Antileukemic, Antioxidant, Anti-Inflammatory and Healing Activities Induced by a Polyphenol-Enriched Fraction Extracted from Leaves of Myrtus communis L.

Natural products have offered a number of exciting approaches in cancer treatment over the years. In this study, we investigated the prophylactic and therapeutic effects of the polyphenol-enriched fraction extracted from Myrtus communis (PEMC) on acute and chronic leukemia. According to the UHPLC-MSn, the fraction is rich in flavonoids. Protective activity of the PEMC was assessed by evaluating the antioxidant, anti-inflammatory, wound healing, and hemolysis potential in a series of in vivo and in vitro assays, while the therapeutic approach consisted of the evaluation of cytotoxic activity of the PEMC against HL60 and K562 leukemia cell lines. Safety of the fraction was also evaluated on a non-cancerous Vero cell line and by an acute toxicity test performed in mice. The PEMC demonstrated a significant anti-inflammatory and healing potential. The activities found at the dose of 100 mg/kg were better than those observed using a reference drug. The PEMC demonstrated a significant antioxidant effect and a specific cytotoxicity towards HL60 (IC50 = 19.87 µM) and K562 (IC50 = 29.64 µM) cell lines being non-toxic to the Vero cell line. No hemolytic activity was observed in vitro and no toxicity effect was found in mice. Thus, the PEMC has a pharmacological potential as both preventive and therapeutic agent. However, further research is necessary to propose its mechanism of action.

Effect of low-level laser therapy on the membrane induced by the Masquelet technique at an orthotopic site in rabbits.

PURPOSE: To evaluate the low-level laser therapy (LLLT) on the membrane induced by the Masquelet technique in rabbits. METHODS: Twelve Norfolk rabbits at approximately 3 months of age were used. A 1-cm segmental defect was induced in both radii, which were filled with polymethylmethacrylate cylinder. LLLT was used postoperatively in the bone defect of one of the forelimbs every 48 hours for 15 days. Six rabbits were euthanatized on third and sixth postoperative weeks. RESULTS: In both forelimbs, radiographs showed new bone growth from radius cut ends on the third postoperative week and more advanced stage on the sixth postoperative week. Ultrasound showed induced membrane one week after the surgery. Histologically, there were no significant differences in the semi-quantitative score of inflammation intensity, total number of blood vessels, bone metaplasia, and collagen. The average thicknesses were 2,050.17 and 1,451.96 µm for control membranes and 2,724.26 and 2,081.03 µm for irradiated membranes, respectively, on third and sixth postoperative weeks. Vascular endothelial growth factor A (VEGF-A) and platelet derived growth factor (PDGF) expression were present in the induced membranes of control and irradiated forelimbs, but there was no significant difference. CONCLUSIONS: Based on assessment methods, it was not possible to demonstrate the effect of LLLT on the induced membrane.

Potential of Piper spp. as a source of new compounds for the leishmaniases treatment.

Current treatment guidelines for leishmaniasis is based on chemotherapy with drugs that show a set of limitations such as high cost, toxicity, difficult route of administration, and lack of efficacy in endemic areas. In this context, phytopharmaceutical products and herbal medicines emerge as promising alternatives for developing new treatment against leishmaniasis. This review discusses the perspectives of leishmaniasis treatment based on natural products and phytotherapy highlighting the Piper genus, especially P. aduncun and P. mollicomum Kunth covering the period of 1998 to 2020. Leishmanicidal activity of pure compounds of Piper spp. [3-(3,4,5-trimethoxyphenyl) propanoic acid, 3-chlorosintenpyridone, 2'-hydroxy-3',4',6'-trimethoxy-chalcone, cardamonin, conocarpan, cubebin, eupomatenoid, flavokavain B, ( +)-(7R,8S)-epoxy-5,6-didehydrokavain, N-[7-(3',4'-methylenedioxypheny l-2(E),4(E)-heptadienoyl-pyrrolidine, N-[7-(3',4'-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl-pyrrolidine, piperovatine, pellitorine, and piplartine (piperlongumine)] were proved against the promastigote and amastigote forms of parasite related with cutaneous (L. (L.) amazonensis, L. (V.) braziliensis, and L. (V.) guyanensis) and visceral (L. (L.) donovani, L. (L.) chagasi, and L. (L.) infantum). We also discussed the perspective of leishmaniasis treatment, considering the potential synergism between different promising species of Piper, presenting some interesting interaction possibilities for future studies between plants. Finally, the necessary steps for technological development of phytomedicines and herbal medicines with the desirable quality requirements for medicines are highlighted. The data presented here highlight the use of Piper spp. as source of pharmacological compounds that can lead to effective, safe, and inexpensive treatments for leishmaniasis.

