Effect of salinity stress and surfactant treatment with zinc and boron on morpho-physiological and biochemical indices of fenugreek (Trigonella foenum-graecum).

Micronutrient application has a crucial role in mitigating salinity stress in crop plants. This study was carried out to investigate the effect of zinc (Zn) and boron (B) as foliar applications on fenugreek growth and physiology under salt stress (0 and 120 mM). After 35 days of salt treatments, three levels of zinc (0, 50, and 100 ppm) and two levels of boron (0 and 2 ppm) were applied as a foliar application. Salinity significantly reduced root length (72.7%) and shoot length (33.9%), plant height (36%), leaf area (37%), root fresh weight (48%) and shoot fresh weight (75%), root dry weight (80%) and shoot dry weight (67%), photosynthetic pigments (78%), number of branches (50%), and seeds per pod (56%). Fenugreek's growth and physiology were improved by foliar spray of zinc and boron, which increased the length of the shoot (6%) and root length (2%), fresh root weight (18%), and dry root weight (8%), and chlorophyll a (1%), chlorophyll b (25%), total soluble protein content (3%), shoot calcium (9%) and potassium (5%) contents by significantly decreasing sodium ion (11%) content. Moreover, 100 ppm of Zn and 2 ppm of B enhanced the growth and physiology of fenugreek by reducing the effect of salt stress. Overall, boron and zinc foliar spray is suggested for improvement in fenugreek growth under salinity stress.

Tissue-specific variations of piperine in ten populations of Piper longum L.: bioactivities and toxicological profile.

P. longum L., one of the most significant species of the genus Piperaceae, is most frequently employed in Indian-Ayurvedic and other traditional medicinal-systems for treating a variety of illnesses. The alkaloid piperine, is the key phytoconstituent of the plant, primarily responsible for its' pharmacological-impacts. The aim of the study is to analyse the intra-specific variation in piperine content among different chemotypes (PL1 to PL 30) and identify high piperine yielding chemotype (elite-chemotype) collected from 10 different geographical regions of West Bengal by validated HPTLC chromatography method. The study also focused on the pharmacological-screening to better understand the antioxidant activity of the methanol extracts of P. longum by DPPH and ABTS radical-scavenging activity and genotoxic activity by Allium cepa root tip assay. It was found that the P. longum fruit chemotypes contain high amount piperine (highest 16.362 mg/g in chemotype PL9) than the stem and leaf chemotypes. Both DPPH and ABTS antioxidant assays revealed that P. longum showed moderate radical-scavenging activity and the highest activity was found in PL9 (fruit) chemotype with IC50 values of 124.2 ± 0.97 and 104 ± 0.78 µg/ml respectively. The A. cepa root tip assay showed no such significant genotoxic-effect and change in mitotic-index. The quick, reproducible, and validated HPTLC approach offers a useful tool for determining quantitative variations of piperine among P. longum chemotypes from different geographical-regions and also according to the different tissues and choose elite genotypes with high piperine production for continued propagation and commercialization for the pharmaceutical sector. Additionally, the plant's in-vitro antioxidant property and lack of genotoxicity directly supports its' widespread and long history of use as a medicinal and culinary plant.

Quantification of Tissue-Specific Paclitaxel in Himalayan Yew Using HPTLC-Densitometric Analysis, Assessment of Toxicological Activity, and Tissue-Specific Evaluation of Antioxidant Activity.

