scholarly journals Rapid Rubber Extraction and NMR Spectroscopy of Rubber, Extracted from the Endemic Species Scorzonera Tau-Saghyz

2020 ◽  
Vol 22 (1) ◽  
pp. 59
Author(s):  
K.K. Boguspaev ◽  
S.K. Turasheva ◽  
T.M. Seilkhanov ◽  
D.G. Faleev ◽  
M.S. Mutalkhanov ◽  
...  
Keyword(s):  
Adventitious Shoots ◽  
Physiological Age ◽  
Root Extract ◽  
Nmr Characterization ◽  
First Time ◽  
Rubber Good ◽  
Isoprene Rubber

Scorzonera tau-saghyz Lipsch. et G.G. Bosse is an endemic rubber producing plant, growing in mountain regions in South Kazakhstan. The rubber content in plants and the quality of biopolymer has an important impact on industrial rubber production. The results of this study showed that the amount of rubber in S. tau-saghyz roots fluctuates between 7.74% and 38.75%. The amount of synthesized and deposited rubber biopolymer particles depends on various factors such as physiological age of plant, origin, temperature, moisture and environmental conditions. We optimized the extraction method of natural rubber by using n-hexane as a solvent for direct extraction. This method allows extracting the maximum amount of rubber from 3‒4-year-old plants. NMR results show structural links of natural isoprene rubber in the root extract sample. There is a clear relationship between methyl, methine and methylene protons which corresponds to isoprene rubber structure. The samples having strongly marked singlets that are inherent for rubber functional groups confirms the stereospecific structure of rubber. Good solubility of the root extract in deuterated chloroform can characterize the low molecular weight of the polymer. NMR characterization of rubber, extracted from S. tau-saghyz roots, is reported for the first time. Regeneration in vitro provides an important opportunity for endemic preservation by rapidly increasing the number of plants. The best regeneration of adventitious shoots was obtained on MS medium containing 5.5 μM kinetin and 0.5 μM NAA. The plants were successfully acclimatized in a glasshouse with 75% of S. tau-saghyz plantlets, respectively surviving after transfer to ex vitro conditions.

scholarly journals PPIP5K2 and PCSK1 are Candidate Genetic Contributors to Familial Keratoconus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mariam Lofty Khaled ◽  
Yelena Bykhovskaya ◽  
Chunfang Gu ◽  
Alice Liu ◽  
Michelle D. Drewry ◽  
...  
Keyword(s):  
Gene Trap ◽  
Whole Exome ◽  
The Us ◽  
Related Quality ◽  
First Time

AbstractKeratoconus (KC) is the most common corneal ectatic disorder affecting >300,000 people in the US. KC normally has its onset in adolescence, progressively worsening through the third to fourth decades of life. KC patients report significant impaired vision-related quality of life. Genetic factors play an important role in KC pathogenesis. To identify novel genes in familial KC patients, we performed whole exome and genome sequencing in a four-generation family. We identified potential variants in the PPIP5K2 and PCSK1 genes. Using in vitro cellular model and in vivo gene-trap mouse model, we found critical evidence to support the role of PPIP5K2 in normal corneal function and KC pathogenesis. The gene-trap mouse showed irregular corneal surfaces and pathological corneal thinning resembling KC. For the first time, we have integrated corneal tomography and pachymetry mapping into characterization of mouse corneal phenotypes which could be widely implemented in basic and translational research for KC diagnosis and therapy in the future.

scholarly journals In vitro cytotoxicity of Aspilia pluriseta Schweinf. extract fractions

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Sospeter N. Njeru ◽  
Jackson M. Muema
Keyword(s):  
Biological Activities ◽  
Vero Cells ◽  
In Vitro Cytotoxicity ◽  
Root Extract ◽  
Lack Of Information ◽  
Information Gap ◽  
Diphenyltetrazolium Bromide ◽  
Potential Use

Abstract Objectives We and others have shown that Aspilia pluriseta is associated with various biological activities. However, there is a lack of information on its cytotoxicity. This has created an information gap about the safety of A. pluriseta extracts. As an extension to our recent publication on the antimicrobial activity and the phytochemical characterization of A. pluriseta root extracts, here we report on cytotoxicity of tested solvent fractions. We evaluated the potential cytotoxicity of these root extract fractions on Vero cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results We show that all solvent extract fractions (except methanolic solvent fractions) had cytotoxic concentration values that killed 50% of the Vero cells (CC50) greater than 20 µg/mL and selectivity index (SI) greater than 1.0. Taken together, we demonstrate that, A. pluriseta extract fractions’ earlier reported bioactivities are within the acceptable cytotoxicity and selective index limits. This finding scientifically validates the potential use of A. pluriseta in the discovery of safe therapeutics agents.

