Fragmentation study of clerodane diterpenes from Casearia species by tandem mass spectrometry (quadrupole time-of-flight and ion trap).

RATIONALE: Clerodane-type diterpenes from Casearia species show important pharmacological activites such as antitumor, antimicrobial and anti-inflamatory. There are several mass spectrometry (MS)-based methods for identification of diterpenes; however, there is still a lack of MS procedures capable of providing characteristic fragmentation pathways for a rapid and unambiguous elucidation of casearin-like compounds. METHODS: Casearin-like compounds were investigated by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The fragmentation studies were carried out by tandem mass spectrometry in space (quadrupole time-of-flight (QTOF)) using different collision energies and also by tandem mass spectrometry in time (QIT) by selective isolation of product ions. RESULTS: Casearin-like compounds presented a predominance of sodium- and potassium-cationized precursor ions. Both QIT and QTOF techniques provided sequential neutral losses of esters related to the R1 to R5 substituents linked to the nucleus of the clerodane diterpenes. The fragmentation pathway is initiated with a cleavage of the ester moieties R2 followed by the elimination of the ester groups R3 , both losing neutral carboxylic acids. Using QIT, it was also possible to observe the cleavage of the ester groups R1 or R5 by MS4 experiments. CONCLUSIONS: Through a rational analysis of the fragmentation mechanisms of Casearia diterpenes it was possible to suggest an annotation strategy based on the sequential cleavages of the ester groups related to the R2 , R3 and R5 substituents. These results will assist studies of the dereplication and metabolomics involving casearin-like compounds present in complex extracts of Casearia species.

Climate effects on physicochemical composition of Syrah grapes at low and high altitude sites from tropical grown regions of Brazil.

The major areas of the world where viticulture is practiced enjoy temperate or cool temperature climates. When wine grapes are grown in tropical regions, edaphoclimatic factors result in distinct grape quality attributes, and production techniques also require significant adjustment. The objective of this study was to characterize the chemical compositions, in particular of phenolic compounds, of Syrah grapes grown in two location in northeast Brazil - these are also at widely different altitudes. A range of methods of phenolic extraction were used, along with classical chemical analyses including for organic acids, sugars, monomeric anthocyanins, flavonols, stilbene, condensed tannins and some of the monomeric and small oligomeric procyanidins. The regions and their diverse environments had a larger influence than harvest year. The grapes at higher altitude (Bahia, 1.100 m asl (metres above sea level) were characterized by higher levels of malic acid, anthocyanins and condensed tannins in the skins. The low-altitude grapes (Pernambuco, 350 m asl (metres above sea level) had higher levels of glucose, fructose, 3-O-acetylglucoside anthocyanins and condensed tannins in the seeds. Fruit composition was highly influenced by the region. In the low-altitude region, the grapes were characterized by higher tartaric and citric acid in the must, also of flavonols in skins and of tannins in the seeds. Meanwhile, the fruit from the high altitude, contained higher levels of malic and succinic acid in the must, and of anthocyanins and condensed tannins in the skins.

Chemical characteristics of grapes cv. Syrah (Vitis vinifera L.) grown in the tropical semiarid region of Brazil (Pernambuco state): influence of rootstock and harvest season.

BACKGROUND: Obtaining two or more successive annual vintages from the same vineyard is characteristic of regions with a tropical climate, such as the Submédio of the São Francisco Valley, Brazil. The present study aimed to characterize the chemical composition of grapes in four production cycles (i.e. two calendar years) when considering the interaction between cv. Syrah and two rootstocks. For a broad characterization, two methodologies for the extraction of phenolic compounds were used, as well as different methods of analysis. RESULTS: The results obtained showed that there was an influence of rootstock and harvest season. Grapes from vines grafted onto IAC 313 contained higher concentrations of total condensed tannins (skins) and flavanols than grapes from vines grafted onto 1103P. However, the grape samples from the vines grafted onto 1103P contained higher levels of monomeric anthocyanins than the grape samples from the vines grafted onto IAC 313. The first harvest season was characterized by higher concentrations of most phenolic compounds than the second harvest season. CONCLUSION: In the present study, it was possible to confirm that, in the semiarid region of Brazil, the interaction between the cultivar Syrah and the different rootstocks, as well as the climatic conditions in each harvest season, influenced the composition of the grapes, mainly in relation to phenolic compounds. © 2019 Society of Chemical Industry.

