pH-temperature dual-sensitive nucleolipid-containing stealth liposomes anchored with PEGylated AuNPs for triggering delivery of doxorubicin.

Liposomes (Lip) are useful nanocarriers for drug delivery and cancer nanomedicine because of their ability to efficiently encapsulate drugs with different physical and chemical properties. The pH gradient between normal and tumoral tissues, and their rapid metabolism that induces hyperthermia encourage the development of pH- and thermo-sensitive Lip for delivering anticancer drugs. Nucleolipids have been studied as scaffolding material to prepare Lip, mainly for cancer therapy. Herein, we report for the first time the use of 1,2-dipalmitoyl-sn-glycero-3-(cytidine diphosphate) (DG-CDP) to develop pH/thermo-sensitive nucleolipid-containing stealth Lip stabilized by combination with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol, anchored with NH2-PEGylated gold nanoparticles (PEG-AuNPs, 15 nm) for triggering delivery of doxorubicin (Dox). The optimal composition of DPPC, DG-CDP and cholesterol (94:3:3) was established by Langmuir isotherms. Unloaded and Dox-loaded Lip and AuNPs-Lip exhibited nano-scale sizes (415-650 nm), acceptable polydispersity indexes (<0.33), spherical shapes, and negative Z-potential (-23 to -6.6 mV) due to the phosphate groups of DG-CDP, which allowed the anchoring with positively charged AuNPs. High EE% were achieved (>78%) and although efficient control in the Dox release towards different receptor media was observed, the release of Dox from PEG-AuNPs-Lip-Dox was significantly triggered at acidic pH and hyperthermia conditions, demonstrating its responsiveness to both stimuli. Dox-loaded Lip showed high cytotoxic activity against MDA-MB-231 breast cancer cells and SK-OV-3 ovarian cancer cells, suggesting that Dox was released from these nanocarriers over time. Overall, the liposomal formulations showed promising properties as stimuli-responsive nanocarriers for cancer nanomedicine, with prospects for hyperthermia therapy.

Analgesic and antiinflammatory effects of Nigella orientalis L. seeds fixed oil: Pharmacological potentials and molecular mechanisms.

Nigella species have been widely used in traditional medicine. The aim of this study was to evaluate the antiinflammatory and analgesic potentials of Nigella orientalis L. seeds fixed oil (NOO). The acetic acid writhing test and the formaldehyde-induced licking paw were performed to assess the analgesic activity of the oil. The antiinflammatory activity was first evaluated in vitro by the erythrocyte membrane stabilization then in vivo by xylene- and carrageenan-induced ear and paw edema, respectively. To further understand the molecular mechanism of action of the Nigella extract, lipopolysaccharide-activated RAW 264.7 macrophages were used. Nitric oxide (NO) production was measured by Griess reaction and cell viability by MTT assay. The gene and protein expression of inflammatory mediators were assessed by RT-PCR and western blot, respectively. NOO exerted a potent analgesic effect in in vivo models of writhing test and induced edema. The analyzed molecular mechanisms revealed a role for NO and prostaglandins as molecules mediating the pharmacological effects of the extract through a mechanism involving nuclear factor-κB and mitogen-activated protein kinases. This study demonstrates, for the first time, that the fixed oil of N. orientalis has strong antinociceptive and antiinflammatory properties and might be a promising agent for the treatment of certain inflammation-related diseases.

Pharmacological Extracts and Molecules from Virola Species: Traditional Uses, Phytochemistry, and Biological Activity.

Virola is the largest genus of Myristicaceae in America, comprising about 60 species of medium-sized trees geographically spread from Mexico to southern Brazil. The plant species of this genus have been widely used in folk medicine for the treatment of several ailments, such as rheumatic pain, bronchial asthma, tumors in the joints, intestinal worms, halitosis, ulcers, and multiple infections, due to their pharmacological activity. This review presents an updated and comprehensive summary of Virola species, particularly their ethnomedicinal uses, phytochemistry, and biological activity, to support the safe medicinal use of plant extracts and provide guidance for future research. The Virola spp.'s ethnopharmacology, including in the treatment of stomach pain and gastric ulcers, as well as antimicrobial and tryponosomicidal activities, is attributable to the presence of a myriad of phytoconstituents, such as flavonoids, tannins, phenolic acids, lignans, arylalkanones, and sitosterol. Hence, such species yield potential leads or molecular scaffolds for the development of new pharmaceutical formulations, encouraging the elucidation of not-yet-understood action mechanisms and ascertaining their safety for humans.

Evaluation of Virola oleifera activity in musculoskeletal pathologies: Inhibition of human multiple myeloma cells proliferation and combination therapy with dexamethasone or bortezomib.