scholarly journals Characterization of Postharvest Fungicide-Resistant Botrytis cinerea Isolates From Commercially Stored Apple Fruit

2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Apple Fruit ◽  
Sublethal Dose ◽  
Long Term Storage ◽  
Pose Control ◽  
First Time

Botrytis cinerea causes gray mold and is an economically important postharvest pathogen of fruit, vegetables, and ornamentals. Fludioxonil-sensitive B. cinerea isolates were collected in 2011 and 2013 from commercial storage in Pennsylvania. Eight isolates had values for effective concentrations for inhibiting 50% of mycelial growth of 0.0004 to 0.0038 μg/ml for fludioxonil and were dual resistant to pyrimethanil and thiabendazole. Resistance was generated in vitro, following exposure to a sublethal dose of fludioxonil, in seven of eight dual-resistant B. cinerea isolates. Three vigorously growing B. cinerea isolates with multiresistance to postharvest fungicides were further characterized and found to be osmosensitive and retained resistance in the absence of selection pressure. A representative multiresistant B. cinerea strain caused decay on apple fruit treated with postharvest fungicides, which confirmed the in vitro results. The R632I mutation in the Mrr1 gene, associated with fludioxonil resistance in B. cinerea, was not detected in multipostharvest fungicide-resistant B. cinerea isolates, suggesting that the fungus may be using additional mechanisms to mediate resistance. Results from this study show for the first time that B. cinerea with dual resistance to pyrimethanil and thiabendazole can also rapidly develop resistance to fludioxonil, which may pose control challenges in the packinghouse environment and during long-term storage.

scholarly journals Identification and characterization of molecular interactions between mortalin/mtHsp70 and HSP60

2005 ◽  
Vol 391 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Renu Wadhwa ◽  
Syuichi Takano ◽  
Kamaljit Kaur ◽  
Satoshi Aida ◽  
Tomoko Yaguchi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Molecular Interactions ◽  
Heat Shock Protein 60 ◽  
Hsp60 Expression ◽  
Mitochondrial Hsp70 ◽  
Shock Proteins ◽  
First Time

Mortalin/mtHsp70 (mitochondrial Hsp70) and HSP60 (heat-shock protein 60) are heat-shock proteins that reside in multiple subcellular compartments, with mitochondria being the predominant one. In the present study, we demonstrate that the two proteins interact both in vivo and in vitro, and that the N-terminal region of mortalin is involved in these interactions. Suppression of HSP60 expression by shRNA (short hairpin RNA) plasmids caused the growth arrest of cancer cells similar to that obtained by suppression of mortalin expression by ribozymes. An overexpression of mortalin, but not of HSP60, extended the in vitro lifespan of normal fibroblasts (TIG-1). Taken together, this study for the first time delineates: (i) molecular interactions of HSP60 with mortalin; (ii) their co- and exclusive localizations in vivo; (iii) their involvement in tumorigenesis; and (iv) their functional distinction in pathways involved in senescence.

scholarly journals THE PRODUCTION OF MONOCLONAL ANTIBODIES TO TETRASACCHARIDE - SYNTHETIC ANALOGUE OF THE CAPSULAR POLYSACCHARIDE OF STREPTOCOCCUS PNEUMONIAE OF SEROTYPE 14 AND THEIR IMMUNOCHEMICAL CHARACTERIZATION

2018 ◽  
pp. 26-31
Author(s):  
I. V. Yakovleva ◽  
E. A. Kurbatova ◽  
E. A. Akhmatova ◽  
E. V. Sukhova ◽  
D. V. Yashunsky ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Streptococcus Pneumoniae ◽  
Capsular Polysaccharide ◽  
Solid Phase ◽  
Synthetic Analogue ◽  
Immunochemical Characterization ◽  
Carbohydrate Epitopes ◽  
First Time

