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1.
PLoS One ; 13(9): e0203748, 2018.
Article in English | MEDLINE | ID: mdl-30212521

ABSTRACT

Violacein is a violet pigment produced by Chromobacterium violaceum that possesses several functions such as antibacterial, antiviral, antifungal, and antioxidant activities. The search for potential compounds and therapies that may interfere with and modulate the gut microbial consortia without causing severe damage and increased resistance is important for the treatment of inflammatory, allergic, and metabolic diseases. The aim of the present work was to evaluate the ability of violacein to change microbial patterns in the mammalian gut by favoring certain groups over the others in order to be used as a therapy for diseases associated with changes in the intestinal microflora. To do this, we used male Wistar rats, and administered violacein orally, in low (50 µg/ml) and high (500 µg/ml) doses for a month. Initially, the changes in the microbial diversity were observed by DGGE analyses that showed that the violacein significantly affects the gut microbiota of the rats. Pyrosequencing of 16S rDNA was then employed using a 454 GS Titanium platform, and the results demonstrated that higher taxonomic richness was observed with the low violacein treatment group, followed by the control group and high violacein treatment group. Modulation of the microbiota at the class level was observed in the low violacein dose, where Bacilli and Clostridia (Firmicutes) were found as dominant. For the high violacein dose, Bacilli followed by Clostridia and Actinobacteria were present as the major components. Further analyses are crucial for a better understanding of how violacein affects the gut microbiome and whether this change would be beneficial to the host, providing a framework for the development of alternative treatment strategies for intestinal diseases using this compound.


Subject(s)
Anti-Bacterial Agents/pharmacology , Chromobacterium/chemistry , Gastrointestinal Microbiome/drug effects , Indoles/pharmacology , Administration, Oral , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Bacillus/genetics , Bacillus/isolation & purification , Bacteria/genetics , Bacteria/isolation & purification , Chromobacterium/metabolism , High-Throughput Nucleotide Sequencing , Indoles/chemistry , Indoles/isolation & purification , Intestines/microbiology , Male , RNA, Ribosomal, 16S/chemistry , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/metabolism , Rats , Rats, Wistar , Sequence Analysis, DNA
2.
Burns ; 44(2): 359-369, 2018 03.
Article in English | MEDLINE | ID: mdl-29032978

ABSTRACT

BACKGROUND: Albino hairless mouse (AHM) has been used as a biological model in photodermatology. However, the experimental landscape is diverse to follow and need particular attention. PURPOSE: Irradiation parameters were investigated for the development of a protocol to assess alterations in the AHM skin using Simulated Solar Light (SSL). The present study was compared with published articles (last 15 years) according to irradiation protocols, morphological findings to minimize animal suffering and UV exposure. MATERIALS AND METHODS: Three groups: Control (G1), experimental - sunburn (G2) and skin photodamage assay (G3). G2 were immobilized and exposed to SSL once for 15, 30 and 45min. G3 were exposed to SSL, without immobilization, for 15min once a day for one week. The dorsal skin was analyzed using hematoxylin and eosin technique. RESULTS: G2 displayed different sunburn degrees. Based on the profile of the observed morphological alterations, a 15min irradiation was chosen as the exposure time to expose G3, without immobilization, for 5 consecutive days. CONCLUSION: These conditions produced the same morphological changes in the AHM with a shorter solar exposure time, without immobilizing the animals but using environmental exposure fluences, conforming to 3R (reduction - refinement - replacement) recommendations.


Subject(s)
Skin/radiation effects , Sunburn/pathology , Sunlight/adverse effects , Ultraviolet Rays/adverse effects , Animals , Male , Mice , Mice, Hairless , Restraint, Physical , Skin/pathology
3.
Life Sci ; 158: 7-13, 2016 Aug 01.
Article in English | MEDLINE | ID: mdl-27328417

ABSTRACT

AIMS: Cancer cells produce higher amounts of reactive oxygen species (ROS) than their normal counterparts. It has been suggested that a further increase in ROS concentration in these cells would lead to oxidative damage-driven death. Thus, we aimed to understand how the intra- and extracellular redox homeostasis differences set cell death response to ROS in breast cancer cell lines. MAIN METHODS: Intra- and extracellular ROS generation was evaluated in tumoral (MCF-7 and MDA-MB-231) and non-tumoral (MCF10A) breast epithelial cells, as well as H2O2 concentration in the culture medium, glutathione peroxidase (GPx), total superoxide dismutase (SOD) and catalase activities, extracellular H2O2 scavenging capacity and total thiol content. Cell viability was determined after H2O2 exposure using the MTT assay. KEY FINDINGS: We have found an increased extracellular ROS production in tumor cells when compared to the non-tumoral lineage. MCF10A cells had higher H2O2 concentration in the extracellular medium. Moreover, extracellular H2O2-scavenging activity was higher in MDA-MB-231 when compared to MCF10A and MCF-7. Regarding intracellular antioxidant activity, a lower GPx activity in tumor cell lines and a higher catalase activity in MDA-MB-231 were observed. Thiol content was lower in MDA-MB-231. Additionally, tumor cell lines were more sensitive to H2O2 exposure than the non-tumoral cells. SIGNIFICANCE: The present report shows that the capability to generate and metabolize ROS differ greatly among the breast cancer cell lines, thus suggesting that redox balance is finely regulated during carcinogenesis. Therefore, our data suggest that therapeutic approaches targeting the redox status might be useful in the treatment of breast tumors.


Subject(s)
Breast Neoplasms/metabolism , Cell Death/drug effects , Homeostasis , Hydrogen Peroxide/pharmacology , Breast Neoplasms/pathology , Cell Line, Tumor , Female , Glutathione Peroxidase/genetics , Humans , Oxidation-Reduction , RNA, Messenger/metabolism , Reactive Oxygen Species/metabolism , Superoxide Dismutase/genetics
4.
Microb Ecol ; 65(2): 325-35, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23001596

ABSTRACT

Martian surface microbial inhabitants would be challenged by a constant and unimpeded flux of UV radiation, and the study of analog model terrestrial environments may be of help to understand how such life forms could survive under this stressful condition. One of these environments is the Atacama Desert (Chile), a well-known Mars analog due to its extreme dryness and intense solar UV radiation. Here, we report the microbial diversity at five locations across this desert and the isolation of UVC-tolerant microbial strains found in these sites. Denaturing gradient gel electrophoresis (DGGE) of 16S rDNA sequences obtained from these sites showed banding patterns that suggest distinct and complex microbial communities. Analysis of 16S rDNA sequences obtained from UV-tolerant strains isolated from these sites revealed species related to the Bacillus and Pseudomonas genera. Vegetative cells of one of these isolates, Bacillus S3.300-2, showed the highest UV tolerance profile (LD(10) = 318 J m(2)), tenfold higher than a wild-type strain of Escherichia coli. Thus, our results show that the Atacama Desert harbors a noteworthy microbial community that may be considered for future astrobiological-related research in terms of UV tolerance.


Subject(s)
Bacteria/isolation & purification , Bacteria/radiation effects , Desert Climate , Soil Microbiology , Bacteria/classification , Bacteria/genetics , Biodiversity , Chile , DNA, Bacterial/genetics , Denaturing Gradient Gel Electrophoresis , Exobiology , RNA, Ribosomal, 16S/genetics , Radiation Tolerance , Ultraviolet Rays
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