Characterization of Two BAHD Acetyltransferases Highly Expressed in the Flowers of Jasminum sambac (L.) Aiton

Volatile benzenoid compounds are found in diverse aromatic bouquets emitted by most moth-pollinated flowers. The night-blooming Jasminum sambac is widely cultivated worldwide in the tropics and subtropics for ornamental and industrial purposes owing to its fragrant flowers. Benzylacetate is a characteristic constituent in jasmine scent which makes up to approximately 20–30% of the total emission in the headspace or extract, but the biosynthesis enzymes and the encoding genes have not yet been described. Here, we identify two cytosolic BAHD acyltransferases specifically expressed in the petals with a positive correlation closely to the emission pattern of the volatile benzenoids. Both JsBEAT1 and JsBEAT2 could use benzylalcohol and acetate-CoA as substrates to make benzylacetate in vitro. The recombinant GST-JsBEAT1 has an estimated apparent Km of 447.3 μM for benzylalcohol and 546.0 μM for acetate-CoA, whereas in the instance of the His-JsBEAT2, the Km values are marginally lower, being 278.7 and 317.3 μM, respectively. However, the catalytic reactions by the GST-JsBEAT1 are more efficient than that by the His-JsBEAT2, based on the steady-state kcat parameters. Furthermore, ectopic expression of JsBEAT1 and JsBEAT2 in the transgenic P. hybrida plants, driven by a flower-specific promotor, significantly enhances the biosynthesis of benzylbenzoate and benzylacetate, as well as the total VOCs.

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Characterization of Two Ethephon-Induced IDA-Like Genes from Mango, and Elucidation of Their Involvement in Regulating Organ Abscission

Genes ◽

10.3390/genes12030439 ◽

2021 ◽

Vol 12 (3) ◽

pp. 439

Author(s):

Avinash Chandra Rai ◽

Eyal Halon ◽

Hanita Zemach ◽

Tali Zviran ◽

Isaac Sisai ◽

...

Keyword(s):

Expression Profiles ◽

Mangifera Indica ◽

Ectopic Expression ◽

Fruit Trees ◽

Floral Organ ◽

Cytosolic Ph ◽

Early Increase ◽

Encoding Genes ◽

Fruitlet Abscission

In mango (Mangifera indica L.), fruitlet abscission limits productivity. The INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) peptide acts as a key component controlling abscission events in Arabidopsis. IDA-like peptides may assume similar roles in fruit trees. In this study, we isolated two mango IDA-like encoding-genes, MiIDA1 and MiIDA2. We used mango fruitlet-bearing explants and fruitlet-bearing trees, in which fruitlets abscission was induced using ethephon. We monitored the expression profiles of the two MiIDA-like genes in control and treated fruitlet abscission zones (AZs). In both systems, qRT-PCR showed that, within 24 h, both MiIDA-like genes were induced by ethephon, and that changes in their expression profiles were associated with upregulation of different ethylene signaling-related and cell-wall modifying genes. Furthermore, ectopic expression of both genes in Arabidopsis promoted floral-organ abscission, and was accompanied by an early increase in the cytosolic pH of floral AZ cells—a phenomenon known to be linked with abscission, and by activation of cell separation in vestigial AZs. Finally, overexpression of both genes in an Atida mutant restored its abscission ability. Our results suggest roles for MiIDA1 and MiIDA2 in affecting mango fruitlet abscission. Based on our results, we propose new possible modes of action for IDA-like proteins in regulating organ abscission.

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Preparation and Characterization of Intramuscular PLGA Based Microsphere

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v9i5.3631 ◽

2019 ◽

Vol 9 (5) ◽

pp. 127-130

Author(s):

Prateek Pathak ◽

Sarvesh Paliwal

Keyword(s):

Steady State ◽

Oral Therapy ◽

Extended Release ◽

In Vitro Study ◽

In Vitro Dissolution ◽

Plga Microspheres ◽

Anti Psychotic ◽

Multiple Dosing

The current studies was design and evaluate biodegradable PLGA microspheres for sustained or extended release, with primary goal of avoiding combination of oral therapy for the treatment of schizophrenia. PLGA copolymers 75:25 was used to prepare four microsphere formulations of anti-psychotic drug Olanzapine. The microspheres were characterized by in-vitro dissolution and other physio-chemical methods. A simulation of multiple dosing at weekly or 15-day regimen revealed pulsatile behavior for all formulations with steady state may be achieved by the second dose. Overall, the in-vitro study of Formulations 001, 002, 003, or 004 will eliminate the need for combination oral therapy and reduce time to achieve steady state, with a smaller washout period upon duration of therapy. Results of this study prove the suitability of using PLGA 75:25 copolymers of different composition and molecular weight to produce sustained or extended release formulations that can enhance pharmacological effectiveness for anti-psychotic intra-muscular administration of Olanzapine. Keywords: sustained release, PLGA microspheres, Olanzapine

