Octahedral ruthenium and magnesium naringenin 5-alkoxide complexes: NMR analysis of diastereoisomers and in-vivo antibacterial activity against Xylella fastidiosa.

Although many copper-based antimicrobial compounds have been developed to control pathogenic bacteria and fungi in plants and applied for crop protection, there is evidence that several plant pathogens have developed resistance to copper-based antimicrobial compounds, including some Xanthomonas species. Xylella is a bacterial genus belonging to the Xanthomonas family; and X. fastidiosa, which is responsible for citrus variegated chlorosis (CVC) in sweet orange, may develop resistance to one or more copper-based antimicrobials. Because of the time required for the development and approval of new antimicrobials for commercial use, the discovery of novel bactericidal compounds is essential before the development of resistance to the antimicrobials currently in use becomes widespread. Here, we explored the antimicrobial potential of two newly synthesized antimicrobials complexes and one natural compound against X. fastidiosa. Several nuclear magnetic resonance (NMR) assays with high resolution and sensitivity were developed to identify new diastereoisomers in the context of octahedral ruthenium - [Ru(narin)(phen)2]PF6-and magnesium naringenin 5-alkoxide - [Mg(narin)(phen)2]OAc - complexes, obtained in the present work. The NMR assays proved to be powerful tools for the identification of isomers in metal complexes. Moreover, a protocol for the in-vivo determination of the effects of these complexes against X. fastidiosa was developed. The main trunks of X. fastidiosa infected plants were injected with the two complexes as well as with the limonoid azadirachtin using a syringe; the number of bacterial cells in the plants following treatment was estimated via real-time quantitative PCR (qPCR). Importantly, the administration of both complexes and of azadirachtin drastically reduced the number of X. fastidiosa cells in vivo.

Clinical Impact of a Comprehensive Medication Management Service in Primary Health Care.

OBJECTIVE: To evaluate the clinical impact of a comprehensive medication management (CMM) service in a Brazilian primary health-care setting. METHODS: A quasi-experimental study has been carried out between July 2014 and November 2016 with patients who received the service in the primary care setting of a Brazilian city (n = 1057). Factors associated with drug therapy problems (DTP) detection in the initial assessment were evaluated by performing univariate and multivariate analyzes. To evaluate the impact of the CMM service, a linear regression model was constructed from the difference between the initial and final values of the clinical and laboratory parameters adjusted by multiple variables. RESULTS: A total of 1642 DTPs was identified, the most prevalent one being "nonadherence" (31.9%) and the "need for additional drug therapy" (22.9%). The use of 5 or more medications and the presence of 3 or more diseases were positively associated with the identification of 3 or more DTPs during the initial assessment. Even after multiple adjustments, a statistically significant reduction has been observed in the values of glycated hemoglobin, systolic blood pressure, low-density cholesterol, and total cholesterol. CONCLUSION: The CMM service contributed to the resolution of DTP and showed positive clinical impact in primary health care in the studied setting.

Rapid differentiation of graft Citrus sinensis with and without Xylella fastidiosa infection by mass spectrometry.

RATIONALE: Xylella fastidiosa causes citrus variegated chlorosis (CVC) in sweet orange trees. A diagnostic method for detecting CVC before the symptoms appear, which would inform citrus producers in advance about when the plant should be removed from the orchard, is essential for reducing pesticide application costs. METHODS: Chemometrics was applied to high-performance liquid chromatography diode array detector (HPLC-DAD) data to evaluate the similarities and differences between the chromatographic profiles. A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry selected reaction monitoring (LC/APCI-MS-SRM) method was developed to identify the major compounds and to determine their amounts in all samples. RESULTS: We evaluated the effect of this bacterium on the variation in the chemical profile in citrus plants. The organs of C. sinensis grafted on C. limonia were analyzed. Chemometrics was applied to the obtained data, and two major groups were differentiated. Flavonoids were observed in one group (leaves) and coumarins in the second (roots), both at higher concentrations in the plants with CVC symptoms than in those without the symptoms and those in the negative control. The rootstocks also interfered in the metabolism of the scion. CONCLUSIONS: The developed LC/APCI-MS-SRM method for detecting CVC before the symptoms appear is simple and accurate. It is inexpensive, and many samples can be screened per hour using 1 mg of leaves. Knowledge of the influence of the rootstock on the chemical profile of the graft is limited. This study demonstrates the effect of the rootstock in synthesizing flavonoids and increasing its content in all parts of the graft.

Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 µg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 µg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 µg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms.

Parasitic infection of the appendix as a cause of acute appendicitis.

The association between parasitic infection of the appendix and acute appendicitis has been widely investigated. The aim of this retrospective study was to evaluate the prevalence of parasitic infection of the appendix in a tropical area at southeast Brazil and to assess its possible relation to acute appendicitis in surgically removed appendices. Of the 1,600 appendectomies performed during a 10-year period, 24 (1.5%) were found to have helminths within the appendix. Enterobius vermicularis was observed in 23 of the 24 specimens (95.8%), and Taenia sp. was detected in only one case. Sixteen patients (66.7%) were less than 10 years old; 15 patients were male and nine female; 21 patients were white, and three were nonwhites. Pathologic analysis disclosed acute neutrophilic inflammation in the appendix wall in 12 of the 24 specimens and lymphoid hyperplasia in 10 of the 24 appendices. Gangrenous appendicitis was diagnosed in three cases, and peritonitis was found in 11 of the 24 infected appendices. The results of the present study indicate that E. vermicularis is the commonest worm found in the appendix and that its presence can cause pathologic changes ranging from lymphoid hyperplasia to acute phlegmonous inflammation with life-threatening complications like gangrene and peritonitis.