Axenic amastigotes of Leishmania species as a suitable model for in vitro studies.

Leishmania spp. are etiological agents of infection diseases, which in some cases can be fatal. The main forms of their biological cycle, promastigotes and amastigotes, can be maintained in vitro. While promastigotes are easier to maintain, amastigotes are more complex and can be obtained through different ways, including infection assays of tissues or in vitro cells, and differentiation from promastigotes to axenic amastigotes. Several protocols have been proposed for in vitro differentiation for at least 12 Leishmania spp. of both subgenera, Leishmania and Viannia. In this review we propose a critical summary of axenic amastigotes induction, as well as the impact of these strategies on metabolic pathways and regulatory networks analyzed by omics approaches. The parameters used by different research groups show considerable variations in temperature, pH and induction stages, as highlighted here for Leishmania (Viannia) braziliensis. Therefore, a consensus on strategies for inducing amastigogenesis is necessary to improve accuracy and even define stage-specific biomarkers. In fact, the axenic amastigote model has contributed to elucidate several aspects of the parasite cycle, however, since it does not reproduce the intracellular environment, its use requires several precautions. In addition, we present a discussion about using axenic amastigotes for drug screening, suggesting the need of a more sensitive methodology to verify cell viability in these tests. Collectively, this review explores the advantages and limitations found in studies with axenic amastigotes, done for more than 30 years, and discuss the gaps that impair their use as a suitable model for in vitro studies.

Effect of low-level laser therapy on the membrane induced by the Masquelet technique at an orthotopic site in rabbits

ABSTRACT Purpose: To evaluate the low-level laser therapy (LLLT) on the membrane induced by the Masquelet technique in rabbits. Methods: Twelve Norfolk rabbits at approximately 3 months of age were used. A 1-cm segmental defect was induced in both radii, which were filled with polymethylmethacrylate cylinder. LLLT was used postoperatively in the bone defect of one of the forelimbs every 48 hours for 15 days. Six rabbits were euthanatized on third and sixth postoperative weeks. Results: In both forelimbs, radiographs showed new bone growth from radius cut ends on the third postoperative week and more advanced stage on the sixth postoperative week. Ultrasound showed induced membrane one week after the surgery. Histologically, there were no significant differences in the semi-quantitative score of inflammation intensity, total number of blood vessels, bone metaplasia, and collagen. The average thicknesses were 2,050.17 and 1,451.96 μm for control membranes and 2,724.26 and 2,081.03 μm for irradiated membranes, respectively, on third and sixth postoperative weeks. Vascular endothelial growth factor A (VEGF-A) and platelet derived growth factor (PDGF) expression were present in the induced membranes of control and irradiated forelimbs, but there was no significant difference. Conclusions: Based on assessment methods, it was not possible to demonstrate the effect of LLLT on the induced membrane.

Local botanical knowledge of native food plants in the semiarid region of Brazil.

