Expression analysis of miRNAs and their putative target genes confirm a preponderant role of transcription factors in the early response of oil palm plants to salinity stress.

BACKGROUND: Several mechanisms regulating gene expression contribute to restore and reestablish cellular homeostasis so that plants can adapt and survive in adverse situations. MicroRNAs (miRNAs) play roles important in the transcriptional and post-transcriptional regulation of gene expression, emerging as a regulatory molecule key in the responses to plant stress, such as cold, heat, drought, and salt. This work is a comprehensive and large-scale miRNA analysis performed to characterize the miRNA population present in oil palm (Elaeis guineensis Jacq.) exposed to a high level of salt stress, to identify miRNA-putative target genes in the oil palm genome, and to perform an in silico comparison of the expression profile of the miRNAs and their putative target genes. RESULTS: A group of 79 miRNAs was found in oil palm, been 52 known miRNAs and 27 new ones. The known miRNAs found belonged to 28 families. Those miRNAs led to 229 distinct miRNA-putative target genes identified in the genome of oil palm. miRNAs and putative target genes differentially expressed under salinity stress were then selected for functional annotation analysis. The regulation of transcription, DNA-templated, and the oxidation-reduction process were the biological processes with the highest number of hits to the putative target genes, while protein binding and DNA binding were the molecular functions with the highest number of hits. Finally, the nucleus was the cellular component with the highest number of hits. The functional annotation of the putative target genes differentially expressed under salinity stress showed several ones coding for transcription factors which have already proven able to result in tolerance to salinity stress by overexpression or knockout in other plant species. CONCLUSIONS: Our findings provide new insights into the early response of young oil palm plants to salinity stress and confirm an expected preponderant role of transcription factors - such as NF-YA3, HOX32, and GRF1 - in this response. Besides, it points out potential salt-responsive miRNAs and miRNA-putative target genes that one can utilize to develop oil palm plants tolerant to salinity stress.

Chemical composition, anti-inflammatory and antinociceptive effects of the butanolic fraction of Annona nutans (Annonaceae) leaves.

The species Annona nutans (R. E. Fries) is a plant found in Bolivia, Paraguay, Argentina and the Brazilian cerrado. Considering the anti-inflammatory and antinociceptive activities of the hydrometanolic fraction (FHMeOH) of A. nutans leaves previously reported, the present study aimed to evaluate in vivo anti-inflammatory and antinociceptive activities of a subfraction obtained from FHMeOH, the butanolic fraction (FBuOHf). Intraperitoneal (i.p.) treatment with FBuOHf (50 and 100 mg · kg-1) inhibited paw edema induced by carrageenan. Moreover, FBuOHf (100 mg · kg-1, i.p.) also suppressed polymorphonuclear (PMN) leukocyte migration in the footpad. Regarding the antinociceptive activity, FBuOHf (50, 100, and 200 mg · kg-1, i.p.) inhibited acetic acid-induced abdominal writhing. In the formalin test, this fraction (200 mg · kg-1, i.p.) reduced licking time only in the inflammatory phase. The FBuOHf contents flavonoids and cinnamic acid derivatives, such as quercetin-3-O-galactoside, quercetin-3-O-glucoside, isorhamnetin-3-O-galactoside, quercetin-3-O-ß-D-apio-furanosyl-(1→2)-galactopyranoside and chlorogenic acid, identified and quantified by LC-MS. The FBuOHf possesses anti-inflammatory and peripheral antinociceptive activities.

Anti-inflammatory and antinociceptive activities of a phenylpropanoid-enriched fraction of Duguetia furfuracea.

A previous study reported the in vivo anti-inflammatory and antinociceptive activities of essential oil of the underground stem bark of Duguetia furfuracea, termed EODf. This study aimed to obtain a phenylpropanoid-enriched fraction from the D. furfuracea (EFDf) essential oil and to investigate its anti-inflammatory and antinociceptive effects. The chemical composition of the EFDf was determined by gas chromatography-mass spectrometry (GC-MS). The in vivo anti-inflammatory activity was evaluated with a lipopolysaccharide (LPS)-induced paw oedema model. The effects of the EFDf on the polymorphonuclear leukocyte recruitment and the inducible nitric oxide synthase (iNOS) expression were evaluated in mice footpads. Moreover, the in vivo antinociceptive effect was assayed using the formalin test and the LPS-induced thermal hyperalgesia model. In the EFDf, 8 major compounds were identified, with α-asarone (36.4%) and 2,4,5-trimethoxystyrene (27.8%) the main constituents. A higher concentration of phenylpropanoid derivatives was found in the EFDf, 64.2% compared to the EODf (38%). The oral (p.o.) treatment with the EFDf at a dose of 3 mg/kg significantly attenuated the paw oedema, polymorphonuclear leukocyte migration, iNOS expression, and tumour necrosis factor alpha (TNF-α) production. The EFDf (10 and 30 mg/kg) also inhibited both phases of the formalin test and caused a significant increase in the reaction time in the LPS-induced thermal hyperalgesia model. Finally, EFDf-treated animals did not show any alteration of motor coordination. The results suggest that the enrichment of 2,4,5-trimethoxystyrene and α-asarone enhances the anti-inflammatory activity of the EFDf compared to the EODf. In contrast, the antinociception promoted by the EFDf was similar to the EODf and was mediated via activation of adenosinergic and opioidergic receptors.

