Dinesh Chandra Agrawal

INFLUENCE OF VENTILATION CLOSURE, GELLING AGENT AND EXPLANT TYPE ON SHOOT BUD PROLIFERATION AND HYPERHYDRICITY IN SCROPHULARIA YOSHIMURAE - A MEDICINAL PLANT

SummaryWe report an improved procedure of in vitro propagation of Scrophularia yoshimurae—a medicinally important plant species indigenous to Taiwan. Induction of maximum shoot buds (22.75 per explant) was obtained with shoot tip explant cultured on Murashige and Skoog medium supplemented with 1.0mgl−1 benzyladenine (BA) and 0.2mgl−1 α-naphthaleneacetic acid and gelrite using dispense paper (DP) for ventilation closure of culture vessels. The type of gelling agents (agar and Gelrite) affected both quantity and quality of the shoots induced. Using aluminum foil for ventilation closure resulted in a higher number of hyperhydric shoots. Hyperhydricity was reduced by culturing shoots on a medium devoid of plant growth regulators in conjunction with the use of DP. Plantlet growth in vessels using DP was healthier and all plantlets survived after being transplanted to soil.

In vitro micropropagation and ex vitro acclimation of Bupleurum kaoi-an endangered medicinal plant native to Taiwan

SummaryThis study reports an improved protocol for in vitro-shoot multiplication and ex vitro acclimation of Bupleurum kaoi, an endangered medicinal herb. Nodal segments were cultured in half-strength Murashige and Skoog (MS) basal medium supplemented with different concentrations of benzyladenine (BA) and kinetin. The presence of 0.25 mg l−1 BA induced the highest number of shoots per explant after 8 wk of culture. Although BA was more effective than kinetin on shool multiplication, it induced hyperhydric shoots at all concentrations tested. The use of dispense paper (DP) instead of aluminum foil (AF) for container closure was found to reduce hyperhydricity and improve ex vitro acclimation. The best survival rate (61%) was obtained when plantlets were grown in MS basal medium containing 0.5 mg l−1 indole-3-butyric acid and 0.1–0.2 mg l−1 α-naphthaleneacetic acid using DP as container closure. Leaves of the plant treated with AF6 (two layers of AF as container closure and 6 wk of incubation) lacked epicuticular wax and possessed larger stomata, higher stomata density, and fewer functional stomata compared to those of plants treated with AF2+DP4 (two layers of AF for 2 wk, then replaced AF by three layers of DP for 4wk) and ex vitro-acclimated plantlets.

Development of an activation tagging system for the basidiomycetous medicinal fungus Antrodia cinnamomea.

This study describes the development of an efficient and reliable activation tagging system for the medicinal fungus Antrodia cinnamomea. For successful Agrobacterium tumefaciens-mediated transformation, different parameters were considered. The Agrobacterium concentration of 5 x 10(8) cfu ml(-1), 1mm acetosyringone, 25-d-old mycelia at 0.2 g ml(-1), and co-culture period of 6d were found to be the most optimal conditions for enhancing the transformation efficiency. The mitotic stability of transferred DNA (T-DNA) was demonstrated by growing eight randomly selected putative transformants in malt extract agar medium for five subcultures. Insertion of T-DNA into the genome of transformants was confirmed by PCR and Southern hybridization. Results showed that 88% of the mutants contained a single T-DNA insertion. Two of the mutants were observed with different triterpenoid profiles compared with the untransformed cultures. Our results suggest a new functional genomics approach to tag the triterpenoid biosynthesis genes in A. cinnamomea.

Molecular characterization of farnesyl pyrophosphate synthase from Bacopa monniera by comparative modeling and docking studies

Farnesyl pyrophosphate synthase (FPS; EC 2.5.1.10) is a key enzyme in isoprenoid biosynthetic pathway and provides precursors for the biosynthesis of various pharmaceutically important metabolites. It catalyzes head to tail condensation of two isopentenyl pyrophosphate molecules with dimethylallyl pyrophosphate to form C15 compound farnesyl pyrophosphate. Recent studies have confirmed FPS as a molecular target of bisphosphonates for drug development against bone diseases as well as pathogens. Although large numbers of FPSs from different sources are known, very few protein structures have been reported till date. In the present study, FPS gene from medicinal plant Bacopa monniera (BmFPS) was characterized by comparative modeling and docking. Multiple sequence alignment showed two highly conserved aspartate rich motifs FARM and SARM (DDXXD). The 3-D model of BmFPS was generated based on structurally resolved FPS crystal information of Gallus gallus. The generated models were validated by various bioinformatics tools and the final model contained only α-helices and coils. Further, docking studies of modeled BmFPS with substrates and inhibitors were performed to understand the protein ligand interactions. The two Asp residues from FARM (Asp100 and Asp104) as well as Asp171, Lys197 and Lys262 were found to be important for catalytic activity. Interaction of nitrogen containing bisphosphonates (risedronate, alendronate, zoledronate and pamidronate) with modeled BmFPS showed competitive inhibition; where, apart from Asp (100, 104 and 171), Thr175 played an important role. The results presented here could be useful for designing of mutants for isoprenoid biosynthetic pathway engineering well as more effective drugs against osteoporosis and human pathogens. Abbreviations IPP - Isopentenyl Pyrophosphate, DMAPP - Dimethylallyl Pyrophosphate, GPP - Geranyl Pyrophosphate, FPP - FPPFarnesyl Pyrophosphate, DOPE - Discrete Optimized Protein Energy, BmFPS - Bacopa monniera Farnesyl Pyrophosphate Synthase, RMSD - Root Mean square Deviation, OPLS-AA - Optimized Potentials for Liquid Simulations- All Atom, FARM - First Aspartate Rich Motif, SARM - Second Aspartate Rich Motif.

