Influence of the Extraction Process on the Characteristics of Romanian Mountain Walnut Oil

The objective of this work is to extract walnut oil using various processes in order to compare the influence on the nature of the components extracted, and thus identify the areas of potential use. We carried out the extractions by mechanical process, thanks to a press in reduced model provided with a worm. We obtained cold extracted oil whose characteristics slightly diverge from extra virgin oil found in shops in Romania, but its composition is similar. We were also able to extract by chemical process using two methods, Folch and Soxhlet. Commercially available table walnut oils are only cold extracted to avoid the presence of solvents. Those are difficult to remove and strongly oxidize the oil. Currently, consumers appreciate walnut oil for its taste and nutritional qualities. In nutrition, this oil is put forward for its composition rich in polyunsaturated fatty acids, which are needed for human body. Food supplements made from walnut oil are available today. For the moment, this is the only use of walnut oil. Indeed, there are some studies on other fields of application, but they remain in the field of research and nothing has yet been commercialized. In this present study, we compared the chemical and physical properties of cold-extracted oil with the solvent extraction of walnut kernel originating from the mountain region of Rumania. The cold extracted oil has a high content of polyunsaturated fatty acids (63%) and monounsaturated fatty acids (30%), a very low level of saturated fatty acid (7%) and no content of linolenic acid. The Soxhlet and Folch methods produced slightly different oils with increased amounts of minor components, which changes their characteristic. Even when solvent-extracted oils do not meet the standard criteria imposed by the Codex Alimentarius, they offer a possible use in the fields of food, cosmetics industries and biomedicine.

Cosmetic or Dietary Vegetable Oils Sampled in the Cameroonian Market May Not Expose Consumers to Lipid Oxidation Products Generating Oxidative Stress and Inflammation

Vegetable oils are a source of energy, essential fatty acids, antioxidants and fat-soluble vitamins useful for human health care and development. These oils also contribute to organoleptic quality of their products’ derivatives. However, their chemical and physical properties can be modified by the mode of their extraction, storage and distribution. These modifications might negatively affect the nutritional quality of the oils. The goals of this study were to: sample different vegetable oils for cosmetic or dietary use marketed in Cameroon, and verify purity and oxidation states of each kind of oil through determination of its acidity, iodine, peroxide, saponification, refractive indexes and the conformity of the labeling. The carotene content, the level of polar components and specific absorbance were also determined. As the result, six oils namely palm, palm kernel, coconut, black cumin, peanut and shea butter were collected. Apart from labeling, chemicals and physicals parameters analyzed were generally in accordance with the Cameroonian and Codex Alimentarius standard. This study suggests that vegetable oils sampled in the Cameroonian market may not expose consumers to lipid oxidation products generating pathological oxidative stress and inflammation. However, efforts in application of existing standard need to be done as far as labeling are concerned.

Micropropagation of the Moroccan Endemic Plant <i>Thymus broussonetii</i>Boiss. with Aromatic-Medicinal Value and Conservation Concern

Micropropagation from shoot tips and nodal segments was carried out for the conservation and domestication of spontaneous Moroccan thyme, Thymus broussonetii Boiss. subsp. broussonetii (endemic threatened). The mineral composition of the culture medium, as well as the succession of different growth regulators, influenced the in vitro growth of this species. Sterilized achenes of T. broussonetii were able to germinate on an agar medium containing Gautheret macronutrients with a rate of 25% and a degree of contamination of less than 4%. Shoot apices of 15-day seedlings (two cotyledon leaves) were cultivated on SD + 0.46 μM Kin medium and the explants obtained were transplanted every month. Six macronutrients (MS, B5, SH, SD, MSm and N30K) were tested and N30K was chosen for the following experiments. Seven cytokinins (Kin, BAP, 2iP, DPU, adenine, Zeat and TDZ) at 0.46, 0.93 and 2.32 μM/l were evaluated and the addition of 0.93 μM adenine to N30K medium favored significantly the induction of buds and the elongation of explants. Three polyamines (putrescine, spermidine and spermine) at 2, 5, 10 and 20 μM/l were tested. A better multiplication of buds, shoots and roots was noted for N30K + 10 μM spermine. Cytokinin-auxin combinations led to better root multiplication and an increase in the number of buds and the length of explants, particularly for 0.46 μM Kin + 2.85 μM IAA. Acclimatization was successfully carried out using vitroplants developing a good root system. One month after the start of acclimatization, 97% of T. broussonetii plantlets were healthy. Three months later, they were transplanted into larger pots. 100% of the acclimatized plants developed flowers in the 2nd year between June and August. Re-initiation of the in vitro culture was carried out from sterilized twig segments collected from the acclimatized plants of T. broussonetii with 1 - 2 nodes on the medium N30K + 0.46 μM Kin, and 52.1% of the explants healthily proliferated. Finally, two micropropagation prototypes were developed: shoot tip culture from seedlings obtained after germination of achenes and node culture from acclimatized plants.