Taxus wallichiana Zucc., commonly known as the Himalayan Yew, is currently experiencing endangerment due to excessive harvesting and sluggish growth resulting from the extraction of paclitaxel, a crucial plant-derived medication employed in the treatment of cancer. T. wallichiana contains various phytochemicals, including paclitaxel, a diterpenoid that has been utilized as an anticancer medication. In order to extract paclitaxel while maintaining the species' survival, it is difficult to determine the most effective plant parts. We determined the diterpenoid paclitaxel content using modern analytical methods such as high-performance thin-layer chromatography-densitometric analysis. Furthermore, toxicological evaluations were carried out and tissue-specific antioxidant activity was statistically analyzed using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric reducing antioxidant power (FRAP), Folin-Ciocâlteu (FC), and 2,2-diphenyl-ß-picrylhydrazyl (DPPH) assays. The results of our study offer significant contributions to the identification of optimal plant components for the extraction of paclitaxel. This information is crucial in the conservation of T. wallichiana and in mitigating the difficulties associated with its threatened classification. The present investigation makes a valuable contribution toward the advancement of sustainable methodologies in the manufacturing of paclitaxel, as well as the preservation of T. wallichiana for posterity. Bark exhibited the maximum paclitaxel yield at a content of 29162.3 µg/g dry weight. The accuracy of the method has been validated in accordance with the guidelines outlined by the International Council for Harmonisation (ICH). The current investigation evaluated the potential cytotoxic and genotoxic effects of the aqueous extracts on meristematic cells from the roots ofAllium cepa. The extracts obtained from the bark exhibited noteworthy cytotoxic and mitotic characteristics. The current investigation holds potential significance for the pharmaceutical sector in terms of identifying superior chemotypes of T. wallichiana that produce high levels of paclitaxel. Conducting a toxicological assessment on various tissues of T. wallichiana chemotypes through employment of the Allium cepa test would facilitate the identification of any potential genotoxic characteristics. The present study aimed to investigate four distinct assays, namely, DPPH, ABTS, FRAP, and FC, for the evaluation of the antioxidant potential of diverse T. wallichiana plant extracts and standard substances. The findings suggest that FRAP and ABTS exhibit a strong correlation. In general, the entirety of the tissue extract exhibited commendable antioxidant capacity, thereby rendering it a promising contender for diverse applications.

Opium alkaloids, biosynthesis, pharmacology and association with cancer occurrence.

Papaver somniferum L. (Family: Papaveraceae) is a species well known for its diverse alkaloids (100 different benzylisoquinoline alkaloids (BIAs)). L-tyrosine serves as a precursor of several specific metabolites like BIAs. It has been used as an antitussive and potent analgesic to alleviate mild to extreme pain since ancient times. The extraction of pharmaceutically important alkaloids like morphine and codeine from poppy plant reflects the need for the most suitable and standard methods. Several analytical and extraction techniques have been reported in open literature for morphine, codeine and other important alkaloids which play a vital function in drug development and drug discovery. Many studies suggest that opioids are also responsible for adverse effects or secondary complications like dependence and withdrawal. In recent years, opium consumption and addiction are the most important risk factors. Many evidence-based reviews suggest that opium consumption is directly linked or acts as a risk factor for different cancers. In this review, we highlight significant efforts related to research which have been done over the past 5 decades and the complete information on Papaver somniferum including its phytochemistry, pharmacological actions, biosynthetic pathways and analytical techniques of opium alkaloid extraction and the link between opium consumption and cancer-related updates.

Assessment of knowledge, attitude, and practices of tuberculosis patients towards DOTs regimen in Jimma health center, Jimma zone, southwest Ethiopia.

Objectives: The purpose of conducting this study was to assess the knowledge, attitude, and practices of tuberculosis patients towards dots regimen in Jimma health Center. In addition, assessing the attitude of tuberculosis towards factors, modern, and traditional medicine was another aim of this study. Methods: A cross-sectional study was conducted based on patient card and registration book at TB follow-up clinic at Jimma health center, southwest Ethiopia. The study population comprises 150 participants, who were selected from the patient registration book by interviewing and questionnaires. Additionally, cross-tabulation and OR with 95% confidence interval were computed. Results: In this study, only 103 (68.7%) and 47 (31.3%) patients scored higher and lower knowledge status respectively. Of the total enrolled governmental employee, 25 (16.7 %) patients had an overall high knowledge status. On the other hand, 101 (67.3%) and 49 (32.7 %) patients scored high and low overall attitude respectively. Conclusions: In this study, the knowledge and attitude of patients about TB and its treatment were found within a low level of perception. Therefore, implementation of health education and awareness creation through different mobilization mechanisms are required to avoid the problem for patients.

Optimization of harvest and extraction factors by full factorial design for the improved yield of C-glucosyl xanthone mangiferin from Swertia chirata.

