Evaluation of the antitumoral effect of dihydrocucurbitacin-B in both in vitro and in vivo models

Aims  We evaluated both in vitro and in vivo antitumoral properties of an isolated compound from Wilbrandia ebracteata, dihydrocucurbitacin-B (DHCB), using B16F10 cells (murine melanoma). Materials and methods  We made use of MTT and ³H-Thymidine assays to investigate the cell viability and cell proliferation, flow cytometry analysis to monitor cell cycle and apoptosis, western blot analysis to evaluate the expression of cell cycle proteins, imunofluorescence analysis and in vivo tumor growth and metastasis. Results  Dihydrocucurbitacin-B significantly reduced cell proliferation without important effects on cells viability. DHCB lead cells to accumulate in G2/M phases accompanied by the appearance of polyploid cells, confirmed by fluorescence assays that demonstrated a remarkable alteration in the cell cytoskeleton and formation of binuclear cells. Annexin-V-FITC incorporation demonstrated that DHCB did not induce apoptosis. About 10 μg/mL DHCB was found to decrease cyclin-A, and especially in cyclin-B1. The in vivo experiments showed that DHCB treatment (once a day up to 12 days; p.o.) was able to reduce the tumor growth and lung metastasis up to 83.5 and 50.3%, respectively. Conclusions  Dihydrocucurbitacin-B reduces cell proliferation due to a decrease in the expression of cyclins, mainly cyclin-B1 and disruption of the actin cytoskeleton, arresting B16F10 cells in G2/M phase. Taken together, the in vitro and in vivo experiments suggest that DHCB was effective against cancer, however, it remains to be proved if DHCB will be a good candidate for drug development.     

Association of marital status with takotsubo syndrome (broken heart syndrome) among adults in the United States

BACKGROUNDThe pathophysiology of takotsubo syndrome (TTS) is not well understood, however, it is often precipitated by psychological or physical stress. Marital status is related to emotional stress, but its associations with TTS are limited.AIMTo explored the potential association between marital status and TTS. METHODSWe conducted a case-control study using data on patients aged ≥ 40 years with marital status data in the National Hospital Discharge Survey (2006-2010). The International Classification of Diseases Ninth Revision codes were used to identify cases with TTS and other comorbid conditions. Each case was matched to 5 controls by age, sex, year of TTS diagnosis and bed size of hospital. Two sets of controls were selected: Acute myocardial infarction (AMI) controls and non-cardiovascular disease (CVD) controls. Conditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) for the association of marital status with TTS. RESULTSThe 59 patients with TTS who had information on marital status were matched to 295 controls with AMI and 295 non-CVD controls, resulting in a sample of 649 patients. The average age of cases was 69.7 ± 11 years with 90% being women and 88% reporting White race. In multivariable-adjusted models, compared to singles, patients who were married had lower odds of TTS (OR = 0.86, 95%CI: 0.79–0.93) while those who were widowed (OR = 1.14, 95%CI: 1.05–1.23) or divorced/separated (OR = 1.32, 95%CI: 1.21–1.45) had elevated odds for TTS when compared to non-CVD controls. Similar results were observed when cases were compared to controls with AMI. CONCLUSIONIn this study, being married was associated with lower odds for TTS while being divorced/separated or widowed was related to elevated odds for TTS. These novel findings that underscore the potential importance of social factors like marital status in the development of TTS need confirmation in larger studies.     

Study of the Mutagenic Activity of Extracts of the Plant Wilbrandia ebracteata

The present study was carried out to investigate the mutagenic activity of six extracts of the plant Wilbrandia ebracteata (family Cucurbitacea), one of the species found in products commercially sold as "Taiuiá'. The method for investigation was the Salmonella/microsome assay using strains TA100, TA98 and TA102 in assays carried out in the presence or absence of S9Mix as the activating system. The results indicate the absence of mutagenic activity in flavonoid and cucurbitacin-rich fractions. Signs of an increased mutagenic index were detected in the methanol fractions prepared from dried roots. We observed an increased dose response below a revertant rate, which was twice the spontaneous yield.     

