Advancement in Medicinal Plant Research

1-13

Authors: Hossein Tavallali, Atefeh Bahmanzadegan, Vahid Tavallali and Vahid Rowshan

Punica granatum L. is a native plant that is greatly dispensed in south of Iran. The massive pomace of Punica exhibits a challenging losses exposure difficulty for the processing industries. The resent study was aimed to investigate the composition and quality of P. granatum pomace and understand its bioactive compounds to introduce it to different industries such as pharmaceutical, food, medicinal, agricultural etcetera for optimum use. The SHS (static headspace) GC-MS on the Combi PAL system method was used to identify and analyze the volatile components (VCs). Four different extracts (60°C water, distilled water, methanol and methanol 80%) were analyzed for polyphenol compounds identified by HPLC-DAD, total flavonoid content (TFC), total phenolic content (TPC) and antioxidant capacity (DPPH(2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay). The relationship between TFC, TPC and antioxidant activity were statistically investigated. The dominant VCs of P. granatum were limonene (50.6%) followed by -pinene (25.1%), -terpinene (6.79%), -pinene (5.7%) and sabinene (2.59%), respectively. Gallic acid was major polyphenol compound in all studied extracts of marc. 60°C water extract from P. granatum pomace represented higher antioxidant capacity, TPC and TFC than other investigated extracts. This studied marc can be introduced to different industries as a source of bioactive and medicinal natural compounds because of its valuable phytochemical characterization.

85-90

Authors: Sleman Kadan, Yoel Sasson and Hilal Zaid

Portulaca oleracea L. is a grassy plant with a fleshy stem, succulent leaves, yellow or white small flowers and small black seeds. P. oleracea used as a medicinal plant in many countries, as a diuretic, febrifuge, vermifuge, antiseptic, anti-spasmodic, anti-bacterial, anti-inflammatory and anti-oxidant. It is in use also as anti-diabetic medicinal herb. The aim of this study was to evaluate the effect of the aerial parts of P. oleracea 50% ethanol/50% water and methanol based extracts on the glucose transporter-4 (GLUT4) translocation to the plasma membrane of muscle cells. Anti-diabetic activity of the extracts was examined by measuring the relative amount of GLUT4 translocation to the plasma membrane in the presence and absence of insulin. L6 muscle cells, stably expressing myc-tagged GLUT4, repeatedly shown to display insulin regulated GLUT4 traffic, were used as a model in this project. L6 myoblasts were pre-treated with increasing concentrations of P. oleracea extracts for 20 h, followed by 3 h serum-deprivation and insulin stimulation for 20 min and then GLUT4myc levels at the plasma membrane was determined. Toxicity of the extracts was determined by MTT assay and LDH leakage assay. Results obtained show no cytotoxic effects in cells treated for 24 h with 50% ethanol and methanol P. oleracea extract up to 1 mg/ml and 0.5 mg/ml extract respectively as measured with MTT and LDH-leakage assays. GLUT4 translocation to the plasma membrane was elevated by 2.6 and 4 folds (-/+ insulin) after treatment for 20 h with 1 mg/ml P. oleracea 50% ethanol extract and 1.7 and 3 folds (-/+ insulin) when treated with 0.5 mg/ml methanol extract. These results indicate that P. oleracea anti-diabetic proporties are mediated in part by increasing GLUT4 distribution in the muscle plasma membrane.

97-106

Authors: Mashael Al-Mutarrafi, Nagwa Thabet Elsharawy, Aisha Al-Ayafi, Ahmed Almatrafi and Hayam Abdelkader

The Kingdom of Saudi Arabia is considered one of the major dates producing zones in the world, contributing 13% of the total world production. Most of the dates are delivered to local markets without any treatment thus, may be leading to microbial contamination which may increase the incidence of microbial growth. Some pathogenic fungi associated with soft dates (Phoenix dactylifera, Linn.) collected from Almadinah Almunawarah, local markets, was isolated and molecularly identified during summer 2019. Contaminated date samples (Sukkary, Ajwa, Shalabi madinah, Barni and Sogea) were placed on PDA plates and incubated at 26°C for 5 days. Morphological and molecular identification of the resulting fungi were investigated. Molecular identification of fungal isolates was conducted using the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA). Fungal DNAs were amplified by PCR using ITS1-F (5’-TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’). The purified PCR products were sequenced and the obtained nucleotide sequences were submitted to GenBank on the NCBI. The results showed that the most dominant isolated fungi were Aspergillus niger (100%) associated with “Sogea”, followed by Aspergillus tubingensis (66%) isolated from “Shalabi Almadinah” (40%), Rhizopus stolonifer (36.3%) associated with “Ajwa”, Talaromyces minioluteus (14.2%) isolated from “Sukkary”, respectively. Soft dates are most likely contaminated due to the artificial rise in moisture content and the related inappropriate storage conditions. In conclusion, the study has declared that Aspergillus spp. considered the most fungal contamination of date fruits. Fungal contamination may have an effect on the public health by their mycotoxins which may lead to severe poisoning, diarrhea, emesis, and death. Most date fruits sold and marketed are neither covered nor protected in any way from environmental contamination. If these dates are consumed without thoroughly washing that may induce imminent mycotic infection. The study recommended that prevention of dates fruit mold spoilage should be avoided by application of hazard analysis, critical control point programs and adherence to hygienic good manufacturing practices.