Aim. Production of monoclonal antibodies (mAb) to synthetic tetrasaccharide - repeating unit of the capsular polysaccharide (CP) of Streptococcus pneumoniae serotype 14 and their immunochemical characterization. Materials and methods. In order to generate the hybridoma producing mAb, mice were immunized with synthetic tetrasaccharide conjugated with bovine serum albumin (BSA) with following hybridization of B lymphocytes with mouse myeloma cells. Antibodies were obtained in vitro andin vivo. Immunochemical characterization of mAb to tetrasaccharide was carried out using a variety of ELISA options. Results. For the first time obtained mouse hybridoma, producing IgM to tetrasacchride. The IgM titer of anti-tetrasacharide antibodies in supernatants of clones and in the ascitic fluid of mice in ELISA detected by biotinylated tetrasaccharide and synthetic CP adsorbed on the solid phase was higher compared to the use of bacterial CP as well cover antigen. In the reaction of inhibition of the ELISA, the mAb recognized the corresponding carbohydrate epitopes of the bacterial CP of S. pneumoniae serotype 14 dissolved in the liquid phase better than tetrasaccharide ligand and synthetic CP. Conclusion. To detect mAb to tetrasaccharide in ELISA preferably to use synthetic analogues of the CP as solid phase antigens. The obtained mAb to tetrasaccharide can be used to determine the representation of the protective tetrasaccharide epitope of CP in the development of pneumococcal vaccines.

scholarly journals Immunological changes in growing mice fed on diets containing casein or peas (Pisum sativum var. Belinda) as the source of protein

1995 ◽  
Vol 73 (1) ◽  
pp. 87-97 ◽  
Author(s):  
J.Alfredo Martínez ◽  
M. Luisa Esparza ◽  
Jesús Larralde
Keyword(s):  
Pisum Sativum ◽  
Individual Performance ◽  
Gastrointestinal Disorders ◽  
Antigenic Proteins ◽  
Poor Individual ◽  
First Time ◽  
Different Sources

The effects of two different sources of protein: peas (Pisum sativum var. Belinda) and casein on immunocompetence, nutritional utilization and growth performance have been investigated in recently weaned mice. Feeding these animals on a pea diet resulted in an impairment in growth and significant decreases in the weights of liver, muscle, kidneys and femur, while intestine weights increased. No differences in food consumption were observed, but food conversion efficiency (food intake: weight gain) was increased in pea-fed animals compared with those offered the casein diet. Packed cell volume and serum Fe and Zn levels fell significantly after legume-protein intake, and, by contrast, Cu values increased slightly. Serum albumin levels showed a statistically significant reduction in mice fed on the diet containing peas. However, y-globulins and immunoglobulin G titres were markedly increased. The characterization of spleen-cell subsets using monoclonal antibodies revealed a significantly higher percentage of T-lymphocytes in the pea group compared with casein-fed animals, while no changes were observed in the proportions of B-lymphocytes and macrophages. In vitro mitogenic responses to phytohaemagglutinin, concanavalin A and Escherichia coli lipopolysaccharide S were slightly, but not significantly, lower in the pea-fed animals. Our results describe, apparently for the first time in mice, some immunological disturbances after peak intake. These results may lead to a better understanding of the possible role of antigenic proteins in gastrointestinal disorders and the poor individual performance after legume intake.

scholarly journals Characterization of quality of sediments from Paranaguá Bay (Brazil) by combined in vitro bioassays and chemical analyses

2016 ◽  
Vol 36 (7) ◽  
pp. 1811-1819 ◽  
Author(s):  
Juliane Rizzi ◽  
Elisabet Pérez-Albaladejo ◽  
Denise Fernandes ◽  
Javier Contreras ◽  
Sandro Froehner ◽  
...  
Keyword(s):  
Chemical Analyses ◽  
Paranaguá Bay ◽  
In Vitro Bioassays

scholarly journals The Relaxase of the Rhizobium etli Symbiotic Plasmid Shows nic Site cis-Acting Preference

2006 ◽  
Vol 188 (21) ◽  
pp. 7488-7499 ◽  
Author(s):  
Daniel Pérez-Mendoza ◽  
María Lucas ◽  
Socorro Muñoz ◽  
José A. Herrera-Cervera ◽  
José Olivares ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Rhizobium Etli ◽  
Cis Acting ◽  
Cleavage Activity ◽  
Symbiotic Plasmid ◽  
The Family ◽  
Nick Site ◽  
First Time