Development of a caffeic acid–phthalimide hybrid compound for NADPH oxidase inhibition

NADPH oxidases are pharmacological targets for the treatment of inflammation-based diseases. This work presents the synthesis and study of a caffeic acid/phthalimide hybrid compound (C2) as a potential inhibitor of NADPH oxidases. Throughout the study, we have compared compound C2 with its precursor caffeic acid (C1). The redox properties were compared using three different antioxidant methodologies and showed that C2 was slightly less effective than C1, a well-established and robust antioxidant. However, C2 was three-fold more effective than albumin (used as a model protein). This chemical feature was decisive for the higher efficiency of C2 as an inhibitor of the release of superoxide anions by stimulated neutrophils and enzymatic activity of cell-free NADPH oxidase. Docking simulation studies were performed using the crystal structure of the recombinant dehydrogenase domain of the isoform NOX5 of C. stagnale, which retains the FAD cofactor (PDB: 5O0X). Considering that C2 could bind at the FAD redox site of NOX5, studies were conducted by comparing the interactions and binding energies of C1 and C2. The binding energies were −50.30 (C1) and −74.88 (C2) (kJ mol⁻¹), which is in agreement with the higher efficacy of the latter as an NADPH oxidase inhibitor. In conclusion, incorporating the phthalimide moiety into caffeic acid was decisive for its effectiveness as an NADPH oxidase inhibitor.

The incorporation of the phthalimide moiety into caffeic acid was decisive for its effectiveness as an NADPH oxidase inhibitor.     

Maternal C-reactive protein concentrations during pregnancy and birth weight in a prospective cohort in Rio de Janeiro,Brazil

Objective: To evaluate the association between maternal C-reactive protein (CRP) concentrations during pregnancy and birth weight (BW) Z-score.

Methods: A prospective cohort of pregnant women were followed at 5–13 (n?=?203), 20–26 (n?=?181), and 30–36 (n?=?181) gestational weeks and at 30–45 d postpartum. Maternal CRP concentrations were assessed three times during pregnancy using immunoturbidimetric methods (ultra-sensitive kits). BW Z-score and newborns classified as small for gestational age (SGA) were evaluated according to Intergrowth-21st curves. Statistical analyses included SGA rates, BW Z-score means (SD) and a two-stage procedure: (1) a linear mixed-effect model (LME) to predict CRP intercept (mean exposure level) and slope (trend of change during pregnancy); and (2) a multiple linear regression model with BW Z-score as the outcome and CRP intercept and slope exposures.

Results: A total of 4.4% (n?=?9) women delivered SGA newborns. The mean BW was 3282.0 (37.3) g, and the mean gestational age at delivery was 38.8 (0.1) weeks. Women in the third tertile of the CRP rate of change gave birth to infants with a mean BW Z-score that was lower than those in the first/second tertiles (0.226 versus 0.381; p?=?0.324). For the adjusted baseline CRP (β?=?0.08; 95% CI: 0.03–0.14), the CRP trend of change was inversely associated with the BW Z-score (β=??3.77; 95% CI: ?5.45 to ?2.10).

Conclusions: The maternal CRP trend of change during pregnancy was negatively associated with BW Z-score.     

人体脂肪基质细胞的分离培养及其成骨活性的研究

目的：脂肪组织来源的脂肪基质细胞具备向其他多种组织成分，如神经细胞、骨骼肌细胞、心肌细胞、软骨细胞、骨细胞等分化的能力。探讨人体脂肪组织来源脂肪基质细胞的分离培养方法及其成骨活性。方法：于2005—10/2006-05选取临床手术中收集的废弃脂肪组织及行脂肪抽吸术所得的脂肪组织为实验材料．采用机械切割及I型胶原酶消化法从脂肪组织中分离获得脂肪基质细胞．原代培养后，以诱导培养基（内含10mmol／Lβ-甘油磷酸钠、10^-8mol／L地塞米松、50mg／L维生素C）进行诱导培养。实验评估：应用形态学观察、碱性磷酸酶钙一钴法染色检测碱性磷酸酶，VonKossa钙化结节染色检测钙化结节的形成等方法来鉴定诱导分化所得细胞的成骨活性。结果：行诱导培养的脂肪基质细胞呈多层成长，形态多为梭形、锥形或立方形，细胞外基质有白色钙化结节形成；细胞增殖速度减慢，生长周期变长。碱性磷酸酶钙钴法染色及VonKossa钙化结节染色后均表现为阳性，未诱导培养的脂肪基质细胞则表现为阴性。结论：初步建立了一套由脂肪组织分离培养脂肪基质细胞的方法，并证明该细胞具备成体干细胞的特性，在体外诱导培养条件下具备向成骨细胞分化的能力。