68-78

Authors: Valentina Butleska Gjoroska, Marija Krstik, Lenka Cvetanovska and Liljana Koleva Gudeva

The role of alfalfa (Medicago sativa L.) in the development of agricultural production and intensification of forage production is due to the high potential of biomass on the one hand anti-oxidative and nutritional value on the other hand. Modern techniques have been used for analysing of organics acids. Organic acids are formed during cell respiration as products of total dissemination of carbohydrates and generally serve as starting substances for the biosynthesis of different compounds such as fatty acids and fats, amino acids, vitamins, coenzymes, pigments and other substances. Organic acids are key metabolites in plants, and their amounts depend on the intensity of cell respiration. The accumulation of some organic acids in plants is closely related to the intensity of enzymatic reactions as well as to various environmental factors. On the cellular level, the metabolism of organic acids is of high importance for several biochemical pathways, such as the cycle of tricarboxylic acids and the glyoxalate cycle. Organic acids that are synthesised in these cycles are used by plants as precursors in the biosynthesis of other organic compounds. Significant differences are found in the production of organics acids between the locations in Ovche Pole region on the one hand and Skopje and Tetovo regions on the other hand. We investigated various organic acids (citric, acetic, wine, malic and lactic acid) and the total acidity in alfalfa (Medicago sativa L.) from three different regions of the Republic of North Macedonia, in 19 different locations and three harvests.

38-43

Authors: Guilherme Perez Pinheiro, Maria Isabel Galbiatti, Mara Junqueira Carneiro and Alexandra Christine Helena Frankland Sawaya

Plectranthus amboinicus, also known as Indian borage or Mexican mint, is a species of Lamiaceae cultivated worldwide for ornamental, culinary and medicinal purposes, being used in folk medicine for the treatment of respiratory diseases and digestive problems. These properties are attributed to the abundance of phenolic monoterpenes present in its essential oil, which have been extensively described in the literature. The low yield of essential oil hinders further investigation, besides the extraction being a costly and time-consuming procedure. The application of headspace-solid-phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC-MS), an alternative technique in plant volatile analysis, is described in the present study for P. amboinicus growing in Brazil. Four SPME fibers with different coating composition were evaluated: polydimethylsiloxane (PDMS - 100 µm), carboxen/polydimethylsiloxane (CAR/PDMS - 75 µm), divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS - 50/30 µm) and polydimethylsiloxane/divinylbenzene (PDMS/DVB - 65 µm). PDMS fiber extracted a total of 19 identified compounds (52.46% of monoterpenes and 42.09% of sesquiterpenes), CAR/PDMS fiber 22 identified compounds (55.05% of monoterpenes and 25.51% of sesquiterpenes), DVB/CAR/PDMS fiber 24 identified compounds (67.22% of monoterpenes and 23.88% of sesquiterpenes) and PDMS/DVB fiber 26 identified compounds (64.3% of monoterpenes and 30.93% of sesquiterpenes). The extractions with PDMS/DVB and DVB/CAR/PDMS fibers resulted in similar chemical profiles when compared to essential oil composition, in particular the triple-coated DVB/CAR/PDMS fiber which extracted more compounds with high retention time. Both of these fibers are the most suitable in an untargeted analysis of P. amboinicus volatiles, while PDMS and CAR/PDMS may be suitable for target compounds or classes of compounds. The data reported in this study demonstrates that HS-SPME coupled to GC-MS is a rapid and simple method of analysis and compose a valuable tool for the study of P. amboinicus volatiles.

