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1.
Neotrop. ichthyol ; 19(1): e200102, 2021. tab, graf, ilus, mapas
Article in English | LILACS, VETINDEX | ID: biblio-1154969

ABSTRACT

A new species of Hyphessobrycon belonging to the Hyphessobrycon heterorhabdus species-group from the lower rio Tapajós, state of Pará, Brazil, is described. The new species is allocated into the Hyphessobrycon heterorhabdus species-group due to its color pattern, composed by an anteriorly well-defined, horizontally elongated humeral blotch that becomes diffuse and blurred posteriorly, where it overlaps with a conspicuous midlateral dark stripe that becomes blurred towards the caudal peduncle and the presence, in living specimens, of a tricolored longitudinal pattern composed by a dorsal red or reddish longitudinal stripe, a middle iridescent, golden or silvery longitudinal stripe, and a more ventrally-lying longitudinal dark pattern composed by the humeral blotch and dark midlateral stripe. It can be distinguished from all other species of the group by possessing humeral blotch with a straight or slightly rounded ventral profile, lacking a ventral expansion present in all other species of the group. The new species is also distinguished from Hyphessobrycon heterorhabdus by a 9.6% genetic distance in the cytochrome c oxidase I gene. The little morphological distinction of the new species when compared with its most similar congener, H. heterorhabdus, indicates that the new species is one of the first truly cryptic fish species described from the Amazon basin.(AU)


Uma nova espécie de Hyphessobrycon pertencente ao grupo Hyphessobrycon heterorhabdus é descrita da região do baixo rio Tapajós, estado do Pará, Brasil. A nova espécie é incluída no grupo Hyphessobrycon heterorhabdus devido ao seu padrão de coloração, composto por uma mancha umeral alongada, anteriormente bem definida, que se torna difusa e borrada posteriormente, onde se sobrepõe a uma conspícua faixa escura médio-lateral que se torna borrada próxima ao pedúnculo caudal, e pela presença, em exemplares vivos, de um padrão longitudinal tricolor, composto por uma faixa longitudinal vermelha ou avermelhada dorsal, uma faixa média iridescente dourada ou prateada e, mais ventralmente, o padrão longitudinal escuro composto pela faixa escura médio-lateral e mancha umeral. A espécie pode ser distinguida das outras espécies pertencentes ao grupo por possuir uma mancha umeral com região ventral retilínea ou levemente arredondada, sem uma expansão ventral presente nas demais espécies do grupo. A espécie também se diferencia de Hyphessobrycon heterorhabdus por uma distância genética de 9,6% no gene citocromo c oxidase I. A sutil diferença morfológica da nova espécie quando comparada ao seu congênere mais similar, H. heterorhabdus, indica que a nova espécie é uma das primeiras espécies de peixes verdadeiramente crípticas descritas da Bacia Amazônica.(AU)


Subject(s)
Animals , DNA , Cytochromes c , Biodiversity , Characidae , Amazonian Ecosystem
2.
Article in Chinese | WPRIM | ID: wpr-826660

