Enzyme-Assisted Extraction of Bioactive Compounds from Raspberry (Rubus idaeus L.) Pomace.

Industrial processing of raspberries into juice and jam results in the production of with high content of lipophilic and hydrophilic phytochemicals. Usually considered as waste, raspberry pomace is occasionally cold-pressed to recover specialty oil. However, the resulting pomace press-cake (PPC) still contains 30% to 35% of lipophilic compounds, such as essential fatty acids, tocols, phytosterols, and ellagitannins initially present in pomace. In a perspective of sustainable development, we investigate an eco-friendly process using an aqueous enzyme-assisted extraction (AEAE) to simultaneously and effectively recover lipophilic compounds and polyphenols from the PPC. The performance of different combinations of carbohydrases and proteases was compared. After selecting the best enzymatic system, a definitive screening design involving six factors was then implemented to optimize the process. Under optimized conditions, 1.2 units of thermostable alkaline protease/100 g PPC, pH 9, 60 °C, and 2 hr hydrolysis, more than 38% of total PPC lipophilic content were recovered in the aqueous medium. The recovery of polyphenols and antioxidant activity was, respectively, 48% and 25% higher than obtained by extraction with methanol/acetone/water mixture. Such an AEAE extract might prove useful in food and nutraceutical applications. PRACTICAL APPLICATION: Raspberry pomace, a food industrial by-product, is often considered as waste. However, it is a rich source of phytochemicals, such as tocols, polyphenols, and polyunsaturated fatty acids. To overcome the drawbacks of organic solvent use, an enzyme-assisted extraction process was developed as an eco-friendly alternative to recover these bioactive compounds. Definitive screening design experimental design was used to enhance polyphenols and lipophilics extraction yields while reducing process costs. This extract is an oil-in-water emulsion, with high content in antioxidant phytochemicals, which might have potential for use in nutraceutical applications. Therefore, this green process developed for the valorization of raspberry pomace is considered as a perspective of sustainable development.

Pluripotent and lineage-committed CD34+ subsets in leukapheresis products mobilized by G-CSF, GM-CSF vs. a combination of both.

To define an optimal regimen for mobilizing blood-derived progenitor cells from healthy donors for allogeneic transplantation, we have studied the early and lineage-committed CD34+ subsets in the leukapheresis products after mobilization with G-CSF (10 micrograms/kg/d), GM-CSF (10 micrograms/kg/d), and the combination of G-CSF and GM-CSF (G/GM, 5 micrograms/kg/d of each). We used three color and five dimensional flow cytometry with a panel of monoclonal antibodies against CD3, CD7, CD10, CD11b, CD15, CD33, CD34, CD38, CD45, CD61, and CD71. As reference, we also analyzed CD34+ subsets in samples from umbilical cord blood (UCB) and from adult bone marrow (BM). The level of total CD34+ cells was 0.04 +/- 0.03% (mean +/- SD) in peripheral blood at baseline, and reached a maximum on day 5 or day 6 of administration of growth factors. The percentages of CD34+ cells in the leukapheresis products were 1.06 +/- 0.37% (mean +/- SD) with G-CSF mobilization, 0.35 +/- 0.24% with GM-CSF, and 0.92 +/- 0.61% with the combination of both. Among the CD34+ subsets, the percentage of cells that were CD34+/CD38- was highest in UCB (7.18 +/- 5.58%) and lowest in G-CSF mobilized peripheral blood (0.80 +/- 0.22%), whereas GM-CSF or G/GM mobilized products gave rise to intermediate levels (4.43 +/- 3.40%, 3.61 +/- 2.42%, respectively). The differences between G/GM and G-CSF, between UCB and G-CSF, or between UCB and BM are significant. The absolute numbers of CD34+/CD38- and CD34+/CD38-/HLA-DR+ subsets are also significantly higher in the G/GM mobilized products than in G-CSF products. The cloning efficiency of G/GM mobilized CD34+ cells was 2 times higher than that of G-CSF mobilized CD34+ cells, albeit the difference was statistically marginal. The profile of CD34+ subsets mobilized by the combination of G/GM approaches that found in UCB. Our data illustrate that different growth factors and regimens can preferentially mobilize different CD34+ subsets from normal donors, and that the combination of G-CSF and GM-CSF might be an optimal regimen.