BACKGROUND: This study aimed to investigate the local botanical knowledge of native food plants in three rural communities, located in the semiarid region of Paraíba State, Brazil, verifying possibilities of differences of knowledge among communities and between men and women. METHODS: Semi-structured interviews about native plant knowledge and use were conducted with all householders in each community, totaling 117 informants. The species similarity among the communities of Pau D'Arco, Várzea Alegre, and Barroquinha was compared with Jaccard index, and the use value index (UVgeneral, UVcurrent, UVpotential) was used to determine the most important species. The Kruskal-Wallis test was used to compare the use values among communities and genders. The consensus factor among the informants was calculated according to the uses cited, and the Wilcoxon test was used to compare the use values between men and women. RESULTS: We recorded 9 species belonging to 8 genera and 8 families in Várzea Alegre; 10 species, 9 genera, and 9 families in Barroquinha; and 7 species, 7 genera and 7 families in Pau D'Arco. Spondias tuberosa Arruda (Anacardiaceae) in Várzea Alegre, Spondias sp. (Anacardiaceae) in Barroquinha, and Ximenia americana L. (Olacaceae) in Pau D'Arco were the most prominent species. Preparation methods are slightly different in the three communities, and there is low similarity about species use among the communities. Regarding gender, the analysis of use value among the communities evidenced significant differences only for UVgeneral among women, specifically between Barroquinha and Pau D'Arco. For men and women within each community, there is a difference only for UVpotential in Barroquinha. CONCLUSION: This study showed that the residents of the three rural communities have limited knowledge of native food plants found in their communities, but they know where to find them, which parts they may use and how to consume them. The fact is that men know plants that are more distant from the residences and women know those that are next to them.

Low-level laser irradiation induces a transcriptional myotube-like profile in C2C12 myoblasts.

Low-level laser irradiation (LLLI) has been used as a non-invasive method to improve muscular regeneration capability. However, the molecular mechanisms by which LLLI exerts these effects remain largely unknown. Here, we described global gene expression profiling analysis in C2C12 myoblasts after LLLI that identified 514 differentially expressed genes (DEG). Gene ontology and pathway analysis of the DEG revealed transcripts among categories related to cell cycle, ribosome biogenesis, response to stress, cell migration, and cell proliferation. We further intersected the DEG in C2C12 myoblasts after LLLI with publicly available transcriptomes data from myogenic differentiation studies (myoblasts vs myotube) to identify transcripts with potential effects on myogenesis. This analysis revealed 42 DEG between myoblasts and myotube that intersect with altered genes in myoblasts after LLLI. Next, we performed a hierarchical cluster analysis with this set of shared transcripts that showed that LLLI myoblasts have a myotube-like profile, clustering away from the myoblast profile. The myotube-like transcriptional profile of LLLI myoblasts was further confirmed globally considering all the transcripts detected in C2C12 myoblasts after LLLI, by bi-dimensional clustering with myotubes transcriptional profiles, and by the comparison with 154 gene sets derived from previous published in vitro omics data. In conclusion, we demonstrate for the first time that LLLI regulates a set of mRNAs that control myoblast proliferation and differentiation into myotubes. Importantly, this set of mRNAs revealed a myotube-like transcriptional profile in LLLI myoblasts and provide new insights to the understanding of the molecular mechanisms underlying the effects of LLLI on skeletal muscle cells.

Antileishmanial Activity of 2-Methoxy-4H-spiro-[naphthalene-1,2'-oxiran]-4-one (Epoxymethoxy-lawsone): A Promising New Drug Candidate for Leishmaniasis Treatment.

