Jones O. Moody

Cyclotide Discovery in Gentianales Revisited—Identification and Characterization of Cyclic Cystine-Knot Peptides and Their Phylogenetic Distribution in Rubiaceae Plants

Cyclotides are a unique class of ribosomally synthesized cysteine-rich miniproteins characterized by a head-to-tail cyclized backbone and three conserved disulfide-bonds in a knotted arrangement. Originally they were discovered in the coffee-family plant Oldenlandia affinis (Rubiaceae) and have since been identified in several species of the violet, cucurbit, pea, potato, and grass families. However, the identification of novel cyclotide-containing plant species still is a major challenge due to the lack of a rapid and accurate analytical workflow in particular for large sampling numbers. As a consequence, their phylogeny in the plant kingdom remains unclear. To gain further insight into the distribution and evolution of plant cyclotides, we analyzed ∼300 species of >40 different families, with special emphasis on plants from the order Gentianales. For this purpose, we have developed a refined screening methodology combining chemical analysis of plant extracts and bioinformatic analysis of transcript databases. Using mass spectrometry and transcriptome-mining, we identified nine novel cyclotide-containing species and their related cyclotide precursor genes in the tribe Palicoureeae. The characterization of novel peptide sequences underlines the high variability and plasticity of the cyclotide framework, and a comparison of novel precursor proteins from Carapichea ipecacuanha illustrated their typical cyclotide gene architectures. Phylogenetic analysis of their distribution within the Psychotria alliance revealed cyclotides to be restricted to Palicourea, Margaritopsis, Notopleura, Carapichea, Chassalia, and Geophila. In line with previous reports, our findings confirm cyclotides to be one of the largest peptide families within the plant kingdom and suggest that their total number may exceed tens of thousands.

Uterine contractility of plants used to facilitate childbirth in Nigerian ethnomedicine

Ethnopharmacological relevance Pregnant women in Nigeria use plant preparations to facilitate childbirth and to reduce associated pain. The rationale for this is not known and requires pharmacological validation. Aim of study Obtain primary information regarding the traditional use of plants and analyze their uterine contractility at cellular level. Materials and methods Semi-structured, open interviews using questionnaires of traditional healthcare professionals and other informants triggered the collection and identification of medicinal plant species. The relative traditional importance of each medicinal plant was determined by its use-mention index. Extracts of these plants were analyzed for their uterotonic properties on an in vitro human uterine cell collagen model. Result The plants Calotropis procera, Commelina africana, Duranta repens, Hyptis suaveolens, Ocimum gratissimum, Saba comorensis, Sclerocarya birrea, Sida corymbosa and Vernonia amygdalina were documented and characterized. Aqueous extracts from these nine plants induced significant sustained increases in human myometrial smooth muscle cell contractility, with varying efficiencies, depending upon time and dose of exposure. Conclusion The folkloric use of several plant species during childbirth in Nigeria has been validated. Seven plants were for the first time characterized to have contractile properties on uterine myometrial cells. The results serve as ideal starting points in the search for safe, longer lasting, effective and tolerable uterotonic drug leads.

Use of medicinal plants for the treatment of measles in Nigeria.

AIM OF THE STUDY The present study was an ethnobotanical survey of three Local Government areas of the Ijebu area of Ogun State in southwest Nigeria for plants used in the treatment of measles. MATERIALS AND METHODS Unstructured interviews were conducted among both urban and rural dwellers of three major groups of Ijebu people inhabiting the area (Ijebu North, Ijebu northeast and Ijebu Ode Local Governments). RESULTS A total of 20 respondents constituted by herbalists, herbsellers and old people that have privileged information on the plants used in the treatment of measles among children were encountered during the survey. Twenty-three plant species belonging to 18 Angiosperm families were said to possess curative properties for the cure of measles among the local populace. Amongst the most frequently used plants are Elytraria marginata Vahl, Peperomia pellucida (L.) Humb., Bonpl. & Kunth, Vernonia amygdalina Del., Momordica charantia L., Newbouldia laevis (P. Beauv.) Seem. ex Bureau, and Ocimum gratissimum L. CONCLUSION The most frequently mentioned family is Cucurbitaceae. The mode of preparation and recommended dosages are enumerated in this paper. The results of the study call for an urgent need of the introduction of a strategy for the conservation of indigenous medicinal plants in the area.

