Quantitative Ultrasound in Ex Vivo Fibrotic Rabbit Livers.

Liver fibrosis is the common result of chronic liver disease. Diagnosis and grading liver fibrosis for patient management is mainly based on blood tests and hepatic puncture-biopsy, which is particularly invasive. Quantitative ultrasound (QUS) techniques provide insight into tissue microstructure and are based on the frequency-based analysis of the signals from biologic tissues. This study aims to quantify how spectral-based QUS parameters change with fibrosis grade. The changes in QUS parameters of healthy and fibrotic rabbit liver samples were investigated and were compared with the changes in liver stiffness, using shear wave elastography. Overall, the acoustic concentration was found to decrease with increasing fibrosis grade, and the effective scatterer size was found to be higher in fibrotic livers when compared with normal liver. The result of this study indicates that the combination of three QUS parameters (stiffness, effective scatterer size and acoustic concentration) provides the best classification performance, especially for classifying healthy and fibrotic livers.

Development of a Fluid Dynamic Model for Quantitative Contrast-Enhanced Ultrasound Imaging.

Contrast-enhanced ultrasound (CEUS) is a non-invasive imaging technique extensively used for blood perfusion imaging of various organs. This modality is based on the acoustic detection of gas-filled microbubble contrast agents used as intravascular flow tracers. Recent efforts aim at quantifying parameters related to the enhancement in the vascular compartment using time-intensity curve (TIC), and at using these latter as indicators for several pathological conditions. However, this quantification is mainly hampered by two reasons: first, the quantification intrinsically solely relies on temporal intensity variation, the explicit spatial transport of the contrast agent being left out. Second, the exact relationship between the acquired US-signal and the local microbubble concentration is hardly accessible. This paper introduces the use of a fluid dynamic model for the analysis of dynamic CEUS (DCEUS), in order to circumvent the two above-mentioned limitations. A new kinetic analysis is proposed in order to quantify the velocity amplitude of the bolus arrival. The efficiency of proposed methodology is evaluated both in-vitro, for the quantitative estimation of microbubble flow rates, and in-vivo, for the classification of placental insufficiency (control versus ligature) of pregnant rats from DCEUS. Besides, for the in-vivo experimental setup, we demonstrated that the proposed approach outperforms the performance of existing TIC-based methods.

Effect of myricetin, pyrogallol, and phloroglucinol on yeast resistance to oxidative stress.

The health beneficial effects of dietary polyphenols have been attributed to their intrinsic antioxidant activity, which depends on the structure of the compound and number of hydroxyl groups. In this study, the protective effects of pyrogallol, phloroglucinol, and myricetin on the yeast Saccharomyces cerevisiae were investigated. Pyrogallol and myricetin, which have a pyrogallol structure in the B ring, increased H2O2 resistance associated with a reduction in intracellular oxidation and protein carbonylation, whereas phloroglucinol did not exert protective effects. The acquisition of oxidative stress resistance in cells pretreated with pyrogallol and myricetin was not associated with an induction of endogenous antioxidant defences as assessed by the analysis of superoxide dismutase and catalase activities. However, myricetin, which provided greater stress resistance, prevented H2O2-induced glutathione oxidation. Moreover, myricetin increased the chronological lifespan of yeast lacking the mitochondrial superoxide dismutase (Sod2p), which exhibited a premature aging phenotype and oxidative stress sensitivity. These findings show that the presence of hydroxyl groups in the ortho position of the B ring in pyrogallol and myricetin contributes to the antioxidant protection afforded by these compounds. In addition, myricetin may alleviate aging-induced oxidative stress, particularly when redox homeostasis is compromised due to downregulation of endogenous defences present in mitochondria.

Non-menopausal Status, High Nuclear Grade, Tumor Size >30 mm and Positive Resection Margins Are Predictors of Residual Tumor After Lumpectomy for Ductal Carcinoma In Situ of the Breast.