Andrographolide relieved pathological pain generated by spared nerve injury model in mice

Abstract Context: Andrographolide (Andro), found in large quantities in Andrographis paniculata Nees (Acanthaceae), is anti-inflammatory, especially in the central nervous system (CNS) glia. Objective: The objective of this study is to test Andro’s ability to reduce allodynia in a spared nerve injury model. Material and methods: Male 30 g BalbC mice were divided into four groups: (1) Sham-operated control (Sham-group); (2) nerve injured and treated with saline (Saline-group); (3) nerve injured and treated with Andro (Andro-group); (4) nerve injured and treated with non-steroidal anti-inflammatory drugs (NSAIDS) (NSAIDS-group). Andro or NSAIDS (diclofenac salt) were injected intraperitoneally at 5 mg/kg body weight daily. Mechanical allodynia was assessed by von Frey tests at 3, 7, and 14 d. For immunohistochemical analysis, samples were collected at 7 d. Results: The threshold for inducing allodynia increased and the response percentage reduced in the Andro-group when compared with the Saline-group, as well as when compared with NSAIDS groups throughout 3–14 d. The ratio of threshold for OP-Andro/OP-saline and for OP-Andro/OP-NSAIDS groups was 20.42 and 11.67 at 14 d, respectively. The ratio of response percentage for OP-Andro/OP-saline and for OP-Andro/OP-NSAIDS was 0.32 and 0.39 at 14 d, respectively. Interleukin-1 (IL-1) immunostaining in the spinal cord was reduced in the Andro-group. Astrocytic activities were not significantly reduced in the Andro-group compared with the Saline-group at 7 d post-operation (PO) Conclusions: Andro reduced mechanical allodynia more than NSAIDS at the same concentration, and the observed behaviour was associated with a reduction in inflammatory cytokine produced in the spinal cord.

Remedial education for high-risk university freshmen – the case of a university of technology in Taiwan

Abstract Potential high-risk freshmen for three core courses (BasicMath, Calculus, and Computing) in the university were identified based on the “College Students’ Adjustment Check List (CSACL)” data available with the Student Development Centre in the Office of Students’ Affairs of the university. The study demonstrates that to ameliorate the problem of unpreparedness of freshmen, to check failure rates, and efficient use of limited resources, an effective remedial system could be developed by the combined inputs from the Office of Students’ Affairs, Computer Centre, Academic Affairs, and the Institutional Research. Our study corroborates the findings of other researchers that gender, teacher, department, high school performance, enrolment channel, and loads during remedial course influence the learning outcome. Also, it was found that the use of multi-evaluation approaches in different enrolment channels can attain higher retention rates.

Biotechnology of Medicinal Plants and Fungi in Taiwan: Production of Bioactive Secondary Metabolites in In Vitro Culture Systems

Plants and fungi provide a vast range of natural products including pharmaceuticals with diverse chemical structures and a broad array of biological activities. In the past few decades, there has been a worldwide resurgence of interest in the study and use of medicinal plants and fungi in the health-care system. Several medicinal plants and fungi are involved in the industrial processing of profitable products used in human medicines. However, due to severe constraints on the availability of desired plant and fungus materials, different strategies, including tissue culture, have been extensively studied. The present chapter focuses on the application of various in vitro culture systems in the production of bioactive secondary metabolites in medicinal herbs and fungi in Taiwan. All the species described in this article are sources of traditional Chinese medicines. The review article includes the research work carried out in our laboratory on the production of plant secondary metabolites in callus cultures of Saussurea involucrata, Solanum melongena, and Salvia miltiorrhiza and in cell suspension cultures of Taxus mairei, Gnetiana davidii var. formosana, and Angelica dahurica var. formosana. Also, plant metabolites were obtained from in vitro shoots of Glossogyne tenuifolia, Saussurea involucrata, and Polygonum multiflorum, aerial parts of Scrophularia yoshimurae, tubers of Corydalis yanhusuo, tissue culture plants, and hairy roots of Gentiana scabra. Also, we could achieve the production of terpenoids in Antrodia cinnamomea, a medicinal fungus, and tanshinones in Salvia miltiorrhiza using T-DNA activation-tagging technique (Agrobacterium tumefaciens-mediated transformation).