ABSTRACT Genetic and biochemical characterization of TraA, the relaxase of symbiotic plasmid pRetCFN42d from Rhizobium etli, is described. After purifying the relaxase domain (N265TraA), we demonstrated nic binding and cleavage activity in vitro and thus characterized for the first time the nick site (nic) of a plasmid in the family Rhizobiaceae. We studied the range of N265TraA relaxase specificity in vitro by testing different oligonucleotides in binding and nicking assays. In addition, the ability of pRetCFN42d to mobilize different Rhizobiaceae plasmid origins of transfer (oriT) was examined. Data obtained with these approaches allowed us to establish functional and phylogenetic relationships between different plasmids of this family. Our results suggest novel characteristics of the R. etli pSym relaxase for previously described conjugative systems, with emphasis on the oriT cis-acting preference of this enzyme and its possible biological relevance.

Comparative in vitro and early nursery performance of adventitious shoots from cryopreserved cotyledons and axillary shoots from epicotyls of the same zygotic embryo of control-pollinated Pinus radiata

2005 ◽  
Vol 35 (11) ◽  
pp. 2629-2641 ◽  
Author(s):  
Cathy L Hargreaves ◽  
Lynette J Grace ◽  
Susan A van der Maas ◽  
Mike I Menzies ◽  
Satish Kumar ◽  
...  
Keyword(s):  
Pinus Radiata ◽  
Zygotic Embryo ◽  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Radiata Pine ◽  
Field Trials ◽  
Physiological Age ◽  
In Vitro Growth ◽  
Axillary Shoots

This is the first published report comparing production and performance of adventitious shoots from cryopreserved cotyledons, with axillary shoots formed from epicotyls of the same zygotic embryo of radiata pine (Pinus radiata D. Don). Genotypes from 10 control-pollinated families of P. radiata in two treatments were compared for shoot initiation, in vitro growth, rooting, and early nursery performance. Plant growth in nursery beds was assessed by measuring height after 2 and 7 months. After 8 months in nursery beds, the physiological ages of genotypes were assessed before field planting. Genotype capture was higher from the cryopreserved cotyledons than from the epicotyls. This technique has the advantage of preserving juvenile material while field testing is done. Early shoot multiplication in both treatments was good. After four transfer cycles, epicotyl cultures showed improved elongation and higher multiplication rates. After 6 months of in vitro growth, shoots from both treatments were given auxin pulses. Shoots of adventitious origin were slower to root than epicotyl-derived shoots. Overall rooting rates were satisfactory. Plants of adventitious origin were shorter when planted into nursery beds and when transferred to field trials. Assessment of relative physiological age indicated that all plants of adventitious origin showed some increase.

scholarly journals Characterization of the secretory profile and exosomes of limbal stem cells in the canine species

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0244327
Author(s):  
Antonio J. Villatoro ◽  
Cristina Alcoholado ◽  
María del Carmen Martín-Astorga ◽  
Gustavo Rico ◽  
Viviana Fernández ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ocular Surface ◽  
Inhibitory Effect ◽  
Proteomic Profile ◽  
Limbal Stem Cells ◽  
High Concentrations ◽  
First Time

Limbal stem cells (LSCs) are a quiescent cell population responsible for the renewal of the corneal epithelium. Their deficiency is responsible for the conjunctivization of the cornea that is seen in different ocular pathologies, both in humans and in the canine species. The canine species represents an interesting preclinical animal model in ocular surface pathologies. However, the role of LSCs in physiological and pathological conditions in canine species is not well understood. Our objective was to characterize for the first time the soluble factors and the proteomic profile of the secretome and exosomes of canine LSCs (cLSCs). In addition, given the important role that fibroblasts play in the repair of the ocular surface, we evaluated the influence of the secretome and exosomes of cLSCs on their proliferation in vitro. Our results demonstrated a secretory profile of cLSCs with high concentrations of MCP-1, IL-8, VEGF-A, and IL-10, as well as significant production of exosomes. Regarding the proteomic profile, 646 total proteins in the secretome and 356 in exosomes were involved in different biological processes. Functionally, the cLSC secretome showed an inhibitory effect on the proliferation of fibroblasts in vitro, which the exosomes did not. These results open the door to new studies on the possible use of the cLSC secretome or some of its components to treat certain pathologies of the ocular surface in canine species.

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