Chemical Differentiation of Plant Latexes and Their Anti-herbivory Activity against Thrips Frankliniella occidentalis.

Despite the extensive studies on latex, some fundamental questions on their chemical specialization and the factors influencing this specialization have yet to be investigated. To address this issue, latexes and their bearing tissues from diverse species were profiled by 1HNMR and GC-MS. Additionally, the antiherbivory activity of these materials was tested against thrips (Frankliniella occidentalis Pergande, 1895). The multivariate data analysis showed a clear separation between latexes and leaves from the same species. Conversely, the chemical profiles of latexes from different species were highly similar, that is, they displayed much less metabolic species-specificity. These shared chemical profiles of latexes were reflected in their overall higher mortality index (80.4% ± 7.5) against thrips compared with their bearing tissues (55.5% ± 14.9). The metabolites correlated to the antiherbivory activity of latexes were triterpenoids and steroids. However, the activity could not be attributed to any single terpenoid. This discrepancy and the reduction of the latex activity after fractionation suggested a complementary effect of the compounds when in a mixture as represented by the latex. Additionally, aqueous fractions of several latexes were found to possess simple spectra, even with only 1 metabolite. These metabolites were determined to be organic acids that might be involved in the modulation of the rate of latex coagulation, potentially increasing the sealing and trapping effects of the latex.

Infraspecific Chemical Variability and Biological Activity of Casearia sylvestris from Different Brazilian Biomes.

Casearia sylvestris is an outstanding representative of the Casearia genus. This representability comes from its distinctive chemical profile and pharmacological properties. This species is widespread from North to South America, occurring in all Brazilian biomes. Based on their morphology, 2 varieties are recognized: C. sylvestris var. sylvestris and C. sylvestris var. lingua. Despite the existence of data about their chemical composition, a deeper understanding of the specialized metabolism correlation and variation in respect to environmental factors and its repercussion over their biological activities was still pending. In this study, an UHPLC-DAD-based metabolomics approach was employed for the investigation of the chemical variation of 12 C. sylvestris populations sampled across 4 Brazilian biomes and ecotones. The correlation between infraspecific chemical variability and the cytotoxic and antioxidant activities was achieved by multivariate data analysis. The analyses showed that C. sylvestris var. lingua prevailed at Cerrado areas, and it was correlated with lower cytotoxic activity and high level of glycosylated flavonoids. Among them, narcissin and isorhamnetin-3-O-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranoside showed good correlation with the antioxidant activity. Conversely, C. sylvestris var. sylvestris prevailed at the Atlantic Forest areas, and it was associated with high cytotoxic activity and high content of clerodane diterpenoids. Different casearins showed good correlation (R2 = 0.3 - 0.70) with the cytotoxic activity. These findings highlighted the great complexity among different C. sylvestris populations, their chemical profile, and the related biological activities. Consequently, it can certainly influence the medicinal properties, as well as the quality and efficacy, of C. sylvestris phytomedicines.

Metabolic Profiling of Saponin-Rich Ophiopogon japonicus Roots Based on 1H NMR and HPTLC Platforms.

Ideally, metabolomics should deal with all the metabolites that are found within cells and biological systems. The most common technologies for metabolomics include mass spectrometry, and in most cases, hyphenated to chromatographic separations (liquid chromatography- or gas chromatography-mass spectrometry) and nuclear magnetic resonance spectroscopy. However, limitations such as low sensitivity and highly congested spectra in nuclear magnetic resonance spectroscopy and relatively low signal reproducibility in mass spectrometry impede the progression of these techniques from being universal metabolomics tools. These disadvantages are more notorious in studies of certain plant secondary metabolites, such as saponins, which are difficult to analyse, but have a great biological importance in organisms. In this study, high-performance thin-layer chromatography was used as a supplementary tool for metabolomics. A method consisting of coupling 1H nuclear magnetic resonance spectroscopy and high-performance thin-layer chromatography was applied to distinguish between Ophiopogon japonicus roots that were collected from two growth locations and were of different ages. The results allowed the root samples from the two growth locations to be clearly distinguished. The difficulties encountered in the identification of the marker compounds by 1H nuclear magnetic resonance spectroscopy was overcome using high-performance thin-layer chromatography to separate and isolate the compounds. The saponins, ophiojaponin C or ophiopogonin D, were found to be marker metabolites in the root samples and proved to be greatly influenced by plant growth location, but barely by age variation. The procedure used in this study is fully described with the purpose of making a valuable contribution to the quality control of saponin-rich herbal drugs using high-performance thin-layer chromatography as a supplementary analytical tool for metabolomics research.