Swertia chirata Buch.-Ham. ex C.B. Clarke is an important medicinal plant used in various herbal formulations as it shows significant biological activities such as hepatoprotective, hypoglycemic, anti-inflammatory, antimalarial, antioxidant and anti-parkinson. C-glucosyl xanthone glycoside (mangiferin) is known as bio-marker compound of genus Swertia L. Development of efficient extraction methods of C-glucosyl xanthone mangiferin from Swertia chirata was attempted by optimizing the pre-harvest, post-harvest and extraction techniques by full factorial design. Firstly, a full factorial design was implemented to evaluate the single and interactive effects of pre-harvest (growth stage and plant part), post-harvest (drying condition and storage periods) followed by selection of best extraction technique such as heat reflux extraction (HRE), microwave assisted extraction (MAE) and ultrasound assistant extraction (UAE) at different solvent types on mangiferin yield. HPTLC and HPLC techniques were used for the determination of mangiferin content in extracts generated from different plant samples. In addition, anti-oxidant and anti-diabetic properties were determined by using DPPH assay and percentage inhibition of α­amylase enzyme. Substantial variation of mangiferin yield, ranged from 1.46 to 4.86% was observed, depending on the growth stage, plant part, drying condition, storage periods and extraction method. Results showed that drying of the leaves of Swertia chirata in the shade harvested at budding stage and stored for not more than 1 month was recommended for obtaining a higher mangiferin yield. Among different extraction techniques, MAE and UAE in 50% aqueous ethanol solvent were found to be efficient and cost-effective with better yield of mangiferin (4.82% and 4.86%, respectively) as compared to HRE (4.14%). Highest DPPH activity and percentage inhibition of α­amylase was observed in the aqueous ethanol extract of S. chirata leaves harvested at bud-stage of plant followed by flowering stage. The study shows that optimization of various factors by full factorial design was found to be an effective procedure to improve mangiferin yield from Swertia chirata and can be used for extraction of mangiferin.

Optimization of diosgenin extraction from Dioscorea deltoidea tubers using response surface methodology and artificial neural network modelling.

INTRODUCTION: Dioscorea deltoidea var. deltoidea (Dioscoreaceae) is a valuable endangered plant of great medicinal and economic importance due to the presence of the bioactive compound diosgenin. In the present study, response surface methodology (RSM) and artificial neural network (ANN) modelling have been implemented to evaluate the diosgenin content from D. deltoidea. In addition, different extraction parameters have been also optimized and developed. MATERIALS AND METHODS: Firstly, Plackett-Burman design (PBD) was applied for screening the significant variables among the selected extraction parameters i.e. solvent composition, solid: solvent ratio, particle size, time, temperature, pH and extraction cycles on diosgenin yield. Among seven tested parameters only four parameters (particle size, solid: solvent ratio, time and temperature) were found to exert significant effect on the diosgenin extraction. Moreover, Box-Behnken design (BBD) was employed to optimize the significant extraction parameters for maximum diosgenin yield. RESULTS: The most suitable condition for diosgenin extraction was found to be solid: solvent ratio (1:45), particle size (1.25 mm), time (45 min) and temperature (45°C). The maximum experimental yield of diosgenin (1.204% dry weight) was observed close to the predicted value (1.202% dry weight) on the basis of the chosen optimal extraction factors. The developed mathematical model fitted well with experimental data for diosgenin extraction. CONCLUSIONS: Experimental validation revealed that a well trained ANN model has superior performance compared to a RSM model.

Screening the elite chemotypes of Gloriosa superba L. in India for the production of anticancer colchicine: simultaneous microwave-assisted extraction and HPTLC studies.

BACKGROUND: Gloriosa superba L. (Colchicaceae) is a high-value medicinal plant indigenous to Africa and Southeast Asia. Its therapeutic benefits are well-established in traditional medicines including Ayurveda. It is well known for its natural bioactive compound colchicine which exhibits a wide range of pharmacological activities i.e. rheumatism, gout and was also introduced into clinical practices. The increasing demand as well as its illegal harvesting has brought this valuable plant under threatened category. METHODS: The present investigation describes a microwave assisted extraction (MAE) strategy coupled with a densitometric-high performance thin layer chromatographic (HPTLC) methodology for the analysis of colchicine from 32 different populations of G. superba. A Box-Behnken statistical design (3 level factor) has been employed to optimize MAE, in which power of microwave, time of irradiation, aqueous ethanol and pH were used as independent variables whereas colchicine was used as the dependent variables. Chromatography was carried out on Silica gel 60 F254 TLC plates with toluene: methanol, 85:15 (v/v) being used as solvent system. Densitometric measurement was performed at λ=254 nm following post-derivatization (10% methanolic sulphuric acid). RESULTS: Optimal conditions for extraction to obtain the maximum colchicine yield was found to be 7.51 mg g- 1 which was very close to be predicted response 7.48 mg g- 1 by maintaining microwave power (460 W), irradiation time (6.4 min), aqueous ethanol-30, pH -3. Colchicine content ranged between 2.12-7.58 mg g- 1 among 32 G. superba populations in which only three chemotypes viz. GS- 1, GS- 3, and GS- 2 collected from West Bengal and Sikkim, respectively exhibited maximum yield of colchicine. CONCLUSION: Therefore, this newly developed optimized MAE coupled with HPTLC densitometry methodology not only quantifies colchicine in order to identify elite chemotypes of G. superba, but it also encourages in selecting high yielding populations of the plants for industrial use and economic boost for the farmers. This validated, simple and reproducible HPTLC protocol is being used for the first time to estimate colchicine from natural populations of G. superba obtained from 32 different geographical regions of India.