Parkin maintains mitochondrial levels of the protective Parkinson's disease-related enzyme 17-ß hydroxysteroid dehydrogenase type 10.

Mutations of the PARK2 and PINK1 genes, encoding the cytosolic E3 ubiquitin-protein ligase Parkin and the mitochondrial serine/threonine kinase PINK1, respectively, cause autosomal recessive early-onset Parkinson's disease (PD). Parkin and PINK1 cooperate in a biochemical mitochondrial quality control pathway regulating mitochondrial morphology, dynamics and clearance. This study identifies the multifunctional PD-related mitochondrial matrix enzyme 17-ß hydroxysteroid dehydrogenase type 10 (HSD17B10) as a new Parkin substrate. Parkin overproduction in cells increased mitochondrial HSD17B10 abundance by a mechanism involving ubiquitin chain extension, whereas PARK2 downregulation or deficiency caused mitochondrial HSD17B10 depletion in cells and mice. HSD17B10 levels were also found to be low in the brains of PD patients with PARK2 mutations. Confocal and Förster resonance energy transfer (FRET) microscopy revealed that HSD17B10 recruited Parkin to the translocase of the outer membrane (TOM), close to PINK1, both in functional mitochondria and after the collapse of mitochondrial membrane potential (ΔΨm). PD-causing PARK2 mutations impaired interaction with HSD17B10 and the HSD17B10-dependent mitochondrial translocation of Parkin. HSD17B10 overproduction promoted mitochondrial elongation and mitigated CCCP-induced mitochondrial degradation independently of enzymatic activity. These effects were abolished by overproduction of the fission-promiting dynamin-related protein 1 (Drp1). By contrast, siRNA-mediated HSD17B10 silencing enhanced mitochondrial fission and mitophagy. These findings suggest that the maintenance of appropriate mitochondrial HSD17B10 levels is one of the mechanisms by which Parkin preserves mitochondrial quality. The loss of this protective mechanism may contribute to mitochondrial dysfunction and neuronal degeneration in autosomal recessive PD.

[Comprehension of food label claims by university undergraduate students]. / Compréhension des allégations alimentaires par des étudiants du premier cycle universitaire.

The purpose of this study was to determine whether claims made on food labels are well or poorly understood and whether the sex and the experience of consumers in buying food products influence their understanding of the various terms used in the food industry. A questionnaire was randomly distributed to 285 students registered in various undergraduate programs at the University of Ottawa. The average grade of all the respondents to the questionnaire was 53.6% which indicates a lack of understanding of the meanings of the various terms used in the food industry. The results also indicate that more frequent exposure to advertising by the food industry is apt to confuse the consumer. Finally, the respondents without experience in buying food products answered significantly better on three questions; in general, men without experience had significantly better results than women on some questions.

Simultaneous dielectric monitoring of microfluidic channels at microwaves utilizing a metamaterial transmission line structure.

The paper presents a technique that allows the simultaneous monitoring of the dielectric properties of liquids in microfluidic channels at microwave frequencies. It is capable of being integrated within the lab-on-a-chip concept and uses a composite right/left-handed transmission line resonator which is detuned by the dielectric loading of the liquids in the channels. By monitoring the change in the resonance spectrum of the resonator the loading profile can be derived with the multi-resonant perturbation method. From the value of the dielectric constant inference on the substances like cells or chemicals in the channels can be drawn. The paper presents concept, design, fabrication and characterization of prototype sensors. The sensors have been designed to operate between 20 and 30 GHz and were tested with water and water ethanol mixtures.

Reducing air-pollution: a new argument for getting drivers to abide by the speed limit?

Speeding is one of the main factors of car crash-risk, but it also contributes to increasing air-pollution. In two studies we attempted to lead drivers to abide by speed limits using "reducing air-pollution" as a new argument. We presented prevention messages that highlighted the role of speeding in increasing "crash-risk", "air-pollution", or both (Studies 1 and 2). The messages were also positively or negatively framed (Study 2). Given that women are more concerned with environmental issues than are men, we expected the following hypotheses to be validated for women. The message with the "air-pollution" argument was expected to be evaluated more positively than the "crash-risk" message (H1). The "air-pollution" and "crash-risk and air-pollution" messages were expected to be more effective than the "crash-risk" message on the behavioral intention to observe speed limits (H2a) and on the perceived efficacy of speed-limit observance in reducing air-pollution (H2b; Studies 1 and 2). Furthermore, positive framing was expected to be more effective than negative framing (H3), and presenting a message to be more effective than presenting no message (H4; Study 2). Broadly, our results argue in favor of our hypotheses. However in Study 2, the effects of message framing did not allow us to conclude that negative or positive framing was superior. All in all, messages with the "air-pollution" argument were more effective at leading drivers to observe speed limits. Thus, environmental protection may be a fruitful route to explore for increasing road safety.