Plant Latex, from Ecological Interests to Bioactive Chemical Resources.

Historically, latex-bearing plants have been regarded as important medicinal resources in many countries due to their characteristic latex ingredients. They have also often been endowed with a social or cultural significance in religious or cult rituals or for hunting. Initial chemical studies focused on the protein or peptide content but recently the interest extended to smaller molecules. Latex has been found to contain a broad range of specialized metabolites such as terpenoids, cardenolides, alkaloids, and phenolics, which are partly responsible for their antibacterial, antifungal, anthelmintic, cytotoxic, and insect-repellent activities. The diversity in biology and chemistry of latexes is supposedly associated to their ecological roles in interactions with exogenous factors. Latexes contain unique compounds that are different to those found in their bearing plants. Exploring the feasibility of plant latex as a new type of bioactive chemical resource, this review paper covers the chemical characterization of plant latexes, extending this to various other plant exudates. Also, the factors influencing this chemical differentiation and the production, transportation, and chemistry of the latex exudates are described, based on ecological and biochemical mechanisms. We also proposed a latex coagulation model involving 4 general conserved steps. Therefore, the inherent defensive origin of latexes is recognized as their most valuable character and encourages one to pay attention to these materials as alternative sources to discover metabolites with insecticidal or antimicrobial activity.

Coagulation Abnormalities Following NexoBrid Use: A Case Report.

Patients with major burn injury undergo a series of pathophysiologic changes that begin with a systemic inflammatory response and coagulation abnormalities, similar to those experienced by patients with sepsis or severe trauma. Coagulation changes in patients with burns are generally characterized by procoagulant abnormalities, but alterations in fibrinolysis and anticoagulation factors have also been observed. Around 40% of patients with major burn show changes on standard coagulation tests, and these have been related to the severity of the lesions, smoke inhalation, and administration of intensive fluid resuscitation therapy. Current surgical techniques for debridement of burn lesions are aggressive and associated with considerable blood loss. A fast-acting selective enzymatic debriding agent based on bromelain has been recently developed. NexoBrid is indicated for removing eschar in adults with deep partial- and full-thickness thermal burns. A potential effect of oral bromelain on hemostasis has been described, but it is uncertain whether NexoBrid application has a clinically relevant impact in this regard. We present the clinical case of a patient with burns who showed a coagulation abnormality shortly after NexoBrid use.

Broad range chemical profiling of natural deep eutectic solvent extracts using a high performance thin layer chromatography-based method.

Natural deep eutectic solvents (NADES) made mainly with abundant primary metabolites are being increasingly applied in green chemistry. The advantages of NADES as green solvents have led to their use in novel green products for the food, cosmetics and pharma markets. However, one of the main difficulties encountered in the development of novel products and their quality control arises from their low vapour pressure and high viscosity. These features create the need for the development of new analytical methods suited to this type of sample. In this study, such a method was developed and applied to analyse the efficiency of a diverse set of NADES for the extraction of compounds of interest from two model plants, Ginkgo biloba and Panax ginseng. The method uses high-performance thin-layer chromatography (HPTLC) coupled with multivariate data analysis (MVDA). It was successfully applied to the comparative quali- and quantitative analysis of very chemically diverse metabolites (e.g., phenolics, terpenoids, phenolic acids and saponins) that are present in the extracts obtained from the plants using six different NADES. The composition of each NADES was a combination of two or three compounds mixed in defined molar ratios; malic acid-choline chloride (1:1), malic acid-glucose (1:1), choline chloride-glucose (5:2), malic acid-proline (1:1), glucose-fructose-sucrose (1:1:1) and glycerol-proline-sucrose (9:4:1). Of these mixtures, malic acid-choline chloride (1:1) and glycerol-proline-sucrose (1:1:1) for G. biloba leaves, and malic acid-choline chloride (1:1) and malic acid-glucose (1:1) for P. ginseng leaves and stems showed the highest yields of the target compounds. Interestingly, none of the NADES extracted ginkgolic acids as much as the conventional organic solvents. As these compounds are considered to be toxic, the fact that these NADES produce virtually ginkgolic acid-free extracts is extremely useful. The effect of adding different volumes of water to the most efficient NADES was also evaluated and the results revealed that there is a great influence exerted by the water content, with maximum yields of ginkgolides, phenolics and ginsenosides being obtained with approximately 20% water (w/w).