An evidence based efficacy and safety assessment of the ethnobiologicals against poisonous and non-poisonous bites used by the tribals of three westernmost districts of West Bengal, India: Anti-phospholipase A2 and genotoxic effects.

INTRODUCTION: To explore the ethnobiological wisdom of the tribals of three western districts of West Bengal, India against poisonous and non-poisonous bites and stings, a quantitative approach was adopted. These age-old yet unexplored knowledge can be utilized in finding lead-molecules against poisonous and non-poisonous animal-bites. Further, an evidence-based approach is needed to assess the venom-neutralization ability of plants by experimental studies. MATERIALS AND METHODS: During 2008-2009 and 2012-2017, 11 ethnomedicinal surveys were carried out to explore the use of medicinal flora and fauna via conducting open semi-structured interviews with 47 traditional healers (THs) or informants. The retrieved dataset was statistically evaluated using seven quantitative-indexes: use-value (UV), informants'-consensus-factor (ICF), fidelity-level (FL), relative-importance (RI), cultural importance-index (CI), index of agreement on remedies (IAR) and cultural agreement-index (CAI). Anti-phospholipaseA2 (PLA2) properties of selected plant extracts were also examined. In addition, the cytotoxicity and genotoxicity of the water extract of the plants showing high FL as well as significant PLA2 inhibitory potential were investigated using Allium cepa root tip assay. RESULTS: A total of 41 traditional-formulations (TFs) containing 40 plant species (of 39 genera from 28 families) and 3 animal species were prescribed by the THs. Fabaceae exhibited most number of medicinal plants. Piper nigrum (1.78) and Apis cerana indica and Crossopriza lyoni (both 0.21) exhibited the highest UV among the plants and the animals respectively. Stinging of centipede and dog/cat/hyena bite displayed highest ICF (1.00 each). Among the plants, the maximum RI (0.91) and CI (4.98) values were observed for Aristolochia indica. IAR (1.00) was recorded maximum for Achyranthes aspera, Gloriosa superba, Lycopodium cernuum, Smilax zeylanica and Streblus asper. Maximum CAI value was noted for Piper nigrum (5.5096). Among the animals, Apis cerana indica (0.31) and Crossopriza lyoni (1.52) displayed the highest RI and CI values respectively. Crossopriza lyoni (0.99) and Apis cerana indica (1.3871) exhibited maximum IAR and CAI values respectively. Plants showing higher FL exhibited higher anti-PLA2 activity via selective inhibition of human-group PLA2. In addition, Allium cepa root tip assay has indicated the safety and/or toxicity of the plant parts prescribed by the THs. Root water extracts of Aristolochia indica and Gloriosa superba exhibited significant genotoxicity and cytotoxicity. CONCLUSIONS: Three western districts of West Bengal is the natural abode for many tribal and non-tribal communities. A noteworthy correlation was established between the plants used against poisonous-bites and their anti-PLA2 activity. A few plant parts used by the THs also exhibited high toxicity. Such alternative medical practices serve as the only option in these underprivileged and backward areas during medical-exigencies.

Validation and quantification of major biomarkers in 'Mahasudarshan Churna'- an ayurvedic polyherbal formulation through high-performance thin-layer chromatography.