Epoxymethoxylawsone is a naphthoquinone derivative promising as drug candidate for the treatment of leishmaniases. In the present work the effectiveness of epoxymethoxylawsone, and meglumine antimoniate on Leishmania (Leishmania) amazonensis parasites and on mice paw lesions of infected BALB/c mice was assessed. In an intracellular amastigotes assay, the half-maximal inhibitory concentration (IC50) value for epoxymethoxylawsone was slightly higher (1.7-fold) than that found for meglumine antimoniate. The efficacy of both drugs became more evident after 48 h of exposure when either the oxirane compound and reference drug reached 18-fold and 7.4-fold lower IC50 values (0.40 ± 0.001 µM and 0.60 ± 0.02 µM), respectively. Promastigotes were also affected by epoxymethoxylawsone after 24 h of incubation (IC50 = 45.45 ± 5.0 µM), but with IC50 6-fold higher than those found for intracellular amastigotes. Cytotoxicity analysis revealed that epoxymethoxylawsone (CC50 = 40.05 ± µM) has 1.7-fold higher effects than meglumine antimoniate (CC50 = 24.14 ± 2.6 µM). Treatment of the paw lesion in infected BALB/c mice with epoxymethoxy-lawsone led to a significant 27% reduction (p < 0.05) of the lesion size, for all administrated doses, compared to the control group. Lesion reduction was also detected after mice treatment with meglumine antimoniate, reaching 31.0% (0.23 mg of Sb(V)/Kg/day and 2.27 mg of Sb(V)/Kg/day) and 64.0% (22.7 mg of Sb(V)/Kg/day). In addition, mice lesion ultrastructural changes were evidenced in amastigotes. The set of data gathered here indicate that epoxymethoxylawsone has pronounced effects on parasites and merits furthering to the preclinical stage.

Analysis of spectra from portable handheld gamma-ray spectrometry for terrain comparative assessment.

Geological characteristics can have impacts on societal development by, e.g., geotechnical issues and radiological hazard levels. Due to urban sprawl, there is an increasing need for detailed geological assessment. In this work are analysed data from portable handheld gamma-ray spectra (K, eU and eTh) obtained in granitic and Silurian metaclastic outcrops as well as in an profile, roughly N-S, on soil covered terrains transecting a mapped contact between these rock types (the profile's northern extremity is at locations mapped as granite). Estimations from gamma-ray spectra were studied by univariate and multivariate analyses. K, eU and eTh values were higher on granite in relation to Silurian metaclastic rocks. The northern extremity of the profile showed clearly higher contents of eTh and this contrast was supported by univariate statistical tools (normality plot and Wilk-Shapiro test; boxplots). A ternary plot with the contribution of the elements to gamma-ray absorbed dose showed the separation of granite from Silurian metaclastic rocks with the former being nearer the eTh vertex. The points in the northern extremity of the profile are nearer the eTh vertex than the other points on the profile. These visual suggestions were supported by hierarchical cluster analysis, which was able to differentiate between granite and metaclastic outcrops and separate portions of the profile located on different terrains. Portable gamma-ray spectrometry showed, hence, the potential to distinguish granite and metaclastic terrains at a scale useful for engineering works. These results can also be useful for a first comparative zoning of radiological hazards (which are higher for granite).

Application of benzo[a]phenoxazinium chlorides in Antimicrobial Photodynamic Therapy of Candida albicans biofilms.

The use of Antimicrobial Photodynamic Therapy (APDT) as a new approach to treat localized Candida infections is an emerging and promising field nowadays. The aim of this study was to verify the efficacy of photodynamic therapy using two new benzo[a]phenoxazinium photosensitizers against Candida albicans biofilms: N-(5-(3-hydroxypropylamino)-10-methyl-9H-benzo[a]phenoxazin-9-ylidene)ethanaminium chloride (FSc) and N-(5-(11-hydroxyundecylamino)-10-methyl-9H-benzo[a]phenoxazin-9-ylidene)ethanaminium chloride (FSd). The photodynamic activity of dyes against C. albicans biofilms was evaluated by incubating biofilms with dyes in the range of 100-300 µM for 3 or 18 h followed by illumination at 12 or 36 J cm(-2), using a xenon arc lamp (600 ± 2 nm). A total photoinactivation of C. albicans biofilm cells was achieved using 300 µM of FSc with 18 h of incubation, followed by illumination at 36 J cm(-2). Contrarily, FSd had insignificant effect on biofilms inactivation by APDT. The higher uptake of FSc than FSd dye by biofilms during the dark incubation may explain the greater photodynamic effectiveness achieved with FSc. The results obtained stresses out the FSc-mediated APDT potential use to treat C. albicans infections.

