Characterization of the cicatrization process in diabetic foot ulcers based on the production of reactive oxygen species

The present study aims at evaluating the correlation between the free radical formation and the healing action of lower limbs’ ulcers in a randomized controlled trial with the use of an adhesive derived from natural latex associated with a light-emitting diode (LED) circuit. The sample consists of 15 participants with lower limb lesions divided into three groups: group 1 case (5 participants) received the proposed dressing system adhesive of the natural latex associated with the LED circuit; group 2 control (5 participants) received the dressings at home performed by nurses according to and established by the clinic of wounds (treated with calcium alginate or silver foam); and group 3 (5 participants) also received the dressing in their homes with the use of the dressing adhesive derived from the natural latex associated with the LED circuit. The collected data were analyzed qualitatively and quantitatively by electron paramagnetic resonance for determination of free radical formation. Kruskal-Wallis statistical test was used to evaluate the effect of treatment on the lower limb’s ulcer cicatrization process and its correlation with free radical. The results obtained corroborated the hypothesis about the reduction of the quantity of these molecules in the end of treatment related to the healing wound

Antioxidant Activity and Stable Free Radicals in Robusta Green Coffee Genotypes/ Atividade antioxidante e Radicais Estáveis Livres em Genótipos de Café Verde Robusta

This study reports on physicochemical properties (total soluble solids, titratable acidity plus caffeine, trigonelline, 5-CQA, and total phenols contents) of five genotypes of Robusta coffee beans; Bamburral, Beira Rio, Clementino, Coringa, and Pirata. Green bean of Clementino presents the highest concentration of total soluble solids in dry basis (18.0%) and the lowest titratable acidity (154.0 mL-NaOH). Moreover, green beans of Clementino and Pirata show intermediate values of caffeine and trigonelline contents while presenting the highest yield index ofstable free radical formation after roasting, respectively equal to 26 and 23 (electron paramagnetic resonance-EPR data), and the highest content of 5-CQA (around 42 mg/g). Green beans of Bamburral, Beira Rio and Coringa show the highest concentration of total phenols (53 - 56 mg/g), meaning the highest antioxidant activities (IC50 test) but reduced yield index of formation of stable free radicals after roasting, respectively equal to 13.0, 5.9, and 13.0 (EPR data)

Origem dos momentos magnéticos localizados em ligas de cromo com metais de transição por oscilações de Friedel

The fundamental magnetic properties of chromium and its alloys Cr(1¡x)Nb(x) and Cr(1¡x)V(x), were investigated with the aim to achieve a better comprehension about the nature of the spin density waves. Was dened the edge between the AF1 and AF2 phases on the Cr(1¡x)Nb(x) alloys as function of niobium concentration. Using x-rays di®raction techniques was studied de formation of antiferromagnetic domains in the surfaces with [001] direction of chromium samples. As observed, cooling the sample under applied magneticeld do not create new magnetic domains, actuating only on its reorien- tation. The magnetic history of sample does not in°uence the surface magnetic domains formation, been an exclusive property of the sample surface crystalographic orientation. Was observed the formation of localized magnetic moment on paramagnetic phase of Cr(1¡x)Nb(x) alloys. This phenomenon early studied in Cr(1¡x)V(x) alloys, shown important qualitative di®erences which are explained on Friedel oscillations picture generated by impurity atoms. Was realized x-rays di®raction experiments which provided evidences of the existence of Friedel oscillations in Cr(1¡x)V(x) alloys.Financiadora de Estudos e ProjetosAs propriedades magnéticas fundamentais do cromo e de suas ligas de Cr(1¡x)Nb(x) e Cr(1¡x)V(x), foram investigadas com o objetivo de melhorar a compreensão sobre a natureza das ondas de densidade de spin (spin-density-wave). Foi determinada a fronteira entre as fases AF1 e AF2 nas ligas Cr(1¡x)Nb(x) em função da concentração de nióbio. Utilizando tecnica de difração de raios-x foi estudada a formação dos dominios antiferro-magnéticos nas superfícies paralelas às direções [001] e [111] de uma amostra de cromo crescida por transporte qumico de vapor (chemical vapor deposition method). Os resultados obtidos com o estado de monodomínio volumétrico preparado por resfriamento das amostras com campo magnético aplicado, comparados com aqueles das amostras em estado multi-domínio volumétrico e monodomínio de superfície, levam a conclusão de que a aplicação de campo magnético ou de outra maneira, a história magnética da amostra, não implica na formação de novos domínios magnéticos na superfície do cromo e que o campo magnético atua especificamente na reorientação do domínios existentes. Portanto, a formação de monodomínios superficiais e uma propriedade exclusiva da orientação cristalografica da superfície da amostra. Foi observada a formação de momento magnético localizado na fase paramagnética das ligas de Cr(1¡x)Nb(x). Esse fenômeno, anteriormente estudado nas ligas Cr(1¡x)V(x), apresentou diferenças qualitativas importantes na sua manifestação nas ligas de Cr(1¡x)Nb(x), que são explicadas a partir das oscilações de Friedel que se formam na região dos átomo de vanadio e nióbio. Foram realizados experimentos de difração de raios-x que forneceram evidências da existência das oscilações de Friedel nas ligas Cr(1¡x)V(x)