Combination Studies of Eucalyptus torelliana F. Muell. Leaf Extracts and Clarithromycin on Helicobacter pylori

Helicobacter pylori is a Gram‐negative bacillus that is associated with the development of gastritis and peptic ulcer disease (PUD). In Nigeria, leaf extracts of Eucalyptus torelliana F. Muell. are used in traditional medicine to treat PUD and other gastrointestinal ailments. The additive and synergistic effects of E. torelliana leaf extracts, in combination with clarithromycin, were investigated using two types of H. pylori strains (ATCC 43629, ATCC 43579) and four clinical isolates of H. pylori (Ed, A2, G1‐1, 5514) in the checkerboard assay and the fractional inhibitory concentration (FIC) index. A time–kill study was also performed on the strain ATCC 43579. The results showed that the E. torelliana extract inhibited the growth of all H. pylori strains, and the addition of one of the isolated active compounds, namely compound 2 (a substituted pyrenyl ester) enhanced the activity of clarithromycin. The minimum inhibitory concentration values of clarithromycin and the botanical compound were reduced twofold (from 0.125 to 0.0625 µg/mL and > 100 to 50 µg/mL respectively). A 100% reduction in CFU/mL of H. pylori ATCC 43579 was observed with the combination of 0.25 µg/mL clarithromycin and 100 µg/mL and 200 µg/mL compound 2 after 3 h of exposure. The results of the investigation showed that the combination of botanical compounds and antibiotics may be beneficial in the treatment of H. pylori infections. Copyright

Traditional anti-fever phytotherapies in Sagamu and Remo North Districts in Ogun State, Nigeria.

An ethnobotanical survey was conducted to identify the medicinal plants in Sagamu Local Government Area (LGA), and Remo North LGA of Ogun State, Nigeria, implicated in the treatment of various types of fever. The survey tools consisted of a semi-structured questionnaire and oral interviews. Seventy respondents mostly ages 31 to 50 covered in this survey were drawn from among the villagers, herbalists, herb sellers, traditional medical practitioners, students, community leaders, and elders who are mostly educated. Four types of fever including malaria, yellow fever, typhoid, and cold were identified with malaria and yellow fever being very common. The majority of the respondents were quite knowledgeable in the etiology, symptoms, and seasonality of all fevers except cold. Survey results document 116 anti-fever herbal recipes being administered as oral decoction or infusion for both curative and preventive purposes. Treatment is usually devoid of known side effects. Cymbopogon citratus, Citrus aurantifolia, Enantia chlorantha, Carica papaya, Morinda lucida, and Lawsonia inermis were frequently included in anti-fever herbal recipes.

Ethnobotanical survey of Rinorea dentata (Violaceae) used in South-Western Nigerian ethnomedicine and detection of cyclotides

Ethnopharmacological relevance People living in the tropical rain forest of South-Western Nigeria use Rinorea dentata (P. Beauv.) Kuntze (Violaceae) in ethno-veterinary medicine to facilitate parturition. There are no evidence-based pharmacological investigations for the uterotonic activity of this plant. Aims of study (i) Collection of data about the ethnopharmacological uses of R. dentata and evaluation of its uses and applications in health care; (ii) determining potential uterotonic effects in vitro, and (iii) chemical characterization of R. dentata, which is a member of the Violaceae family known to express circular cystine-knot peptides, called cyclotides. Materials and methods The ethnopharmacological use of R. dentata in settlement camps within the area J4 of Omo forest has been investigated by semi-structured questionnaires and open interviews. Use index analysis has been performed by seven quantitative statistical models. Respondents’ claim on the beneficial ethno-veterinary application of the plant to aid parturition has been investigated in vitro by myometrial contractility organ bath assays. The bioactive plant extract was screened by chemical derivatization and mass spectrometry-based peptidomics using reversed-phase HPLC fractionation and MALDI-TOF/TOF analysis. Results Based on the survey analysis, medicinal preparations of R. dentata have been used for anti-microbial and anti-malaria purpose in humans, and for aiding parturition in farm animals. The latter application was mentioned by one out of six respondents who claimed to use this plant for any medicinal purpose. The plant extract exhibited a weak uterotonic effect using organ bath studies. The plant contains cyclotides and the peptide riden A has been identified by de novo amino acid sequencing using mass spectrometry. Conclusion Few dwellers around the settlement camps of the tropical forest of Omo (Nigeria) use R. dentata for various health problems in traditional veterinary and human medicine. The weak uterotonic effect of the cyclotide-rich extract is in agreement with the low use value index obtained for this plant. Cyclotides have been reported in the genus Rinorea confirming the ubiquitous expression of these stable bioactive plant peptides within the family of Violaceae.