Anti-inflammatory and central and peripheral anti-nociceptive activities of α-asarone through the inhibition of TNF-α production, leukocyte recruitment and iNOS expression, and participation of the adenosinergic and opioidergic systems.

Alpha-asarone has been found to possess many pharmacological activities, which can improve cognitive function and exert anti-oxidant, anxiolytic, anti-epileptic and protective effects against endothelial cell injury. The anti-inflammatory activity of α-asarone was evaluated using lipopolysaccharide (LPS)-induced paw oedema. Moreover, leukocyte migration, inducible nitric oxide synthase (iNOS) expression and tumour necrosis factor-alpha (TNF-α) levels were quantified in footpads. Formalin and LPS-induced thermal hyperalgesia models were generated using adenosinergic, opioidergic, serotonergic and muscarinic receptor antagonists. The effects on motor coordination were evaluated by means of the rota-rod test. Oral treatment (p.o.) with α-asarone (3 mg/kg) significantly inhibited paw oedema by 62.12 and 72.22%, 2 and 4 h post LPS injection, respectively. Alpha-asarone (3 mg/kg, p.o.) attenuated the inflammatory infiltrate 1, 3 and 6 h after LPS injection. Furthermore, α-asarone (3 mg/kg, p.o.) suppressed iNOS expression and TNF-α production, 6 and 1 h after inflammatory stimulus, respectively. Alpha-asarone (3, 10 and 30 mg/kg, p.o.) inhibited both phases of formalin-induced licking. In the hot-plate test, α-asarone (10 and 30 mg/kg, p.o.) increased the latency to response 3 and 5 h post LPS stimulus. Caffeine and naloxone abolished the central anti-nociceptive effect of α-asarone (neurogenic phase of formalin and hot plate tests), suggesting the participation of the adenosinergic and opioidergic systems. Furthermore, naloxone reversed the peripheral activity of α-asarone (inflammatory phase of formalin test), indicating the possible involvement of the opioidergic pathway. In the rota-rod test, α-asarone did not change motor coordination. These findings suggest that α-asarone has anti-inflammatory, peripheral and central anti-nociceptive effects and could represent a promising agent for future research.

Are the Statins promising antifungal agents against invasive candidiasis?

The medical importance of intra-abdominal candidiasis (IAC) contrasts with the limited number of pharmacological treatment options available and the increasing rate of resistance to antifungal drugs. Thus, the repositioning of compounds in clinical use can contribute to the broadening of treatment possibilities for this infection. Statins, a class of drugs used to reduce cardiovascular event risk, have shown antiparasitic, antibacterial, and antiviral activities; however, their antifungal effects remain poorly studied. In this context, the present study aimed to elucidate the antifungal potential of six statins in vitro, as well as to evaluate the therapeutic use of fluvastatin in a mouse model of IAC. The biological effects of statins were evaluated against Candida spp., through the determination of the minimum inhibitory concentration (MIC). For the statins that showed activity, the fungicidal concentration, toxicity/selectivity, synergism with azoles and polyenes, phenotypic effects, and activity against virulence factors were also determined. Atorvastatin, rosuvastatin and fluvastatin were highly active, especially against C. albicans (MIC < 1-128 µg.mL-1) and C. glabrata (MIC 32-64 µg.mL-1). Fluvastatin and atorvastatin were selective for C. albicans in baby hamster kidney (BHK) cells. Moreover, all active statins in the antifungal assay showed high selectivity for fungal cells over bacteria. The combination of atorvastatin, rosuvastatin, and fluvastatin with azoles was associated with a synergistic effect. Active statins do not act on the fungal membrane or wall, but instead stimulate farnesol-dependent pathogenicity factors such as yeast-to-hyphal transition and biofilm generation. Fluvastatin treatment was evaluated in a mouse model of IAC, showing stimulation of the extra-hepatic dissemination of C. albicans but improvements in renal, splenic, and hepatic histological aspects. In conclusion, statins have potent antifungal activity in vitro, but the therapeutic effect in vivo is restricted to their anti-inflammatory activity.