Mushrooms: A Pandora Box of Cardioprotective Phytochemicals

Overwhelming consensus among medical authorities and scholarly bodies on the high susceptibility to chronic ailments such as coronary diseases, cancers, and diabetes and the failure to make any leap forward progress in controlling casualties or even to completely understand their pathology is a frightening reality. To comprehend alterations, additions, and management of diet is a preferable approach not only to prevent the occurrence of cardiovascular diseases but also to precise and enhance treatment measures. Proper application of potential drugs is possible only by establishing a systemic correlation and compilation of the knowledge obtained on the possible bioactive drugs. In this perspective gathering knowledge on the health-promoting potential of mushrooms which are considered as one of the promising sources of potential products that provide cardioprotection is indispensable. While there are several mushrooms traditionally utilized around the world for the treatment of cardiovascular diseases (CVD), they are also being cautiously evaluated experimentally for the available evidences of ethnopharmacology. Some therapeutic mushrooms have preclinical studies to demonstrate that uptake of these organic dietary supplements and their constituents as a therapeutic alternative or supplement is conceivable, and further evaluations are carried out to help in lessening the prevalence and mortality of CVD by incorporating them either as a population medicine or as a clinical medicine. A few examinations have demonstrated the effect of mushrooms and their bioactive compounds on metabolic markers such as low-density lipoprotein, high-density lipoprotein, total cholesterol, fasting triacylglycerol, and homocysteine levels and on conditions such as hypertension, body hemostasis, oxidative stress, and inflammation which are associated with cardiovascular ailments. The focus of this chapter will primarily be on mushrooms used traditionally for the treatment of CVD.

Medicinal Plants and Fungi: Recent Advances in Research and Development

Ayurveda is one of the ancient traditional healthcare systems that originated in India. A number of herbal-based medicinal preparations have been used for the treatment of health disorders associated with the nervous system. According to Alzheimer’s disease Facts and Figures, millions of people around the world are suffering with cognitive impairment. Cognitive ailments and diseases are a group of disorders associated with mental health. The cognitive disorders mainly comprise of acute and chronic or reversible or irreversible conditions such as amnesia, delirium, and various types of dementia. These disorders primarily cause deficits in cognitive tasks associated with awareness, insight, knowledge, memory, and problem-solving skills. Alzheimer’s disease is the most common type of dementia. It is a chronic neurodegenerative disorder that occurs due to excessive protein deposition inside and outside the neuron, oxidative stress, apoptosis, mitochondrial dysfunction, inflammation, and excitotoxicity. These neurotoxic mechanisms cause synaptic disturbance, alteration of neurotransmission leading

Salvia miltiorrhiza: A Medicinal Herb from Metabolites to Pathway Engineering

The dried, red-colored roots of Salvia miltiorrhiza (Lamiaceae) also called “danshen” in colloquial language in China have been used in traditional Chinese medicine for thousands of years to treat hypertension and other cardiovascular ailments. The main constituents of danshen are hydrophilic phenolic acids and lipophilic tanshinones. The various uses of danshen in traditional as well as modern medicines have motivated an intensive research on compounds in S. miltiorrhiza. In recent years, more than 110 compounds have been isolated from S. miltiorrhiza and their structure was identified. Tanshinones and their derivatives have been demonstrated to possess properties of slowing down or curing various ailments related to cardiovascular, cerebrovascular, respiratory, liver, nervous system, cancer, Alzheimer’s, and Parkinson’s diseases. With the increasing demand of this herb, an unrestricted collection to supply raw materials and the extraction of its constituents have severely threatened the natural habitats of S miltiorrhiza. This has prompted the researchers to develop alternative strategies for metabolite production. Several in vitro methodologies have been established to generate callus, cell suspension culture, hairy roots, and plant regeneration. Different regulators and elicitors for plant growth have been employed to enhance levels of different constituents. The advent of sequencing technologies, whole genome, and expression data has helped to provide insights and identification of pathway genes involved in the biosynthesis. This book chapter gives a brief description of in vitro methodologies, use of different elicitors, gene functions, genetic modifications, expression profiling for a better understanding, and enhancement of the constituents in S. miltiorrhiza.