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Characterization of two catalase-peroxidase-encoding genes in Fusarium verticillioides reveals differential responses to in vitro versus in planta oxidative challenges

Molecular Plant Pathology ◽

10.1111/mpp.12591 ◽

2017 ◽

Vol 19 (5) ◽

pp. 1127-1139 ◽

Cited By ~ 7

Author(s):

Shan Gao ◽

Scott E. Gold ◽

Anthony E. Glenn

Keyword(s):

Fusarium Verticillioides ◽

In Planta ◽

Catalase Peroxidase ◽

Differential Responses ◽

Encoding Genes

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In vitro characterization of neural stem cells: Functional response to the enteric neurotrophin GDNF and co-culture with human colonic smooth muscle

Gastroenterology ◽

10.1016/s0016-5085(01)80307-3 ◽

2001 ◽

Vol 120 (5) ◽

pp. A62-A62

Author(s):

M MICCI ◽

R LEARISH ◽

P PASRICHA

Keyword(s):

Stem Cells ◽

Smooth Muscle ◽

Neural Stem Cells ◽

Functional Response ◽

In Vitro Characterization

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Oxidative stress as antifibrogenic stimulus? Low dose steady state H2O2 induces fibrolytic MMP–3 in vitro and in vivo

Zeitschrift für Gastroenterologie ◽

10.1055/s-0031-1295764 ◽

2012 ◽

Vol 50 (01) ◽

Author(s):

G Millonig ◽

S Ottinger ◽

HK Seitz ◽

S Mueller

Keyword(s):

Oxidative Stress ◽

Steady State ◽

Low Dose

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Biological and physico-chemical characterization of ultra high diluted Hydrastis canadensis and in vitro studies on mammalian cells

Allgemeine Homöopathische Zeitung ◽

10.1055/s-0037-1601241 ◽

2017 ◽

Vol 262 (02) ◽

pp. 2-76

Author(s):

N Chandrakar ◽

MK Temgire ◽

SG Kane ◽

AK Suresh ◽

J Bellare

Keyword(s):

Mammalian Cells ◽

Chemical Characterization ◽

In Vitro Studies ◽

Hydrastis Canadensis ◽

Physico Chemical

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Assays for Phospholipid-Dependent Formation of Thrombin and Xa: A Potential Method for Quantifying Lupus Anticoagulant Activity

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646437 ◽

1991 ◽

Vol 66 (04) ◽

pp. 453-458 ◽

Cited By ~ 15

Author(s):

John T Brandt

Keyword(s):

Specific Activity ◽

Anticoagulant Activity ◽

Factor Xa ◽

Clinical Importance ◽

Potential Method ◽

Quantitative Expression ◽

Increased Risk ◽

Apparent Association

SummaryLupus anticoagulants (LAs) are antibodies which interfere with phospholipid-dependent procoagulant reactions. Their clinical importance is due to their apparent association with an increased risk of thrombo-embolic disease. To date there have been few assays for quantifying the specific activity of these antibodies in vitro and this has hampered attempts to purify and characterize these antibodies. Methods for determining phospholipid-dependent generation of thrombin and factor Xa are described. Isolated IgG fractions from 7 of 9 patients with LAs were found to reproducibly inhibit enzyme generation in these assay systems, permitting quantitative expression of inhibitor activity. Different patterns of inhibitory activity, based on the relative inhibition of thrombin and factor Xa generation, were found, further substantiating the known heterogeneity of these antibodies. These systems may prove helpful in further purification and characterization of LAs.

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Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648381 ◽

1992 ◽

Vol 67 (01) ◽

pp. 063-065 ◽

Cited By ~ 6

Author(s):

Sherryl A M Taylor ◽

Jacalyn Duffin ◽

Cherie Cameron ◽

Jerome Teitel ◽

Bernadette Garvey ◽

...

Keyword(s):

Nucleotide Substitution ◽

Novel Mutation ◽

Factor Ix ◽

Causative Mutation ◽

Coding Region ◽

Body Of Knowledge ◽

Polymerase Chain ◽

Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

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