Automated enumeration of CD34+ cells in peripheral blood and bone marrow.

We have developed a rapid and accurate method to enumerate the number of CD34+ cells in peripheral blood, bone marrow, and leukopheresis samples. The method consists of a two-tube assay and a dedicated software program for data acquisition and analysis. The first reagent combination consists of (a) a nucleic acid dye to identify nucleated cells, (b) a CD45 monoclonal antibody labeled with PE/CY5 to discriminate progenitor cells from mature lymphoid, neutrophil, erythroid, and monocytic cells, (c) an IgG1 control antibody labeled with PE to establish the boundary between specific and nonspecific staining, and (d) a known number of fluorescent beads to determine an absolute count of cells. In the second reagent combination the IgG1 control antibody is replaced by a CD34 antibody labeled with PE that is used to identify the CD34+ cells in the location established by the control reagent combination. The software program uses the fluorescent beads to adjust the forward light scatter, orthogonal light scatter, and three fluorescence detectors of the flow cytometer. The expected location of the CD34+ cells is then established with the control reagent combination followed by the enumeration of the CD34+ cells per microliter of sample with the reagent combination containing the CD34 antibody. This method is sensitive enough to detect CD34+ cells in peripheral blood of normal donors and can reliably determine an increase in CD34+ cells in the peripheral blood of patients treated with chemotherapy and/or growth factors. The method alleviates some of the difficulties encountered when small numbers of CD34+ cells are enumerated. The system allows for more precise evaluations of the grafts used for bone marrow transplantation.

Variations in the content of CD34+ cells in the peripheral blood of cancer patients receiving out-patient chemotherapy.

BACKGROUND: The routine measurement of CD34+ cells in non-mobilized peripheral blood, using flow cytometry, has been limited by the technical difficulty of measuring the absolute numbers of rare populations of cells. METHODS: We studied the numbers of CD34+ cells in the peripheral blood of 55 normal volunteers and 476 cancer patients receiving chemotherapy, in a university out-patient hematology/oncology clinic. Blood samples were stained with MAbs to CD34, CD45 and a DNA-specific dye, mixed with a defined number of fluorescent True-Count beads and analyzed by flow cytometry, using an automated acquisition and analysis program. RESULTS: The mean (+/- SD) CD34+ cell count among normal volunteers and previously untreated cancer patients were 1.4 +/- 1.4 and 1.8 +/- 2.8 CD34+ cells/microL, was stable among normal volunteers and patients receiving chemotherapy for prostate or breast cancer. In contrast, the CD34+ cell count among patients receiving dose-intensive weekly chemotherapy for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma varied between 1 cell/microL to > 300 CD34+ cells/microL in a predictable and cyclic fashion. DISCUSSION: The study demonstrates that the routine measurement of the number of CD34+ cells in peripheral blood samples can be performed using an automated single platform program in an out-patient setting. The availability of an automated data acquisition and analysis program could facilitate standardization of counting CD34+ cells in clinical samples analyzed at different laboratory sites.

Pathogenetic implications of internuclear bridging in myelodysplastic syndrome. An Eastern Cooperative Oncology Group/Southwest Oncology Group Cooperative Study.

Numerous morphologic features have been described in bone marrow and peripheral blood in myelodysplastic syndrome (MDS). We draw attention to a high incidence of a subtle morphologic feature, internuclear bridging (INB), not previously recognized as a morphologic feature in MDS. The occurrence of INB in MDS suggests an underlying abnormality of mitotic division that could explain the impaired production of hematopoietic cells, the addition and deletion cytogenetic changes, and the stepwise disease progression and cytogenetic progression characteristic of MDS. Lack of awareness that INB occurs in MDS may cause confusion of MDS and congenital dyserythropoietic anemia type I, a congenital process also characterized by INB.