54-60

Authors: Jose Tapiero, Guillermo Salamanca and Carlos Marín

Aromatic plants make secondary metabolites that can be useful or toxic to an organism. In this work, we obtained Essential Oil (EO) by Microwave-assisted Hydrodistillation (MWHD). The process was carried out during the period from 15 min to 700 watts, with samples of 50, 100 and 200 g, collected in Vijes and Sevilla, Valle, Colombia. For each sample of essential oil the relative density at 20°C and the Refractive Index were evaluated (RI). The chemical composition was evaluated using GC-MS and its antiradical activity (AA) was carried out using the DPPH*, ABTS+ and ORAC methods. The process yield was between 0.087 ± 0.012 and 0.140 ± 0.005%. Significant differences were observed for the RI with values between 1.475 ± 0.001 and 1.484 ± 0.003 for a (Pv < 0.05). The highest contributions correspond to p-Cimeno (10.04), Terpinen-4-ol (9.48), Carvacrol (8.70%) and Thymol (1.95%) presented lowest values. The EO of oregano presents a good AA that ranges between 318.8 ± 0.009 to 320.6 ± 0.003 (Sevilla) and 310.8 ± 0.021 to 312.1 ± 0.014 (Vijes) mmol Trolox/100 g for DPPH*, and 23.852 ± 0.018 to 24.019 ± 0.011 (Sevilla) and 23.128 ± 0.012 to 23.412 ± 0.009 (Vijes) mmol Trolox/100g for ABTS+. Finally, oregano essential oils presented low molecular weight aliphatic compounds, being markers of these matrices.

1-7

Authors: Tsiba Gouollaly, Kevin Bikindou, Edmond Mabiangana, Aubin Nestor Loumouamou, Sarrah Boumba, Marie Claire Makambila, Gilles Figuérédo and Pierre Chalard

The chemical composition of the essential oil of Croton dibowskii Hutch from Congo-Brazzaville was analyzed by GC/FID and GC/SM. The chemical profile remains, for the most part, dominated by the cyperene (15.1%), borneol (12.5%), (6.9)-guaiadiene (8.2%), α-muurolene (7.9%) and 1.8-cineole (6.7%), representing in general 57.4% of sesquiterpene hydrocarbons and 23.8% of oxygenated monoterpne. We note in some respects, with a low content, the presence of monoterpenic hydrocarbons and sesquiterpenic oxygenated compounds of 5.0 and 3.0%, respectively. The antioxidant activity on the DPPH turns out to be very limited, it reaches inhibition percentage of 35% from the concentration of 2000 μg/ml with a CI50 estimated at 3000 μg/ml.

15-22

Authors: Kritiya Butthongkomvong, Araya Lukanawonakul, Sakunrat Tiewwanich, Pongsadhorn Pokpermdee, Isara Chiawiriyabunya and Tanawan Kummalue

The authors conducted a randomized, double-blind, placebo-controlled, pilot study to determine the efficacy and side effects of a well-known Thai herbal recipe in advanced hepatocellular carcinoma. Twenty advanced hepatocellular carcinoma patients were enrolled at Udon Thani Cancer Hospital, Thailand, between January 2015 and December 2017. They were divided into experimental and placebo groups, each with ten patients. The time to event was defined as the time to disease progression or death, and it was calculated using intention-to-treat analysis. Relative to the placebo group, the treatment group demonstrated a larger number of poor prognostic factors, such as larger tumor sizes, higher alpha-fetoprotein levels, worse Child–Pugh classifications, and much higher alkaline phosphatase levels. A Kaplan–Meier analysis revealed that the endpoint of the experimental group was not inferior to that of the control group, even though the experimental group had much worse prognostic factors. As to the safety of the herbal recipe, no adverse events were detected in the treatment group. The study demonstrated that the Thai herbal recipe might have some advantages for advanced hepatocellular carcinoma patients, especially in combination with supportive therapy.

31-37

Authors: Shah Zaman, Muhammad Zeeshan Bhatti, Du Hongmei and Shengquan Che

Climatic changes may affect soil quality of agricultural lands, especially by increasing salts deposition in the soil which results in soil salinity. Salinity stress affects several aspects of plant physiology, such as suppressing seed germination, reducing growth of roots and shoots, and altering growth regulators. Breeding salt tolerant plant cultivars has been proven inadequate to fully alleviate this problem. Salinity tolerance results from diverse mechanism, each controlled by a multi-gene system. Over the past decades, substantial progress has been made to elucidate the mechanisms underlying plant responses to mitigate adverse impacts of salinity. The overall problem is more tractable, if the plant response is dissected into diverse traits that are hypothesized to be involved in the tolerance to salinity. In this review, we comprehensively discuss plant adaption to salinity, the salt overly sensitive (SOS) pathway, ion homeostasis, determination of solutes and various molecular and proteomic approaches to understanding the salinity responses. This review provides response guidelines to enhance improvement of plant salt tolerance in combination with improved agricultural practices under saline conditions. Comprehensive understanding of all mechanisms for salinity stress response should be the ultimate of future studies.