BACKGROUND: Mahasudarshan Churna (MC) is a polyherbal Ayurvedic medicine that is employed in fever (especially chronic type), cold and malaria, improvement of digestion and appetite, removes toxins from the blood, boosts immunity and protects against common bacterial infections. METHODS: Validation and quantification of oleanolic acid (OA), ursolic acid (UA), mangiferin (M), gallic acid (GA), quercetin (Q) and curcumin (C) in commercial MC formulations by HPTLC method. Mobile phase, hexane: ethyl acetate: acetone (16.4: 3.6: 0.2, v/v) was used for the separation of OA and UA; ethyl acetate: glacial acetic acid: formic acid: water (20: 2.2: 2.2: 5.2 v/v) for the development of M; and toluene: ethyl acetate: formic acid (13.5: 9: 0.6 v/v) for the separation of GA, Q and C in crude sample extracts. Visualization and scanning were performed at λ = 530 nm for OA and UA, at λ = 254 nm for M and at λ = 366 nm for GA, Q and C. In addition, HPLC-PDA analysis was used to confirm the HPTLC results. RESULTS: Major bio-active compounds in MC formulations were oleanolic acid (1.54-1.78%), mangiferin (1.38-1.52%) and gallic acid (1.01-1.15%); followed by ursolic acid (0.79-0.98%), curcumin (0.45-0.67%) and quercetin (0.22-0.34%). CONCLUSION: Analysis of bio-active compounds in the present study was performed using HPTLC methods and later HPTLC results were compared with HPLC. These two methods give comparable results and there was no statistically significant difference between the mean values for all extracts. Present study concluded that this HPTLC technique is low cost, fast, precise, and accurate which can be employed for the quantification of xanthonoid (M), triterpenoids (OA, UA) and phenolics (GA, Q and C) in samples/formulations. Furthermore, present HPTLC method can be conveniently employed for routine quality control analysis of all the six marker compounds in marketed Ayurvedic/herbal formulations.

Evaluation of antioxidant, antibacterial, and antidiabetic potential of two traditional medicinal plants of India: Swertia cordata and Swertia chirayita.

BACKGROUND: Swertia cordata and Swertia chirayita are temperate Himalayan medicinal plants used as potent herbal drugs in Indian traditional systems of medicine (Ayurvedic, Unani and Siddha). OBJECTIVE: Assessment of Antioxidant, antibacterial, and antidiabetic potential of Swertia cordata and Swertia chirayita. MATERIALS AND METHODS: Phytochemicals of methanolic and aqueous extracts of the two Swertia species were analyzed. The antioxidant potential of all the extracts was assessed by measuring total phenolic content, total flavonoid content and free radical scavenging potential was assessed by 1,1-diphenyl-2-picrilhydrazyl (DPPH) assay, antibacterial activity was assessed against various pathogenic and nonpathogenic bacteria in vitro by Kirby-Bauer agar well diffusion method and antidiabetic activity was assessed by α-amylase inhibition. RESULTS: Methanolic leaf extracts of both the species of Swertia contain significant antibacterial as well as anti-diabetic potential, whereas methanolic root extracts of both species were found to have potential antioxidant activity. However, Swertia chirayita showed better activities than Swertia cordata although both species have good reputation in traditional Indian medicine. CONCLUSION: Both the species are having high medicinal potential in terms of their antioxidant, antibacterial and antidiabetic activities. Studies are required to further elucidate antioxidant, anti-diabetic and antibacterial potentials using various in-vitro, in-vivo biochemical and molecular biology techniques.

Improved growth and colchicine concentration in Gloriosa superba on mycorrhizal inoculation supplemented with phosphorus-fertilizer.

BACKGROUND: Gloriosa superba produces an array of alkaloids including colchicine, a compound of interest in the treatment of various diseases. The tuber of Gloriosa superba is a rich source of colchicine which has shown anti-gout, anti-inflammatory, and anti-tumor activity. However, this promising compound remains expensive and Gloriosa superba is such a good source in global scale. Increase in yield of naturally occurring colchicine is an important area of investigation. MATERIALS AND METHODS: The effects of inoculation by four arbuscular mycorrhizal (AM), fungi, Glomus mossae, Glomus fasciculatum, Gigaspora margarita and Gigaspora gilmorei either alone or supplemented with P-fertilizer, on colchicine concentration in Gloriosa superba were studied. The concentration of colchicine was determined by high-performance thin layer chromatography. RESULTS: The four fungi significantly increased concentration of colchicine in the herb. Although there was significant increase in concentration of colchicine in non-mycorrhizal P-fertilized plants as compared to control, the extent of the increase was less compared to mycorrhizal plants grown with or without P-fertilization. This suggests that the increase in colchicine concentration may not be entirely attributed to enhanced P-nutrition and improved growth. Among the four AM fungi Glomus mossae was found to be best. The total colchicine content of plant (mg / plant) was significantly high in plants inoculated with Glomus mossae and 25 mg kg(-1)phosphorus fertilizer (348.9 mg /plant) while the control contain least colchicine (177.87 mg / plant). CONCLUSION: The study suggests a potential role of AM fungi in improving the concentration of colchicine in Gloriosa superba tuber.