Antifungal activity of phenolic compounds identified in flowers from North Eastern Portugal against Candida species.

AIM: To evaluate the antifungal effect of gallic acid, catechin, luteolin and quercetin, phenolic compounds identified from flowers of North Eastern Portugal, against Candida planktonic and biofilm cells. MATERIALS & METHODS: The MICs were determined in Candida planktonic cells and the effect of phenolic compounds on Candida biofilms was assessed through quantification of colony-forming units. RESULTS: MIC values demonstrated that gallic acid presented the highest effect against all Candida species. Catechin showed a similar effect against Candida albicans American Type Culture Collection (ATCC) 90028 cells. In addition, gallic acid and quercetin had demonstrated only a minimal effect against Candida species biofilms. CONCLUSION: Gallic acid affected the growth of the different planktonic Candida species in all concentrations used; still, catechin showed a similar effect against C. albicans ATCC 90028 and Candida glabrata ATCC 2001 cells. In addition, only gallic acid and quercetin demonstrated a slight effect against all Candida species biofilms.

Use and knowledge of Cactaceae in Northeastern Brazil.

BACKGROUND: This study aimed to record the use, and knowledge that residents from São Francisco community (Paraiba, Brazil) have regarding the Cactaceae. METHODS: Semi-structured interviews were carried out with 118 informants; 50 men and 68 women. The cacti cited in this study were organised into use categories and use values were calculated. Differences in the values applied to species and use categories by men and women were compared via a G test (Williams). RESULTS: The nine species identified were: Cereus jamacaru DC., Melocactus bahiensis (Brtitton & Rose) Luetzelb., Nopalea cochenillifera (L.) Salm-Dyck., Opuntia ficus indica (L.) Mill, Opuntia stricta (Haw.) Haw., Pilosocereus gounellei (F.A.C. Weber) Byles & Rowley, Pilosocereus pachycladus F. Ritter, Tacinga inamoena (K. Schum) N.P. Taylor & Stuppy, Tacinga palmadora (Britton & Rose) N.P. Taylor & Stuppy. In total, 1,129 use citations were recorded, divided into 11 categories. The use value categories with the highest scores were forage (0.42), food (0.30) and construction (building) (0.25). P. pachycladus showed the greatest use value, versatility and number of plant parts used. CONCLUSION: The survey showed that the Cactaceae is extremely important for several uses and categories attributed to different species. Apart from contributing to the ethnobotanical knowledge of the Cactaceae, another important focus of this study was to reinforce the necessity for further studies that record the traditional knowledge about this plant family, which has been lost in younger generations.

Importância dos tratamentos complementares na terapêutica cirúrgica do tumor odontogênico queratocísitico / Importance of complementary treatment for the surgical therapy of the keratocystic odontogenic tumor

Tratamentos complementares como excisão tegumentar; ostectomia periférica e aplicação da solução de Carnoy minimizam a recorrência do tumor odontogênico queratocístico após a ecérese conservadora por enucleação ou curetagem, embora sejam desconsiderados por alguns cirurgiões. O artigo revisa as técncias citadas e relata a combinação delas em dois casos dessa neoplasia odontogência, mostrando resultado efetivo em acompanhamento mínimo de cinco anos.

Uso da anestesia tópica na cirurgia de anomalias oculomotoras / Use of topical anaesthesia in the surgery of oculomotor abnormalities

Dezesseis indivíduos adultos portadores de estrabismo de causas diversas foram operados, usando-se a anestesia tópica. A forma de aplicaçäo foi a instilaçäo de colírio de proximetacaína ou proparacaína a 0,5%. Näo ocorreram intercorrências graves e as cirurgias foram realizadas sem grandes dificulades