Micropropagation and Acclimatization of Common Oregano (Origanum vulgare L. Subsp. vulgare) by Shoot Tip Culture

Origanum vulgare L. is a commercially valued species with remarkable biological properties. It is subject to over-exploitation practices that seriously threaten its sustainability for future generations. Thus, micropropagation serves as a tool for the protection and domestication of this species. In this study, we established an in vitro vegetative propagation protocol for Origanum vulgare. This is done through the axillary bud technique by carrying out various tests. Six culture media (MS, MSm, N30K, SD, SH and B5) were tested. Therefore, SD was chosen for the following experiments. Seven cytokinins (adenine (Ad), N6-(2-isopentenyl) (2ip), zeatin (Zeat), kinetin (Kin), benzyladenine (BAP), 1,3-diphenylurea (DPU) and thidiazuron (TDZ) at 5 concentrations (0.44, 1.33, 2.22, 3.11 and 4.44 μM/L) were evaluated. Thus, Kin at 3.11 μM allowed high regeneration of vitroplants, optimal elongation, total rooting of explants, maximum bud multiplication, and absence of hyperhydric explants. In fact, the integration of auxins (indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthaleneacetic acid (NAA)) into the culture medium and their combinations with 3.11 μM Kinetin contributed to the optimization of the root part. Thus, it was improved in particular in the case of 3.11 μM Kin and 6.27 μM IBA. Three polyamines (Putrescine, Spermidine and Spermine) at different concentrations (1.134, 3.402, 5.67, 7.938 and 11.34 μM/L) combined at 3.11 μM Kin and 6.27 μM IBA were tested. In fact, 1.304 μM putrescine was considered to be the most suitable for in vitro culture of explants, since it allowed optimal propagation of buds and roots, also a high rate of regeneration and rhizogenesis. GA3 at 1.15 μM combined with 3.11 μM Kin and 6.27 μM IBA permitted maximum bud multiplication. The acclimatization was carried out successfully using vitroplants showing good foliar and root development. Thus, three months after acclimatization, the seedlings were transferred into large pots under natural light and temperature conditions. Almost all acclimatized plants developed flowers in the first year between May and July.

Efficient Plant Regeneration from Explants of Compact Oregano (Origanum compactum Bentham)

Origanum compactum Benth., a species endemic to Morocco characterized by its biological activities, is overexploited because of its commercial value and threatened with extinction. Accordingly, measures for its conservation are needed. Micropropagation serves as a solution for the protection and the domestication of this species. In this investigation, we established a protocol for vegetative multiplication in vitro of Origanum compactum by the axillary bud technique. Six culture media (SH, SD, N30K, MS, MSm, B5) were tested to determine the most suitable mineral medium for growth and development of explants. Four cytokinins: Kinetin, Zeatin, BAP (6-Benzylaminopurine), 2ip (2-Isopentenyladenine) and three compounds with cytokinin activity: Adénine, 1,3-Diphenylurea (DPU) and Thidiazuron (TDZ) at five concentrations (0.44, 1.33, 2.22, 3.11, 4.44 μM) were tested on budding, growth, hyperhydria and rooting. Then three auxins, Indole-3-acetic acid (IAA), Naphthalene acetic acid (NAA) and Indole butyric acid (IBA) at four concentrations (1.14, 2.85, 4.56, 6.27 μM) in presence of 2.22 μM BAP were evaluated. The combination of Gibberellic acid (0.29, 1.5, 2.60, 2.89 μM GA3) and three polyamines (Putrescine, Spermidine, Spermine) at five concentrations (1.134, 3.402, 5.67, 7.938, 11.34 μM) with cytokinins and auxins were considered. Our results show that Margara medium is the most efficient and BAP at 2.22 μM is the best cytokinin for development of the aerial parts, with a regeneration rate of 88, 90%;rhizogenesis is successful with the combination of 6.27 μM IAA and 2.22 μM BAP. Moreover, the integration of 2.89 μM GA3 with 2.22 μM BAP and 6.27 μM IAA promotes vitroplant growth, bud and shoot multiplication and elongation of the aerial part. The addition of polyamines with 2.22 μM BAP and 6.27 μM IAA does not improve the root part, but Spermine at 5.67 μM promotes bud and shoot multiplication with a high percentage of regeneration, and spermidine at the same concentration gives long explants. Finally, plantlets with good root development were successfully acclimatized to natural conditions and served as a source to establish in vitro culture again.