AlPc/ZnPc-based oncological photodynamic therapy for a selective eradication of leukemic cells from ovarian tissue.

Due to the risk of reintroducing malignant cells, autotransplantation of cryopreserved ovarian tissue is not allowed in leukemia patients. In order to restore fertility in these patients, ex vivo purging of ovarian fragments could be proposed as a strategy to eradicate malignant cells before grafting. Photodynamic therapy (PDT), as a clinical-approved modality, is a minimally invasive and selective therapeutic for eliminating malignant cells. The present work aims therefore to evaluate the phototoxicity of two photosensitizers (aluminum phthalocyanine (AlPc) and zinc phthalocyanine (ZnPc)) on leukemia cells. To this end, two lines of malignant cells (K562 and HL-60) and isolated ovarian stromal cells (control) were treated by PDT using a diode laser with various energy densities. Cell viability after the treatment, the amount of generated reactive oxygen species, dark toxicity of the photosensitizers, and single-cell morphology were studied. Our results demonstrated that using irradiation with the energy density of 10 J/cm, 1 µM AlPc could significantly reduce the viability of K562 (4.73 ± 0.14%) and HL-60 (2.74 ± 0.31%). Similarly, the viability of these cells was reduced (K562 cells: 3.84 ± 0.81%; HL-60 cells: 6.82 ± 3.21%) with 1 µM ZnPc and an energy density of 50 J/cm. On the other hand, these PDT protocols had no significant effect on stromal cells. These findings indicate that our approach can be a promising strategy for the safe restoration of fertility in leukemia patients. However, further studies are necessary to assess its efficiency in ovarian fragments containing malignant cells to determine their eradication rate and the effect of our treatment on the survival of stromal cells and preantral follicles

Cytoglobin attenuates neuroinflammation in lipopolysaccharide-activated primary preoptic area cells via NF-κB pathway inhibition

Cytoglobin (Cygb) is a hexacoordinate protein, associated with the transport of oxygen, nitric oxide scavenging, tumor suppression and protection against oxidative stress and inflammation. This protein is expressed in brain areas including the preoptic area (POA) of the anterior hypothalamus, the region responsible for the regulation of body temperature. In this study, we show that Cygb is upregulated in the rat hypothalamus 2.5 h and 5 h after intravenous administration of lipopolysaccharide (LPS). We investigated the effect of treatment with Cygb in POA primary cultures stimulated with LPS for 4 h. The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were measured and the results showed that Cygb reduced the concentrations of both cytokines. We further observed a decrease in immunoreactivity of the inflammatory transcription factor nuclear factor-κB (NF-κB), but not NF-IL6 and STAT3, in the nucleus of Cygb-treated POA cells. These findings suggest that Cygb attenuates the secretion of IL-6 and TNF-α in LPS-stimulated POA primary cultures via inhibition of the NF-κB signaling pathway, indicating that this protein might play an important role in the control of neuroinflammation and fever