In vitro and in vivo anti-Helicobacter pylori activity of Casearia sylvestris leaf derivatives.

ETHNOPHARMACOLOGICAL RELEVANCE: The number of bacterial strains that are resistant to multiple conventional antimicrobial agents is increasing. In this context, natural products have been widely used as a strategy to treat diseases caused by bacteria. Infections by Helicobacter pylori have attracted attention because they are directly related to severe gastric medical conditions. Casearia sylvestris Swartz, popularly known as guaçatonga, is largely employed to treat gastric disorders in Brazilian folk medicine. This plant species has aroused much interest mainly because it displays anti-inflammatory activity and can act as an antiulcer agent. AIM OF THE STUDY: To evaluate the in vitro and in vivo anti-H. pylori action of C. sylvestris leaf derivatives incorporated or not in a nanostructured drug delivery system. MATERIALS AND METHODS: The essential oil (obtained by hydrodistillation) and ethanolic extract (obtained by maceration) were obtained from C. sylvestris leaves. The ethanolic extract was submitted to fractionation through solid phase extraction and column chromatography, to yield the ethanolic fractions. Hydrolyzed casearin J was achieved by submitting isolated casearin J to acid hydrolysis. The derivatives were chemically characterized by nuclear magnetic resonance (NMR), gas chromatography (GC), and gas chromatography-mass spectrometry (GC-MS) analyses. A nanostructured lipid system was used as drug delivery system. To assess the in vitro antibacterial activity of C. sylvestris leaf essential oil, ethanolic extract, and derivatives, microdilution, biofilm, and time-kill assays were performed against H. pylori ATCC 43504. Finally, the in vivo action was investigated by employing male Wistar rats experimentally infected with H. pylori. RESULTS: Many C. sylvestris leaf derivatives presented significant in vitro activity against H. pylori. Among the derivatives, fraction 2 (F2) was the most effective. In vivo tests showed that both the ethanolic extract and F2 decreased the ulcerative lesion size, but only the ethanolic extract eradicated H. pylori from the gastric lesions. Incorporation of plant derivatives in nanostructured lipid system blunted the in vitro action, as demonstrated by the microdilution assay. However, this incorporation improved the ethanolic extract activity against biofilms. CONCLUSION: C. sylvestris leaf derivatives are effective against H. pylori both in vitro and in vivo. According to phytochemical analyses, these derivatives are rich in terpenoids, which could be related to the anti-H. pylori action. Synergism could also underlie C. sylvestris efficacy judging from the fact that the sub-fractions and isolated compounds had lower activity than the extract. Incorporation in a nanostructured lipid system did not improve the activity of the compounds in our in vivo protocol.

[Surgical treatment of scoliosis in spinal muscular atrophy]. / Tratamento cirúrgico da escoliose na amiotrofia espinhal progressiva.

OBJECTIVE: To describe the early and late postoperative data from SMA patients with surgical procedure. METHOD: Clinical data and radiographic imaging from 14 SMA patients with surgical treatment of scoliosis were reviewed, and all were reassessed clinically with new spinal radiographs and a questionnaire. The mean follow-up were 22 months. The mean preoperative Cobb angle was 78.4 . All patients presented pelvic obliquity (mean 25.5 ) and 11 had cifosis. The mean age at time of surgery was 12 years and 3 months. All patients were treated with posterior spinal fusion with Luque-Galveston instrumentation in 12 and Cotrel-Dubousset instrumentation in 2. RESULTS: The average curve correction at the immediate postoperative was 64.3% for scoliosis and 36.4% for cifosis, with the pelvic obliquity correction of the 70.9%. The complications were liquoric fistula and infection earlier in one case, and lately wire looseness of T1 in 2 patients. It was detected mean lost of the correction at the final assessment of the 0.26 of the scoliosis and the 1.28 of the pelvic obliquity. Relatives and the patients related good improvement regarding to esthetic aspects, posture balance, body care, as well as respiratory problems. CONCLUSION: The spinal fusion for scoliosis in SMA patients has a satisfactory impact for esthetic, quality of life and respiratory function, with minimal lost of corrected deformities and few complications.