Interaction Effect of Plant Growth Regulators on Shoot Micropropagation of Aromatic Plant Origanum elongatum (Bonnet) Emberger &Maire

Origanum elongatum (Bonnet) Emb. & Maire, is a medicinal, aromatic and endemic plant of Morocco, characterized by its pharmacological effects, and is commonly used for the production of essential oils and aromas, resulting in high harvest and overexploitation pressure. This is why the present study aims to implement the in vitro micropropagation of Origanum elongatum for optimal vitroplant production. Six macroelements were tested (SH, SD, N30K, MS, MSm and B5) and the SD medium was selected for vegetative propagation of the explants. Seven cytokinins: adenine (Ad), N6-(2-Isopentenyl) adenine, zeatin (Zeat), kinetin (Kin), benzyladenine (BAP), 1,3-diphenylurea (DPU) and thidiazuron (TDZ) were then evaluated at five concentrations (0.44, 1.33, 2.22, 3.11 and 4.44 μM/L) on growth, development, budding, rooting and hyperhydricity. 0.44 μM Kin was selected and combined with three auxins: indole-3-acetic acid (IAA), indole-3-butyric acid (AIB), and 1-naphthaleneacetic acid (NAA) at four concentrations (1.14, 2.85, 4.56 and 6.27 μM/L) to improve rooting and association with 1.14 μM IAA was shown to be efficient for roots development. Different concentrations of gibberellic acid (0.29, 1.5, 2.60 and 2.89 μM/L), combined with 0.44 μM/L Kin and 1.14 μM/L IAA, were tested and 2.60 μM/L GA3 gave maximum buds and shoots. Then, the combination of three polyamines at five concentrations (1.134, 3.402, 5.67, 7.938 and 11.34 μM/L) with 0.44 μM Kin and 1.14 μM/L IAA showed an increase in the number of buds and shoots for 7.938 μM/L putrescine and 3.402 μM/L spermine. Finally, seedlings with good foliar and root development were acclimatized.

Biology and Physiology of <I>Colletotrichum acutatum</I>Strains Causing Strawberry’s Anthracnose

Seven Colletotrichum acutatum strains isolated from strawberries were cultivated on various culture media and tested in vitro and in vivo on Fragaria x ananassa for anthracnose symptoms. PDA caused an optimum growth of all isolates, MEA, ML and Strawberry allowed a good but not optimal growth. Czapeck, Sabouraud and the organic medium from potato gave the lowest growth rate of all isolates. PDA allowed a good sporulation of isolates follow-up by strawberry, MEA;in contrast, ML, Czapeck and Sabouraud gave a low sporulation. The fungal development is maximal at 25&degC and 27&degC for all the isolates studied. No growth was observed at 5&degC and 37&degC. The studied strains developed at all pH values. They didn’t develop at 0.6, 0.65 and 0.7 aw but mycelial growth was perfect at 1 and 0.95 aw. The in vitro test of the pathogenicity caused by C. acutatum strains on strawberry’s leaves showed an increasing percentage of infection with time and different infection rates among C. acutatum strains, strain Ca6 having a pathogenic power very high compared to the other isolates. After in vivo inoculation and incubation of the seedlings, all isolates caused severe symptoms related to anthracnose on leaflets and petioles of the studied strawberry plants.