Photodynamic therapy directed to melanoma skin cancer by thermosensitive hydrogel containing Chlorophyll A

Melanoma, a severe form of skin cancer intricately linked to genetic and environmental factors, is predicted to reach 100,000 new cases worldwide by 2040, underscoring the need for effective and safe treatment options. In this study, we assessed the efficacy of a photosensitizer called Chlorophyll A (Chl-A) incorporated into hydrogels (HGs) made of chitosan (CS) and poloxamer 407 (P407) for Photodynamic Therapy (PDT) against the murine melanoma cell line B16-F10. The HG was evaluated through various tests, including rheological studies, SEM, and ATR-FTIR, along with cell viability assays. The CS- and P407-based HGs effectively released Chl-A and possessed the necessary properties for topical application. The photodynamic activity of the HG containing Chl-A was evaluated in vitro, demonstrating high therapeutic potential, with an IC50 of 25.99 µM—an appealing result when compared to studies in the literature reporting an IC50 of 173.8 µM for cisplatin, used as a positive control drug. The developed formulation of CS and P407-based HG, serving as a thermosensitive system for topical applications, successfully controlled the release of Chl-A. In vitro cell studies associated with PDT exhibited potential against the melanoma cell line.Instituto de Ciências Biológicas (IB)Departamento de Genética e Morfologia (IB GEM)Departamento de Biologia Celular (IB CEL)Instituto de Física (IF)Faculdade UnB Ceilândia (FCE)Colegiado de Bases Biológicas e da Saúde (FCE-BASES

Mitochondrial pyruvate carrier as a key regulator of fever and neuroinflammation

The mitochondrial pyruvate carrier (MPC) is an inner-membrane transporter that facilitates pyruvate uptake from the cytoplasm into mitochondria. We previously reported that MPC1 protein levels increase in the hypothalamus of animals during fever induced by lipopolysaccharide (LPS), but how this increase contributes to the LPS responses remains to be studied. Therefore, we investigated the effect of UK 5099, a classical MPC inhibitor, in a rat model of fever, on hypothalamic mitochondrial function and neuroinflammation in LPS-stimulated preoptic area (POA) primary microcultures. Intracerebroventricular administration of UK 5099 reduced the LPS-induced fever. High-resolution respirometry revealed an increase in oxygen consumption and oxygen flux related to ATP synthesis in the hypothalamic homogenate from LPS-treated animals linked to mitochondrial complex I plus II. Preincubation with UK 5099 prevented the LPS-induced increase in oxygen consumption, ATP synthesis and spare capacity only in complex I-linked respiration and reduced mitochondrial H2O2 production. In addition, treatment of rat POA microcultures with UK 5099 reduced the secretion of the proinflammatory and pyrogenic cytokines TNFα and IL-6 as well as the immunoreactivity of inflammatory transcription factors NF-κB and NF-IL6 four hours after LPS stimulation. These results suggest that the regulation of mitochondrial pyruvate metabolism through MPC inhibition may be effective in reducing neuroinflammation and fever

Nanographene oxide-methylene blue as phototherapies platform for breast tumor ablation and metastasis prevention in a syngeneic orthotopic murine model