ETHNOPHARMACOLOGICAL RELEVANCE: Virola oleifera (Schott) A.C. Smith, Myristicaceae, has been widely used in traditional medicine in Brazil to treat rheumatic pain, joint tumours, skin diseases, halitosis, bronchial asthma, haemorrhoids, and intestinal worms. Recently, research data showed the antioxidant properties in several oxidative stress-related models. However, there is no experimental evidence supporting its potential use in managing rheumatic diseases and bone malignancies. AIMS OF THE STUDY: To evaluate the therapeutic potential of the resin from Virola oleifera in joint and bone diseases, namely arthritis, osteosarcoma, chondrosarcoma, and multiple myeloma. MATERIALS AND METHODS: To determine Virola oleifera resin (VO) effects on arthritis-associated inflammation and cartilage degradation, the LPS-induced NO production, and mRNA and protein expression of ADAMTS5, MMP13, COL2, and ACAN, were evaluated in chondrocytes (ATDC5 and TC28 cell lines). The cytotoxic effects of VO (0.05-50 µg/ml) on multiple myeloma (ARH-77), osteosarcoma (SAOS-2), and chondrosarcoma (SW-1353) cell lines were analysed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The VO effects, combined with dexamethasone or bortezomib, were evaluated in a multiple myeloma cell line. The mechanisms of VO, alone or in combination with bortezomib, were determined by cell cycle analysis through flow cytometry, while expression levels of p-Akt/Akt, p-ERK/ERK, p-p38/p38 MAPK, Bax, Bcl-2, and cleaved-caspase-3/caspase-3 proteins by Western blot. RESULTS: VO had no significant effect on LPS-induced NO production in chondrocytes at non-cytotoxic concentrations. VO treatment diminished the mRNA levels of metalloproteinases and ECM components; however, any significant effect was observed on the protein expression levels. The cell viability of a multiple myeloma cell line was strongly reduced by VO treatment in a dose- and time-dependent manner, while osteosarcoma and chondrosarcoma cell lines viability was significantly affected only by the highest dose assessed. In multiple myeloma cells, VO leads to G2/M cell cycle arrest. Furthermore, it synergizes with dexamethasone by increasing cell toxicity. Finally, VO reverts bortezomib activity by counteracting ERK1/2, Bax, and caspase-3 activation. CONCLUSIONS: The current work supports the ethnopharmacological use of Virola oleifera (Schott) A.C. Smith in bone and joint diseases, but there is no evidence for the amelioration of arthritis-associated inflammatory or catabolic processes. Our data also supports the potential use of Virola oleifera as adjuvant therapy to optimize the pharmacologic effects of current chemotherapeutic drugs. However, possible herb-drug interactions should be considered before clinical application.

Fucoxanthin-Containing Cream Prevents Epidermal Hyperplasia and UVB-Induced Skin Erythema in Mice.

Microalgae represent a source of bio-active compounds such as carotenoids with potent anti-inflammatory and antioxidant properties. We aimed to investigate the effects of fucoxanthin (FX) in both in vitro and in vivo skin models. Firstly, its anti-inflammatory activity was evaluated in LPS-stimulated THP-1 macrophages and TNF-α-stimulated HaCaT keratinocytes, and its antioxidant activity in UVB-irradiated HaCaT cells. Next, in vitro and ex vivo permeation studies were developed to determine the most suitable formulation for in vivo FX topical application. Then, we evaluated the effects of a FX-containing cream on TPA-induced epidermal hyperplasia in mice, as well as on UVB-induced acute erythema in hairless mice. Our results confirmed the in vitro reduction of TNF-α, IL-6, ROS and LDH production. Since the permeation results showed that cream was the most favourable vehicle, FX-cream was elaborated. This formulation effectively ameliorated TPA-induced hyperplasia, by reducing skin edema, epidermal thickness, MPO activity and COX-2 expression. Moreover, FX-cream reduced UVB-induced erythema through down-regulation of COX-2 and iNOS as well as up-regulation of HO-1 protein via Nrf-2 pathway. In conclusion, FX, administered in a topical formulation, could be a novel natural adjuvant for preventing exacerbations associated with skin inflammatory pathologies as well as protecting skin against UV radiation.

Association of metreleptin treatment and dietary intervention with neurological outcomes in Celia's encephalopathy.