Investigation of species and environmental effects on rhubarb roots metabolome using 1H NMR combined with high performance thin layer chromatography.

INTRODUCTION: The pharmacological activities of medicinal plants are reported to be due to a wide range of metabolites, therein, the concentrations of which are greatly affected by many genetic and/or environmental factors. In this context, a metabolomics approach has been applied to reveal these relationships. The investigation of such complex networks that involve the correlation between multiple biotic and abiotic factors and the metabolome, requires the input of information acquired by more than one analytical platform. Thus, development of new metabolomics techniques or hyphenations is continuously needed. OBJECTIVES: Feasibility of high performance thin-layer chromatography (HPTLC) were investigated as a supplementary tool for medicinal plants metabolomics supporting 1H nuclear magnetic resonance (1H NMR) spectroscopy. METHOD: The overall metabolic difference of plant material collected from two species (Rheum palmatum and Rheum tanguticum) in different geographical locations and altitudes were analyzed by 1H NMR- and HPTLC-based metabolic profiling. Both NMR and HPTLC data were submitted to multivariate data analysis including principal component analysis and orthogonal partial least square analysis. RESULTS: The NMR and HPTLC profiles showed that while chemical variations of rhubarb are in some degree affected by all the factors tested in this study, the most influential factor was altitude of growth. The metabolites responsible for altitude differentiation were chrysophanol, emodin and sennoside A, whereas aloe emodin, catechin, and rhein were the key species-specific markers. CONCLUSION: These results demonstrated the potential of HTPLC as a supporting tool for metabolomics due to its high profiling capacity of targeted metabolic groups and preparative capability.

Immunomodulatory Effect of Vitamin D after Allogeneic Stem Cell Transplantation: Results of a Prospective Multicenter Clinical Trial.

PURPOSE: We describe the results of a prospective multicenter phase I/II trial evaluating the impact of the use of vitamin D (VitD) from day -5 to +100 on the outcome of patients undergoing allogeneic transplantation (EudraCT: 2010-023279-25; ClinicalTrials.gov: NCT02600988). EXPERIMENTAL DESIGN: A total of 150 patients were included in three consecutive cohorts of 50 patients each group: control group (CG, not receive VitD); low-dose group (LdD, received 1,000 IU VitD daily); and high-dose group (HdD, 5,000 IU VitD daily). We measured levels of VitD, cytokines, and immune subpopulations after transplantation. RESULTS: No significant differences were observed in terms of cumulative incidence of overall and grades 2-4 acute GVHD in terms of relapse, nonrelapse mortality, and overall survival. However, a significantly lower cumulative incidence of both overall and moderate plus severe chronic GVHD (cGVHD) at 1 year was observed in LdD (37.5% and 19.5%, respectively) and HdD (42.4% and 27%, respectively) as compared with CG (67.5% and 44.7%, respectively; P < 0.05). In multivariable analysis, treatment with VitD significantly decreased the risk of both overall (for LdD: HR = 0.31, P = 0.002; for HdD: HR = 0.36, P = 0.006) and moderate plus severe cGVHD (for LdD: HR = 0.22, P = 0.001; for HdD: HR = 0.33, P = 0.01). VitD modified the immune response, decreasing the number of B cells and naïve CD8 T cells, with a lower expression of CD40L. CONCLUSIONS: This is the first prospective trial that analyzes the effect of VitD postransplant. We observed a significantly lower incidence of cGVHD among patients receiving VitD. Interestingly, VitD modified the immune response after allo-SCT. Clin Cancer Res; 22(23); 5673-81. ©2016 AACR.