AIM: Breast-conserving surgery with radiation therapy is the primary treatment for ductal carcinoma in situ (DCIS). Re-excision is indicated when clear resection margins have not been achieved, although in some cases the procedure may be unnecessary as there is no residual tumor. The purpose of our three-Center retrospective study was to identify predictors of positive re-excision findings following breast-conserving surgery for DCIS. PATIENTS AND METHODS: A total of 285 patients underwent re-excision following conservative treatment for DCIS between 01/01/08 and 12/31/13 at three breast-cancer referral Centers. We conducted a retrospective, comparative review of the factors that differentiated patients with a residual tumor from those without. The study was based on clinical, radiological, surgical and pathological criteria. RESULTS: A total of 180 patients (63%) had residual tumor after conservative treatment. Six factors were predictive on univariate analysis: young age (p=0.025), non-menopausal status (p=0.016), absence of preoperative biopsy (p=0.0029), high nuclear grade (p=0.0181), lesion size >30 mm (p=0.032), and positive surgical margins (p=0.0016). Four factors remained independently predictive on multivariate analysis: non-menopausal status (p=0.0017), high nuclear grade (p=0.0031), lesion size >30 mm (p=0.012) and positive surgical margins (p=0.0013). We calculated a 93% probability of positive re-excision findings if all four factors were combined. On the other hand, if none of the factors were present, the rate fell to 18%. CONCLUSION: In cases of DCIS, where risk factors for both involved lumpectomy margins and recurrence are carefully studied, knowledge of the risk factors for residual tumor can help guide therapeutic choices.

Quercetin protects Saccharomyces cerevisiae against oxidative stress by inducing trehalose biosynthesis and the cell wall integrity pathway.

BACKGROUND: Quercetin is a naturally occurring flavonol with antioxidant, anticancer and anti-ageing properties. In this study we aimed to identify genes differentially expressed in yeast cells treated with quercetin and its role in oxidative stress protection. METHODS: A microarray analysis was performed to characterize changes in the transcriptome and the expression of selected genes was validated by RT-qPCR. Biological processes significantly affected were identified by using the FUNSPEC software and their relevance in H(2)O(2) resistance induced by quercetin was assessed. RESULTS: Genes associated with RNA metabolism and ribosome biogenesis were down regulated in cells treated with quercetin, whereas genes associated with carbohydrate metabolism, endocytosis and vacuolar proteolysis were up regulated. The induction of genes related to the metabolism of energy reserves, leading to the accumulation of the stress protectant disaccharide trehalose, and the activation of the cell wall integrity pathway play a key role in oxidative stress resistance induced by quercetin. CONCLUSIONS: These results suggest that quercetin may act as a modulator of cell signaling pathways related to carbohydrate metabolism and cell integrity to exert its protective effects against oxidative stress.

The high frequency of sperm aneuploidy in klinefelter patients and in nonobstructive azoospermia is due to meiotic errors in euploid spermatocytes.

For nonobstructive azoospermic (NOA) patients with a normal karyotype or for Klinefelter syndrome (47,XXY) patients, intracytoplasmic sperm injection is associated with an increased aneuploidy risk in offspring. We examined testicular cells from patients with different azoospermia etiologies to determine the origin of the aneuploid spermatozoa. The incidence of chromosome abnormalities was investigated in all types of azoospermia. Four study subgroups were constituted: Klinefelter patients (group 1), NOA patients with spermatogenesis failure but a normal karyotype (group 2), obstructive azoospermic patients with normal spermatogenesis (group 3), and control patients with normal sperm (group 4). The pachytene stage (in the three azoospermic groups) and postmeiotic cells (in all groups) were analyzed with fluorescence in situ hybridization. No aneuploid pachytene spermatocytes were observed. Postmeiotic aneuploidy rates were higher in the two groups with spermatogenesis failure (5.3% and 4.0% for groups 1 and 2, respectively) than in patients with normal spermatogenesis (0.6% for group 3 and group 4). Whatever the etiology of the azoospermia, the spermatozoa originated from euploid pachytene spermatocytes. These results strengthen the hypothesis whereby sperm aneuploidy in both Klinefelter patients and NOA patients with a normal karyotype results from meiotic abnormalities and not from aneuploid spermatocytes. The fact that sperm aneuploidy was more frequent when spermatogenesis was altered suggests a deleterious testicular environment. The study results also provide arguments for offering preimplantation genetic diagnosis or prenatal diagnosis when a pregnancy occurs for fathers with NOA (whatever the karyotype).

Activation of the Hog1p kinase in Isc1p-deficient yeast cells is associated with mitochondrial dysfunction, oxidative stress sensitivity and premature aging.