Celia's encephalopathy (progressive encephalopathy with/without lipodystrophy, PELD) is a recessive neurodegenerative disease that is fatal in childhood. It is caused by a c.985C>T variant in the BSCL2/seipin gene that results in an aberrant seipin protein. We evaluated neurological development before and during treatment with human recombinant leptin (metreleptin) plus a dietary intervention rich in polyunsaturated fatty acids (PUFA) in the only living patient. A 7 years and 10 months old girl affected by PELD was treated at age 3 years with metreleptin, adding at age 6 omega-3 fatty acid supplementation. Her mental age was evaluated using the Battelle Developmental Inventory Screening Test (BDI), and brain PET/MRI was performed before treatment and at age 5, 6.5, and 7.5 years. At age 7.5 years, the girl remains alive and leads a normal life for her mental age of 30 months, which increased by 4 months over the last 18 months according to BDI. PET images showed improved glucose uptake in the thalami, cerebellum, and brainstem. This patient showed a clear slowdown in neurological regression during leptin replacement plus a high PUFA diet. The aberrant BSCL2 transcript was overexpressed in SH-SY5Y cells and was treated with docosahexaenoic acid (200 µM) plus leptin (0.001 mg/ml) for 24 h. The relative expression of aberrant BSCL2 transcript was measured by qPCR. In vitro studies showed significant reduction (32%) in aberrant transcript expression. This therapeutic approach should be further studied in this devastating disease.

Analytical determination of flavonoids aimed to analysis of natural samples and active packaging applications.

Several HPLC and UHPLC developed methods were compared to analyse the natural antioxidants catechins and quercetin used in active packaging and functional foods. Photodiode array detector coupled with a fluorescence detector and compared with LTQ-Orbitrap-MS was used. UHPLC was investigated as quick alternative without compromising the separation, analysis time shortened up to 6-fold. The feasibility of the four developed methods was compared. Linearity up to 0.9995, low detection limits (between 0.02 and 0.7 for HPLC-PDA, 2 to 7-fold lower for HPLC- LTQ-Orbitrap-MS and from 0.2 to 2mgL(-)(1) for UHPLC-PDA) and good precision parameters (RSD lower than 0.06%) were obtained. All methods were successfully applied to natural samples. LTQ-Orbitrap-MS allowed to identify other analytes of interest too. Good feasibility of the methods was also concluded from the analysis of catechin and quercetin release from new active packaging materials based on polypropylene added with catechins and green tea.

Enhancing the release of the antioxidant tocopherol from polypropylene films by incorporating the natural plasticizers lecithin, olive oil, or sunflower oil.

In this work, natural plasticizers-modified polypropylenes intended for food active packaging were developed. Sunflower oil, olive oil, and soy lecithin, without any known harmful effects or toxicity, were employed as natural plasticizers, also implementing the attractiveness of using synthetic plastics on active packaging developments. Their incorporation during the extrusion of polypropylene was tried as a means to obtain polymers with improved diffusion paths, allowing differences in antioxidant release rates for active packaging materials. Thermal and rheological characterization of the films showed that blending natural plasticizers do not significantly modify their thermal properties; however, small variations of viscoelastic properties were observed. Furthermore, the resulting release of tocopherol was highly dependent on the polymer formulation. Furthermore, it was clearly time-controlled by using those natural plasticizers, especially olive oil. Antioxidant activity results also showed that packaged foods are protected against oxidative degradation over time, resulting from the improved release of the antioxidants.

Improving the capacity of polypropylene to be used in antioxidant active films: incorporation of plasticizer and natural antioxidants.

Two types of active antioxidant food packages with improved release properties, based on polypropylene (PP) as one of the most common polymers used in food-packaging applications, were developed. Incorporation of catechin and green tea as antioxidant provided PP with 6 times higher stabilization against thermal oxidation. Release of natural antioxidants (catechins, gallic acid, caffeine, and quercetin) into various food simulants from that nonpolar matrix were improved by blending poly(propylene glycol)-block-poly(ethylene glycol)-block-poly(propylene glycol) (PPG-PEG-PPG) as plasticizer into the polymer formulation. Increasing release levels between 10- and 40-fold into simulant A and between 6 and 20-fold into simulant D1 resulted from the incorporation of catechin and green tea as antioxidants and PPG-PEG-PPG as plasticizer into the film formulation. The efficiency of the antioxidants in the food simulants after the release process was also corroborated through antioxidant activity tests. Therefore, the developed PPG-PEG-PPG-modified polypropylene resulted in a potential system to be used in active packaging.

Effectiveness of the EndoActivator System in removing the smear layer after root canal instrumentation.