Chemical composition and evaluation of the anti-inflammatory and antinociceptive activities of Duguetia furfuracea essential oil: Effect on edema, leukocyte recruitment, tumor necrosis factor alpha production, iNOS expression, and adenosinergic and opioidergic systems.

ETHNOPHARMACOLOGICAL RELEVANCE: Duguetia furfuracea (A. St. -Hil.) Saff. (Annonaceae) is commonly known in Brazil as "araticum-seco," and its root is used in folk medicine to treat inflammatory and painful disorders. However, no studies have been performed to evaluate these therapeutic activities. AIM OF THE STUDY: Investigate the chemical composition, anti-inflammatory and antinociceptive effects, and elucidate the possible antinociceptive mechanisms of action from the essential oil of D. furfuracea (EODf) underground stem bark. MATERIALS AND METHODS: Chemical composition was determined by gas chromatography and mass spectrometry (GC/MS). The paw edema induced by LPS, formalin-induced nociception, LPS-induced thermal hyperalgesia and rota-rod tests in vivo were used to evaluate the anti-inflammatory and antinociceptive effects in addition to the alteration on motor coordination. Histological analyses and an immunohistochemistry assay for iNOS were performed on mouse footpads of naive, control, 10 mg/kg EODf, and 10 mg/kg indomethacin (Ind) groups. The samples were removed at 1, 3, and 6 h after subplantar injection of LPS. In addition, the involvement of the adenosinergic, opioidergic, serotonergic, and cholinergic systems were investigated, in order to elucidate possible antinociceptive mechanisms. RESULTS: Twenty-four volatile constituents were detected and identified. (E)-asarone (21.9%), bicyclogermacrene (16.7%), 2,4,5-trimethoxystyrene (16.1%), α-gurjunene (15%), cyperene (7.8%), and (E)-caryophyllene (4.6%) were major compounds found in EODf. Oral treatment (p.o.) with EODf (1, 3, and 10 mg/kg) significantly inhibited the paw edema induced by LPS. At 10 mg/kg EODf promoted inhibition of tumor necrosis factor alpha (TNF-α) production, recruitment of polymorphonuclear (PMN) leukocytes and inducible nitric oxide synthase (iNOS) expression in paw tissue. EODf (10 and 30 mg/kg, p.o.) also reduced licking time in both phases of the formalin test and it had a significant effect on the LPS-induced thermal hyperalgesia model. The administration of caffeine (Caf) and naloxone (Nal) reversed the antinociceptive activity of EODf, in the first phase of the formalin test and in the LPS-induced thermal hyperalgesia model. Moreover, Nal was also able to abolish the antinociception caused by EODf, in the second phase of formalin test. In the rota-rod test, EODf-treated animals did not show any alteration of motor coordination. CONCLUSIONS: Our findings indicate that EODf underground stem bark produces anti-inflammatory and both central and peripheral antinociceptive effects. Furthermore, the antinociceptive activity of EODf underground stem bark is possibly mediated by adenosinergic and opioidergic pathways, and its properties do not induce effects on motor coordination. These results support the use of the folk medicine, D. furfuracea root, to treat inflammation and painful conditions.

Multielement analysis of plant extracts with potential use in the treatment of peptic ulcers by synchrotron radiation total reflection X-ray fluorescence.

Some plants popularly employed for the treatment of peptic ulcers have proved to be attractive sources of new drugs. Despite extensive research, the pharmacological and toxicological potentials of these plants are not fully understood. In this context, the aim of this work was to analyze the multielemental composition of the methanolic extracts of three of those plants, Alchornea glandulosa (AG), Davilla elliptica (DE) and Davilla nitida (DN), with the intention of contributing to the understanding of the mechanisms of action of these extracts. For this purpose, we used the analytical technique of total reflection X-ray fluorescence (TXRF) by synchrotron radiation at the Brazilian Synchrotron Light Source (LNLS/CNPEM). It was possible to determine the concentrations of the elements: P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Rb and Br in all of the samples. Selenium (Se) was detected only in the DN extract. An inverse relationship between the concentrations of elements with proven effectiveness and the gastroprotective activity of extracts considering induction protocols with ethanol and non-steroidal anti-inflammatory drugs (NSAIDs) was obtained. This data suggests that the function of the extract is not only associated with providing the elements for restoring the gastric mucosa but that it also promotes the displacement of these elements from other parts of the mucosa to the damaged area. Correlations between the concentrations of the elements were also obtained. In the DE extract, which is the most effective extract for both induction protocols, the obtained correlations were above 70% among almost all of the elements, and no anticorrelations were found. For the other two extracts, in the less effective extract (AG) anticorrelations above 70% were predominantly found. Meanwhile, in the DN extract, a few high anticorrelations were found, which may explain its intermediate stage of effectiveness.