Abstract Background In the photodynamic therapy (PDT), the photosensitizer absorbs light and transfers the energy of the excited state to the oxygen in the cell environment producing reactive oxygen species (ROS), that in its turn, may cause cell damage. In the photothermal therapy (PTT), light also is responsible for activating the photothermal agent, which converts the absorbed energy in heat. Graphene oxide is a carbon-based material that presents photothermal activity. Its physical properties allow the association with the photosensitizer methylene blue and consequently the production of ROS when submitted to light irradiation. Therefore, the association between nanographene oxide and methylene blue could represent a strategy to enhance therapeutic actions. In this work, we report the nanographene oxide-methylene blue platform (NanoGO-MB) used to promote tumor ablation in combination with photodynamic and photothermal therapies against a syngeneic orthotopic murine breast cancer model. Results In vitro, NanoGO-MB presented 50% of the reactive oxygen species production compared to the free MB after LED light irradiation, and a temperature increase of ~ 40 °C followed by laser irradiation. On cells, the ROS production by the nanoplatform displayed higher values in tumor than normal cells. In vivo assays demonstrated a synergistic effect obtained by the combined PDT/PTT therapies using NanoGO-MB, which promoted complete tumor ablation in 5/5 animals. Up to 30 days after the last treatment, there was no tumor regrowth compared with only PDT or PTT groups, which displayed tumoral bioluminescence 63-fold higher than the combined treatment group. Histological studies confirmed that the combined therapies were able to prevent tumor regrowth and liver, lung and spleen metastasis. In addition, low systemic toxicity was observed in pathologic examinations of liver, spleen, lungs, and kidneys. Conclusions The treatment with combined PDT/PTT therapies using NanoGO-MB induced more toxicity on breast carcinoma cells than on normal cells. In vivo, the combined therapies promoted complete tumor ablation and metastasis prevention while only PDT or PTT were unable to stop tumor development. The results show the potential of NanoGO-MB in combination with the phototherapies in the treatment of the breast cancer and metastasis prevention

Novel iron oxide nanocarriers loading finasteride or dutasteride : enhanced skin penetration for topical treatment of alopecia

In the present study, iron oxide nanoparticles, in the form of maghemite core coated with lauric acid (ION), were synthesized and loaded with finasteride (FIN) or dutasteride (DUT) as a novel drug delivery system for the topical treatment of alopecia. Additionally, developed formulations (FIN-ION and DUT-ION) were completely elaborated with components involved in the follicle metabolism, i.e., lauric acid, which acts as a 5α-reductase inhibitor, and iron which deficiency has been related to hair loss aggravation. Stability assessment conducted over the course of 90 days showed they are highly stable, with pH 7.4, constant EE% (>99%), and practically unchanged particle size and zeta potential. Besides drug distribution, the actual number of iron oxide nanoparticles, through a newly developed method using ferromagnetic resonance, was determined in each skin layer following permeation experiments. Despite the same donor concentration of colloids, nanoparticle distribution in the skin varied according to the loaded molecule. While DUT did not interfere with the nanoparticle natural tendency to accumulate within the hair follicle shafts, FIN presence hampered nanosystem interaction with the skin. Still, both formulations provided a higher skin drug penetration, compared to each respective control solution. Additionally, iron nanocarriers present a desirable visual characteristic, as the dark color aspect might instantly help disguise scarce hair follicle areas. These findings suggest the nanoformulations are highly promising for alopecia therapies

Development of New Natural Lipid-Based Nanoparticles Loaded with Aluminum-Phthalocyanine for Photodynamic Therapy against Melanoma

Photodynamic therapy (PDT) mediated by photosensitizers loaded in nanostructures as solid lipid nanoparticles has been pinpointed as an effective and safe treatment against different skin cancers. Amazon butters have an interesting lipid composition when it comes to forming solid lipid nanoparticles (SLN). In the present report, a new third-generation photosensitizing system consisting of aluminum-phthalocyanine associated with Amazon butter-based solid lipid nanoparticles (SLN-AlPc) is described. The SLN was developed using murumuru butter, and a monodisperse population of nanodroplets with a hydrodynamic diameter of approximately 40 nm was obtained. The study of the permeation of these AlPc did not permeate the analyzed skin, but when incorporated into the system, SLN-AlPc allowed permeation of almost 100% with 8 h of contact. It must be emphasized that SLN-AlPc was efficient for carrying aluminum-phthalocyanine photosensitizers and exhibited no toxicity in the dark. Photoactivated SLN-AlPc exhibited a 50% cytotoxicity concentration (IC50) of 19.62 nM when applied to B16-F10 monolayers, and the type of death caused by the treatment was apoptosis. The exposed phospholipid phosphatidylserine was identified, and the treatment triggered a high expression of Caspase 3. A stable Amazon butter-based SLN-AlPc formulation was developed, which exhibits strong in vitro photodynamic activity on melanoma cells