Effect of Rosehip (<i>Rosa Canina</i>) Extracts on Human Brain Tumor Cell Proliferation and Apoptosis

Rosehips are blossoms from the wild rose (Rosa canina) and are commonly used as an herbal remedy. Previous reports have shown that extracts made from rosehip plants are able to reduce cell proliferation of cancer cells. In this study, we investigated the efficacy of rosehip extracts in preventing cell proliferation of three human glioblastoma cell lines A-172, U-251 MG and U-1242 MG cell lines. Each of the glioblastoma cell lines treated with rosehip extracts (1 mg/mL-25 ng/mL) demonstrated a significant decrease in cell proliferation. The rosehip extract-mediated decrease in cell proliferation was equal to or better than the decrease of cell proliferation observed when inhibitors of the MAPK (U0126, 10 μM) or AKT (LY294002, 20 μM) signaling pathways were utilized. Additionally, pretreatment of the these cell lines with Rosehip extracts (1 mg/mL-25 ng/mL) selectively decreased AKT, MAPK, and p70S6K phosphorylation suggesting these extracts prevent glioblastoma multiforme cell proliferation by blocking both the MAPK and AKT signaling mechanisms. Results from colorimetric cell death assays, cell cycle analysis by flow cytometry, as well as western blot studies demonstrate that rosehip extracts inhibit cell proliferation but do not promote apoptosis. Moreover, rosehip extracts were able to increase the efficacy of Temozolomide, a chemotherapeutic agent used to treat patients with glioblastomas. Surprisingly, rosehip extracts demonstrated a greater inhibition of cell proliferation than in combination with Temozolomide (100 μM) or Temozolomide as a single agent. Taken together these data suggest that rosehip extracts are capable of decreasing glioblastoma cell proliferation without promoting apoptosis and demonstrate a greater cell proliferation inhibitory effect than Temozolomide. More importantly, rosehip extracts may serve as an alternative or compliment to current chemotherapeutic regimens for glioblastomas.

Evaluation of Plant Growth Promoting Ability of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum TR In Vivo

Bacillus amyloliquefaciens Bc2 and Trichderma harzianum TR were used to evaluate their growth-promoting activity on cultivated strawberries, under laboratory and field conditions, and we have noticed that the percentage of achene germination is important for ones treated with TR (=97%) followed by those treated with Bc2 strain (=90%) and the control (=84%). Inoculations on field showed that on untreated soil with insecticide, TR is effective and allows the development of plants and extends the duration of flowering and fruiting. On treated soil, Bc2 clearly promotes the growth and development of strawberry seedlings and its role as plant growth promoting microorganisms has been proved.

Biocontrol Potential of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum TR against Strawberry Anthracnose under Laboratory and Field Conditions

The increasingly strict regulation of the use of phytosanitary products and the will of the populations to move towards sustainable development allow bio-sourced products and more particularly for biocontrol in the field of pesticides to progress. The study carried out here concerns the evaluation of the potency of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum to fight against strawberry cultures infected with anthracnose (Fragaria × ananassa Duch.). The studies were carried out in the laboratory and in the field. The results indicate that B. amyloliquefaciens Bc2 and Trichoderma harzianum TR are effective for the biological control of anthracnose, gray mold and powdery mildew on strawberries grown in field conditions. The increase in plant size and the number of fruits produced with these control agents has also been observed.

In Vitro Antagonistic Activity of Trichoderma harzianum and Bacillus amyloliquefaciens against Colletotrichum acutatum

Biological control of plant diseases makes extensive use of the antagonists. The effectiveness of the control depends on the choice of an effective antagonist strains from criteria of implying a good knowledge of biological peculiarities of the material used. In this study, the antagonistic activities of nine isolates of Bacillus amyloliquefaciens and one isolate of Trichoderma harzianum were tested in vitro against seven strains of Colletotrichum acutatum, the causal agent of strawberry anthracnose. It was found that T. harzianum and B. amyloliquefaciens strains showed the ability to inhibit the mycelial growth of the pathogen by dual culture technique by more than 50 percent. The nonvolatile substances produced by the antagonists showed high inhibition percentages that are more than 99 percent, meanwhile their volatile compounds inhibited the mycelial growth of C. acutatum strains with low inhibition percentages starting from 30 percent except the substances produced by B. amyloliquefaciens Bc2 that showed a high inhibition percentage of more than 70 percent.