Vitamin D intoxication through errors in administration: a case report / Intoxicação de vitamina D por erro de administração: relato de caso

Abstract Vitamin D intoxication caused by the irregular consumption of medications is a major concern in geriatric health. Due to errors in administering such vitamins and medical malpractice, many patients lack the proper management of vitamin supplementation, considering what is actually prescribed. The present study, which aims to report on intoxication by this vitamin, describes an elderly couple who lived alone and divided their household tasks. The wife, who is the main focus of the report, was lucid but suffered musculoskeletal disorders and used a wheelchair, while the husband could function physically. The wife was hospitalized with a clinical profile of delirium. Intoxication is a major cause of metabolic encephalopathy, which explains how the case developed. The co-adjuvant was the husband, who suffered apparent mild cognitive impairment, and modified the doctor's dosage of vitamin D alone, contradicting the guidance of the family. The family monitored the health status of the couple through weekly telephone calls to check if their medications were being taken properly. After investigation with new anamneses and a review of medical records, intoxication was confirmed due to an error in the amount of the drug administered over a prolonged period. It is extremely important to be aware of the clinical profile of hypercalcemia and how to treat the same. In geriatrics, diagnosis should involve both clinical treatment and special care to understand the daily routine of elderly persons in order to avoid further repercussions. AU

Resumo A intoxicação por vitamina D merece ser destacada em geriatria devido ao consumo irregular de medicamentos. Muitos pacientes não têm manejo competente da suplementação vitamínica comparada a real prescrição, tanto na administração quanto por negligência médica. Este artigo, que tem como objetivo relatar sobre a intoxicação por essa vitamina, mostra o cenário de um casal de idosos que moravam sozinhos, dividindo as tarefas domiciliares, sendo que a esposa tinha uma mente lúcida e o marido um corpo funcionante, para que se estabelecesse a vivência. A esposa, alvo principal do relato, possuía distúrbios osteomusculares em condição de cadeirante, assim a vitamina D se fazia necessária, e se apresentou na internação com o quadro clínico de delirium. Destaca-se que essa intoxicação é importante causa de encefalopatia metabólica, o que explica o seguimento do caso. O coadjuvante foi o marido, com aparente distúrbio cognitivo leve, que modificou por conta própria a posologia da vitamina D indicada pelo médico, apesar da orientação da família. A família fazia controle da situação da saúde do casal através de ligações telefônicas semanais, conferindo se as medicações estavam sendo apropriadamente tomadas. Após investigação com novas anamneses e revisão de prontuário, ficou confirmada a intoxicação devido a um erro de quantidade de administração da droga por tempo prolongado. É de extrema importância que se tenha conhecimento sobre o quadro clínico da hipercalcemia e como manuseá-lo. Em geriatria, o diagnóstico deve ter a solução clínica somada a um cuidado especial de entender o que ocorre no cotidiano da pessoa idosa para que assim se evite novas repercussões. AU

Metabolic fingerprinting analysis of oil palm reveals a set of differentially expressed metabolites in fatal yellowing symptomatic and non-symptomatic plants.