INTRODUCTION: Elimination of the smear layer after root canal instrumentation requires the use of irrigating solutions. This cleaning can be completed with passive ultrasonic or sonic irrigation. The aim of this study was to evaluate the effectiveness of the EndoActivator System in removing the smear layer after rotary root canal instrumentation, with and without a final flush of 17% ethylenediaminetetraacetic acid (EDTA) solution, in coronal, middle, and apical thirds. METHODS: Forty single-canal teeth were decoronated and randomly divided into 4 groups (n = 10). The groups were instrumented by using Mtwo System. EndoActivator was used with a final rinse of 1 mL of 17% EDTA or 4% NaOCl for 1 minute. The roots were longitudinally split and were grooved in the coronal, middle, and apical thirds. Scanning electron microscopy digital photomicrographs at 400x were taken to evaluate the amount of smear layer in each third. RESULTS: The NaOCl/EndoActivator group did not remove any smear layer of the root canal wall (100% in the coronal, middle, and apical thirds). In the groups that used 17% EDTA (with or without EndoActivator), the smear layer was eliminated completely (100%) in the coronal third, but the amount of removal was less in the other two thirds. The comparisons between NaOCl versus NaOCl/EndoActivator groups and EDTA/NaOCl versus EDTA/EndoActivator/NaOCl groups showed no significant differences in root canal thirds. CONCLUSIONS: The EndoActivator System did not enhance the removal of smear layer as compared with conventional Max-I-Probe irrigation with NaOCl and EDTA.

Efectividad del tratamiento homeopático con la sheisi en la crisis hipertensiva / Effectiveness of Homeopathic Treatment with Lasheisi in Hypertensive Crisis

Se realizó un estudio descriptivo, prospectivo, a 95 pacientes portadores de hipertensión arterial descompensada tomados de forma aleatoria, a nivel del cuerpo de guardia del policlínico Jimmy Hirtzel de la ciudad de Bayamo en un periodo de un año con el objetivo de evaluar la efectividad del tratamiento homeopático en la hipertensión arterial descompensada, nuestro universo de trabajo estuvo constituido por todos los pacientes hipertensos descompensados que llegaron al cuerpo de guardia en ese periodo de tiempo y nuestra muestra todos aquellos que aceptaron incluirse en la investigación (95 pacientes), posteriormente se le aplico el tratamiento homeopático con lashesi a la 30 Ch a todos los pacientes que entraron en el estudio a razón de 10 gotas s/l cada 15 minutos hasta tres dosis, las principales variables que se analizaron fueron: Edad, sexo, tiempo de compensación, dosis utilizadas, grado de satisfacción y efectividad del tratamiento; los 95 pacientes estuvieron de acuerdo a participar en el estudio y reunían las condiciones de los criterios de inclusión, la mejor respuesta terapéutica estuvo vinculada con aquellos pacientes pertenecientes al grupo de edades ente 40 y 69 años, la primera dosis aplicada reunió el mayor por ciento de pacientes compensados, con un predominio en el sexo femenino, de 95 paciente que se le aplico el tratamiento homeopático 84 de ellos se controlaron con esta terapéutica y solo 11 no lo lograron, el grado de satisfacción fue de muy bueno ya que el 88.4 por ciento de los pacientes la aceptaron como buena, inocua y poco costosa(AU)

It was performed a descriptive prospective study to 95 patients carriers of decompensated high blood pressure taken at random, at the guard room of Jimmy Hirzel Polyclinic in a period of a year with the objective of evaluating the effectiveness of homeopathy treatment in uncontrolled high blood pressure, the universe was constituted by all uncontrolled hypertensive patients that came into the facility, and the sample was composed by all the patients that agreed to participate in the research. Afterwards there was applied homeopathy treatment with lashisi to 30 ch to all patients that took part with 10 drops s/l each 15 minutes until three doses. The variables used were age, sex, time of compensation, used dose, level of satisfaction and treatment effectiveness. The best therapeutic response was associated with those patients belonging to the group ages among 40 to 69 years old, the first dose applied gathered the greater percentage of controlled patients, prevailing female. Eighty four patients were controlled with this medical and only 11 patients could not get it. The level of satisfaction was good, 88.4 per cent accepted as innocuous and less expensive(AU)

The effectiveness of different acid irrigating solutions in root canal cleaning after hand and rotary instrumentation.

The goal of this study was to evaluate the cleaning ability of three acid irrigating solutions after hand and rotary instrumentation. Eighty human teeth were randomly divided in eight groups. Four groups were prepared with hand instrumentation and other four with ProTaper. The irrigating solutions were 15% citric acid plus 2.5% NaOCl; 15% EDTA plus 2.5% NaOCl; 5% orthophosphoric acid plus 2.5% NaOCl; and 2.5% NaOCl alone as control. Canal walls were observed with scanning electron microscopy, and photomicrographs were taken in apical, middle, and coronal thirds. A scoring system for debris and smear layer was used. Acid solutions with 2.5% NaOCl were effective in the elimination of smear layer or debris, and no significant differences were showed in smear layer removal between techniques. However, 2.5% NaOCl did not remove smear layer or debris, and no significant differences in debris were observed between manual and rotary techniques.