Effect of Amino Acids on Secondary Somatic Embryogenesis of Moroccan Cork Oak (<i>Quercus suber</i>L.) Tree

In the present study, we tested the effect of amino acids on secondary somatic embryogenesis of Moroccan cork oak (Quercus suber L.). Secondary mature and immature somatic embryos were obtained from primary somatic embryos cultured in N30K medium supplemented with nineteen amino acids. Stimulation of embryogenesis was dependent on the type and concentration of amino acid in the medium. Thus, L-glutamine and L-asparagine at 3.42 mM have been proved to be the most favorable for the formation of functional somatic embryos and the induction of secondary somatic embryos.

Micropropagation of Carob(Ceratonia siliqua L.)through Adventitious Buds of Immature Embryonic Cotyledons

Adventitious budding from embryonic cotyledons of immature seeds of carob was obtained. The combination of BAP (4.44 μM) and NAA (1.5 μM) furthered the neoformation of adventitious buds. These latter were multiplied on MS medium added with BAP (2.22 μM). Stems and leaves growing were improved by adding 2.02 μM GA3. Elongation was favored by 0.5 μM NAA. 70% of rooting was obtained with 10 μM IBA.

Addressing the Security Challenges of Big Data Analytics in Healthcare Research

Big data and associated analytics have the potential to revolutionize healthcare through the tools and techniques they offer to manage and exploit the large volumes of heterogeneous data being collected in the healthcare domain. The strict security and privacy constraints on this data, however, pose a major obstacle to the successful use of these tools and techniques. The paper first describes the security challenges associated with big data analytics in healthcare research from a unique perspective based on the big data analytics pipeline. The paper then examines the use of data safe havens as an approach to addressing the security challenges and argues for the approach by providing a detailed introduction to the security mechanisms implemented in a novel data safe haven. The CIMVHR Data Safe Haven (CDSH) was developed to support research into the health and well-being of Canadian military, Veterans, and their families. The CDSH is shown to overcome the security challenges presented in the different stages of the big data analytics pipeline.

Effect of Different Concentrations of the Suspensions of B.amyloliquefaciens and T.harzianum on the Development of C.acutatum

The effect of nine isolates of Bacillus amyloliquefaciensand one strain of Trichoderma harzianum, TR, on mycelial growth and germination of Colletotrichum acutatum were studied. The nine isolates were identified as Bacillus amyloliquefaciens. The efficacy of isolates was tested, at different concentrations. Results showed that one Bacillus isolates (Bc2) and TR were more effective at the lower concentration tested (3 × 105 CFU/ml and 105 conidia/ml).

High-efficiencygenomeediting in plants mediated by a Cas9 gene containing multiple introns

The recent discovery of the mode of action of the CRISPR/Cas9 systemhas provided biologists with a useful tool for generating site-specific mutations in genes of interest.In plants,site-targeted mutations are usually obtained by the stable transformation of a Cas9 expression construct into the plant genome.The efficiency of introducing mutations in genes of interest can vary considerably depending on the specific features of the constructs,including the source and nature of the promoters and terminators used for the expression of the Cas9 gene and the guide RNA,and the sequence of the Cas9 nuclease itself.To optimize the efficiency of the Cas9 nuclease in generating mutations in target genes in Arabidopsis thaliana,we investigated several features of its nucleotide and/or amino acid sequence,including the codon usage,the number of nuclear localization signals(NLSs),and the presence or absence of introns.We found that the Cas9 gene codon usage had some effect on its activity and that two NLSs worked better than one.However,the highest efficiency of the constructs was achieved by the addition of 13 introns into the Cas9 coding sequence,which dramatically improved the editing efficiency of the constructs.None of the primary transformants obtained with a Cas9 gene lacking introns displayed a knockout mutant phenotype,whereas between 70%and 100%of the primary transformants generated with the intronized Cas9 gene displayed mutant phenotypes.The intronized Cas9 gene was also found to be effective in other plants such as Nicotiana benthamiana and Catharanthus roseus.