The Saccharomyces cerevisiae Isc1p, an orthologue of mammalian neutral sphingomyelinase 2, plays a key role in mitochondrial function, oxidative stress resistance and chronological lifespan. Isc1p functions upstream of the ceramide-activated protein phosphatase Sit4p through the modulation of ceramide levels. Here, we show that both ceramide and loss of Isc1p lead to the activation of Hog1p, the MAPK of the high osmolarity glycerol (HOG) pathway that is functionally related to mammalian p38 and JNK. The hydrogen peroxide sensitivity and premature aging of isc1Δ cells was partially suppressed by HOG1 deletion. Notably, Hog1p activation mediated the mitochondrial dysfunction and catalase A deficiency associated with oxidative stress sensitivity and premature aging of isc1Δ cells. Downstream of Hog1p, Isc1p deficiency activated the cell wall integrity (CWI) pathway. Deletion of the SLT2 gene, which encodes for the MAPK of the CWI pathway, was lethal in isc1Δ cells and this mutant strain was hypersensitive to cell wall stress. However, the phenotypes of isc1Δ cells were not associated with cell wall defects. Our findings support a role for Hog1p in the regulation of mitochondrial function and suggest that constitutive activation of Hog1p is deleterious for isc1Δ cells under oxidative stress conditions and during chronological aging.

Human cancer cell antiproliferative and antioxidant activities of Juglans regia L.

Several studies suggest that regular consumption of nuts, mostly walnuts, may have beneficial effects against oxidative stress mediated diseases such as cardiovascular disease and cancer. Walnuts contain several phenolic compounds which are thought to contribute to their biological properties. The present study reports the total phenolic contents and antioxidant properties of methanolic and petroleum ether extracts obtained from walnut (Juglans regia L.) seed, green husk and leaf. The total phenolic contents were determined by the Folin-Ciocalteu method and the antioxidant activities assessed by the ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. Methanolic seed extract presented the highest total phenolic content (116 mg GAE/g of extract) and DPPH scavenging activity (EC(50) of 0.143 mg/mL), followed by leaf and green husk. In petroleum ether extracts, antioxidant action was much lower or absent. Under the oxidative action of AAPH, all methanolic extracts significantly protected the erythrocyte membrane from hemolysis in a time- and concentration-dependent manner, although leaf extract inhibitory efficiency was much stronger (IC(50) of 0.060 mg/mL) than that observed for green husks and seeds (IC(50) of 0.127 and 0.121 mg/mL, respectively). Walnut methanolic extracts were also assayed for their antiproliferative effectiveness using human renal cancer cell lines A-498 and 769-P and the colon cancer cell line Caco-2. All extracts showed concentration-dependent growth inhibition toward human kidney and colon cancer cells. Concerning A-498 renal cancer cells, all extracts exhibited similar growth inhibition activity (IC(50) values between 0.226 and 0.291 mg/mL), while for both 769-P renal and Caco-2 colon cancer cells, walnut leaf extract showed a higher antiproliferative efficiency (IC(50) values of 0.352 and 0.229 mg/mL, respectively) than green husk or seed extracts. The results obtained herein strongly indicate that walnut tree constitute an excellent source of effective natural antioxidants and chemopreventive agents.

Xanthohumol influences preadipocyte differentiation: implication of antiproliferative and apoptotic effects.

There is interest in the research of natural compounds that may interfere with the adipocyte life cycle, due to the growing prevalence of obesity and related complications. We aimed at studying the effect of xanthohumol (XN), a Humulus lupulus L. prenylflavonoid, on adipocytes measuring differentiation, proliferation, and apoptosis in 3T3-L1 cells. XN reduced differentiation, as revealed by decreased lipid content and peroxisome proliferator-activated receptor gamma expression, an effect more pronounced when cells were treated before or during differentiation induction. XN also decreased proliferation, as measured by sulforhodamine staining (IC(50) between 26 and 12 microM for 24, 48, and 72 h), and preadipocyte Ki67 expression. Apoptosis was increased in preadipocytes and adipocytes. NF-kappaB activity was stimulated by XN in preadipocytes. Results suggest that XN may reduce adipocyte number, contributing to adipocyte hypertrophy. Taking into consideration the consequences of adipocyte hypertrophy, XN does not seem to improve the metabolic profile associated with obesity.