta, and cancer

Cells termed myofibroblasts are prominent in the injury response of all epithelial tissues. They exhibit proliferation, migration, production of collagen and other extracellular matrix (ECM) molecules, and contraction, all for containing the injury and closing the wound. When the injury is limited in time, the final stage of the repair involves a dismantling of the cellular apparatus and restoration of normal tissue structure. With multiple cycles of repair, however, there is net accumulation of ECM, to the detriment of tissue structure and function. Repair-related ECM coalesces into fibrous bundles and, over time, undergoes changes that render it resistant to degradation. The result is a scar. In the skin, a scar may have cosmetic importance only. In the liver, however, extensive scarring is the setting for unregulated growth and neoplasia; also, fibrous bands disrupt normal blood flow, leading to portal hypertension and its complications. With regard to therapy for fibrosis, the first consideration is elimination of the injury factor. However, given that many liver diseases do not have effective therapies at present, strategies targeting fibrogenesis per se are under development. The main source of myofibroblast-like cells and ECM production in the liver is the perisinusoidal stellate cell, which responds to injury with a pleiotypic change termed activation. Activation is orchestrated by cytokines and the ECM itself. Among the cytokines involved in this process, transforming growth factor-beta (TGF-beta) is particularly prominent. The early changes in ECM include de novo production of a specific "fetal" isoform of fibronectin, which arises from sinusoidal endothelial cells. It is stimulated by TGF-beta and acts directly on stellate cells to promote their activation. Based on these and other advances in understanding the fundamentals of the injury response, several strategies now exist for altering fibrogenesis, ranging from agents that block TGF-beta to traditional Chinese herbal extracts. Arrest of fibrogenesis, even with underlying cirrhosis, is likely to extend life or prolong the time to transplant. Whether it reduces the risk of hepatocellular carcinoma remains to be proven. Although TGF-beta antagonists are effective anti-fibrogenic agents, they will require detailed safety testing because of the finding that several forms of epithelial neoplasia are associated with altered regulation of TGF-beta.

Study on the expression of fibronectin after cerebral contusion in rats for timing of injuries / 法医学杂志

An experimental model of reproducible focal cerebral contusions in rats was made by a free-drop impacting right hemisphere. The expression of fibronectin and its mRNA after cerebral contusion were detected respectively by immunohistochemical staining and in situ hybridization. Results indicated that the expression of fibronectin and its mRNA increased after injury, and there existed a relationship between increased fibronectin and its mRNA and different intervals after brain injury. It is inferred that the expression of fibronectin and its mRNA can be used for timing of brain injuries and distinguishing antemortem and postmortem brain contusions.

Fibronectin in the trabecular meshwork: immunofluorescent and immunoelectron microscopic findings

We examined the fibronectin (FN) secretion of cultured trabecular meshwork (TM) cells in a normal human eye by indirect immunofluorescent technique using mouse anti-human FN monoclonal antibody and FITC-conjugated goat anti-mouse IgG. To localize FN on frozen sections of normal TM, which were obtained from 7 enucleated eyes owing to traumatic eyeball rupture, the same indirect immunofluorescent method was used. Immunoelectron microscopy was applied to demonstrate the distribution pattern of FN in the normal TM of 2 human eyes using an avidin-biotin-peroxidase complex method. In the tissue culture of TM, the TM cell walls and extracellular matrices showed an intense staining with antibody to FN. Indirect immunofluorescent staining of FN on frozen sections of TM showed strong positive reactions in the subendothelial region. There was no reaction in the central core of the trabecular beam. Immunoelectron microscopy revealed the reaction products to FN in the areas lining the trabecular endothelial cells.

Fibronectina: respuesta clínica a su corrección en pacientes traumatizados y sépticos

Se estudio un grupo de 15 pacientes con patología traumática y/o séptica, en quienes se dosificaron los niveles iniciales de fibronectina (FN), encontrando cifras subnormales en todos los casos (X: 212 mcgr/ml). Posteriormente, cuando las condiciones clínicas empezaron a deteriorarse con alteraciones hemodinámicas importantes y falla respiratoria y/o neurológica, se administraron 10 unidades de criprecipitado del plasma, como suuplemento de su esquema terapéutico básico. Se obtuvo elevación de los niveles iniciales de FN hasta 306 mcgr/ml (p<0.02), a las dos horas de aplicación del crioprecipitado y normalización de éstos al séptimo día. Se correlacionaron los niveles bajos e FN con leucocitosis y aumento de cayados, cifras que al normalizarse coinciden con la elevación de FN y la mejoría del estado clínico. La mortalidad en este grupo fue de 21.4 por ciento. Nueve pacientes con patologías similares e idénticas medidas terapéuticas se tomaron como control sin administrarles crioprecipitados. La mortalidad fue del 78.6 por ciento. Reemplazar las perdidas de FN puede ser una nueva modalidad terapéutica, que amerita estudios aleatorios adecuados con el fin de establecer su verdadero valor en el manejo de los pacientes sépticos y traumatizados

Efecto quimiotáctico de los sobrenadantes de cultivos de glomerulos de rata normal sobre macrófagos peritoneales / Chemotactic effect of the supernatants of normal rats glomerular cultures on peritoneal macrophages

En estudios previos se ha determinado que el mesangio glomerular es repoblado periodicamente por células LC+ e Ia+ procedentes de la médula ósea. El presente estudio fue realizado con el objeto de determinar si el glomerulo normal de rata era capaz de producir substancias con capacidad de atraer células (quimiotaxis) provenientes de la médula ósea (macrófagos) y de esta manera acondicionar la penetración en el mesangio de estas células. Para los efectos, se estudiaron los sobrenadantes de cultivos de glomérulos provenientes de ratas normales; encontrándose efecto quiomiotáctico sobre macrófagos obtenidos de la cavidad peritonial de la rata. A su vez, con el objeto de identificar a las posibles substancias que pudiesen causar el efecto quimiotáctico se analizaron en los sobrenadantes la presencia de Fibronectina e Interleucina 1 (ambas substancias quimiotácticas para células del sistema monocito/macrofago). En todos los sobrenadantes se apreció la presencia de ambas substancias. Se concluye de estos resultados que la repoblación del mesangio por parte de células del sistema monocito/macrofago puede ser debida a la presencia de substancias quimiotácticas producidas por el glomérulo y que estas substancias podrían ser la Fibronectina, la Interleucina 1 o ambas