INTRODUCTION: Oil palm (E. guineensis), the most consumed vegetable oil in the world, is affected by fatal yellowing (FY), a condition that can lead to the plant's death. Although studies have been performed since the 1980s, including investigations of biotic and abiotic factors, FY's cause remains unknown and efforts in researches are still necessary. OBJECTIVES: This work aims to investigate the metabolic expression in plants affected by FY using an untargeted metabolomics approach. METHOD: Metabolic fingerprinting analysis of oil palm leaves was performed using ultra high liquid chromatography-electrospray ionization-mass spectrometry (UHPLC-ESI-MS). Chemometric analysis, using principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA), was applied to data analysis. Metabolites identification was performed by high resolution mass spectrometry (HRMS), MS/MS experiments and comparison with databases and literature. RESULTS: Metabolomics analysis based on MS detected more than 50 metabolites in oil palm leaf samples. PCA and PLS-DS analysis provided group segregation and classification of symptomatic and non-symptomatic FY samples, with a great external validation of the results. Nine differentially expressed metabolites were identified as glycerophosphorylcholine, arginine, asparagine, apigenin 6,8-di-C-hexose, tyramine, chlorophyllide, 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine, proline and malvidin 3-glucoside-5-(6″-malonylglucoside). Metabolic pathways and biological importance of those metabolites were assigned. CONCLUSION: Nine metabolites were detected in a higher concentration in non-symptomatic FY plants. Seven are related to stress factors i.e. plant defense and nutrient absorption, which can be affected by the metabolic depression of these compounds. Two of those metabolites (glycerophosphorylcholine and 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine) are presented as potential biomarkers, since they have no known direct relation to plant stress.

Growth curve, biochemical profile and phytochemical analyses in calli obtained from the procambium segments of Bacupari

Garcinia brasiliensis, popularly known as Bacupari, is native to the Amazon and commonly used in folk medicine for its therapeutic properties. This plant is rich in bioactive compounds like benzophenones. However, there are no works about the in vitro establishment and achievement of secondary metabolites in this plant. Thus, the aim of this work was to determine the growth curve and to perform the biochemical and phytochemical analyses in calli obtained from the procambium segments of Bacupari. The growth curve of calli followed a sigmoidal pattern, with four distinct phases (lag, exponential, linear, deceleration). Total soluble sugars were higher on the inoculation day and the reducing sugars on the 20 th day. Amino acids increased from the 60 th day up to the stabilization on the 120 th day. The protein content varied, but it seemed to be related to the amino acids metabolism. The phytochemical screening showed the presence of phenolic and flavonoid compounds in the calli and the HPLC analysis allowed the identification of Fukugetin, Guttiferone A and 7-epiclusianone.

Avaliação citotóxica em três linhagens de células tumorais das frações obtidas da casca do caule de Salacia crassifolia (MART. ex. Schult.) G. Dom. (Celastraceae) / Assessment of cytotoxicity in three tumor cell lines of the fractions attained from the stem bark of Salacia crassifolia (Mart. ex. Schult.) G. Dom (Celastraceae)

A utilização de plantas pela humanidade com fins medicinais é uma das formas mais antigas para tratamento, cura e prevenção de doenças, dentre elas o câncer, cujas espécies vegetais são encontradas em elevada diversidade no ecossistema. Com essa perspectiva, se optou por um arbusto típico do Cerrado, Salacia crassifolia (Celastraceae), para um screening inicial na determinação do potencial citotóxico desta espécie in vitro. Na medicina popular, S. crassifolia é utilizada no tratamento de problemas renais, tosse crônica, dores de cabeça, cicatrizantes, ulcerogênicas e na terapia da malaria. As frações: hidroalcoólica (SCCcM-W), diclorometânica(SCCcM-D), hexânica (SCCcM-H) e acetato de etila (SCCcM-A) da casca do caule foram submetidas a testes de citotoxicidade in vitro frente às linhagens MDA-MB-435(melanoma), HCT-8 (cólon-humano) e SF-295 (sistema nervoso central) utilizando o método colorimétrico MTT. As frações hexânica (SCCcM-H) e de acetato de etila(SCCcM-A) foram aquelas que proporcionaram citotoxicidade significativas frente às células tumorais analisadas.

For millennia humans have been using plants for medicinal purposes to treat, cure, or prevent diseases, including cancer. Brazilian ecosystem has a high diversity of plant species. From this perspective, a species found in the Cerrado, Salacia crassifolia (Celastraceae), was selected, for an initial screening to determine the cytotoxic potential of this species "in vitro". S. crassifolia has been widely used in traditional medicine to treat kidney diseases, chronic cough, headaches, healing, ulcerogenic, and for treatment of malaria. The fractions: hydroalcoholic (SCCcM-W), dichloromethane(SCCcM-D), hexane (SCCcM-H) and ethyl acetate (SCCcM-A) from the stem bark were tested for cytotoxicity "in vitro" to within MDA -MB-435(melanoma), HCT-8 (human-colon), and SF-295 (CNS) using the MTT colorimetric method. The hexane (SCCcM-H) and ethyl acetate (SCCcM-A) fractions were those that provided significant cytotoxicity against analyzed tumor cells.