ABSTRACT

OBJECTIVE@#To observe the impacts of electroacupuncture (EA) on neurological function, the pathological morphology in brain tissue, apoptosis level and the protein expressions of apoptosis-related cytochrome C (Cyt-C) and cysteine aspartic acid protease-9 (Caspase-9) in the rats with traumatic brain injury (TBI) and explore the potential mechanism of EA in treatment of TBI.@*METHODS@#A total of 70 clean-grade SD mice were randomized into a blank group (8 rats), a sham-operation group (8 rats), a model group (27 rats) and an EA group (27 rats). In terms of interventions of 3, 7 and 14 days, 3 subgroups were divided in the model group and the EA group successively, 9 rats in each subgroup. The modified Feeney free-fall percussion method was adopted to establish TBI models of rats. In the sham-operation group, only the skull was exposed and drilled and no free-fall percussion was exerted. One day after modeling, EA was given in the rats of EA group at "Shuigou" (GV 26), "Baihui" (GV 20) and "Neiguan" (PC 6) and "Zusanli" (ST 36) on the affected side, with intermittent wave, 2 Hz in frequency, once daily, 10 min each time, for 3, 7 and 14 days successively. Separately, on the day 3, 7 and 14 of intervention, the modified neurological severity scale (mNSS) was used to evaluate the degree of neurological function injury in the rats, HE staining and Nissl staining were to observe the pathological and morphological changes in brain tissue, TUNEL method was to observe the level of apoptosis in brain tissue and immunohistochemistry (IHC) method and Western blot were to determine the protein expressions of Cyt-C and Caspase-9 in brain tissue.@*RESULTS@#Compared with the sham-operation group, on the day 3, 7 and 14 of intervention, mNSS scores were increased obviously in the rats of the model group respectively (<0.01). Compared with the model group, on the day 3, 7 and 14 of intervention, mNSS scores were reduced in the rats of the EA group respectively (<0.05). On day 3 of intervention, in brain injury region of the rats in the model group and the EA group, gross tissue necrosis, nuclear fragmentation, consolidation and obvious vacuolar changes, reduced Nissl bodies and scattered arrangement were found. On day 7 and 14 of intervention, in the model group and the EA group, the new connective tissue filling and normal cells were visible and Nissl bodies increased. The overall repair and Nissl body quantity in the EA group were better than the model group. Compared with the sham-operation group, on day 3, 7 and 14 of intervention, the numbers of apoptotic cells were increased obviously in the model group (<0.01) and they were reduced in the EA group as compared with the model group (<0.05). Compared with the sham-operation group, on day 3, 7 and 14 of intervention, the protein expressions of Cyt-C and Caspase-9 in damaged brain tissue were all increased obviously in the model group (<0.01) and they were all reduced in the EA group as compared with the model group successively (<0.05).@*CONCLUSION@#Electroacupuncture remarkably improves the condition in the neurological function injury and reduces apoptosis degree in TBI model rats, which is likely related to the down-regulation of the protein expressions of Cyt-C and Caspase-9 in damaged brain tissue and further to bring the impacts on mitochondria mediated apoptosis process.


Subject(s)
Animals , Apoptosis , Brain Injuries, Traumatic , Therapeutics , Caspase 9 , Metabolism , Cytochromes c , Metabolism , Electroacupuncture , Random Allocation , Rats , Rats, Sprague-Dawley
4.
Article in English | WPRIM | ID: wpr-764986

ABSTRACT

We analyzed Clonorchis sinensis ancient DNA (aDNA) acquired from the specimens of the Joseon mummies. The target regions were cytochrome C oxidase subunit 1 (CO1), internal transcribed spacer 1 (ITS1), nicotinamide adenine dinucleotide hydrogen (NADH) dehydrogenase subunits 2 (NAD2) and 5 (NAD5). The sequences of C. sinensis aDNA was completely or almost identical to modern C. sinensis sequences in GenBank. We also found that ITS1, NAD2 and NAD5 could be good markers for molecular diagnosis between C. sinensis and the other trematode parasite species. The current result could improve our knowledge about genetic history of C. sinensis.


Subject(s)
Clonorchis sinensis , Cytochromes c , Cytochromes , Databases, Nucleic Acid , Diagnosis , DNA , Electron Transport Complex IV , Hydrogen , Mummies , NAD , Niacinamide , Oxidoreductases , Parasites , Republic of Korea
5.
Article in Korean | WPRIM | ID: wpr-764708

ABSTRACT

OBJECTIVES: To investigate whether the cytotoxic effect of Cimicifuga rhizoma extract is associated with cell death in the human keratinocyte (HaCaT) and human melanoma cell lines (G361). METHODS: Apoptosis induced by Cimicifuga rhizoma extract was confirmed by water-soluble tetrazolium salts-1 (WST-1) assay, immunocytochemistry, and western blot. Additionally, the release of cytochrome c and apoptosis-inducing factor (AIF) was visualized by confocal laser scanning microscopy. RESULTS: The results showed that Cimicifuga rhizoma extract significantly reduced the viability of G361 cells with half-maximal inhibitory concentration (IC 50) of 200 µg/ml, and the apoptotic process was found to occur via the activation of caspase-3 and caspase-9 pathways. Besides, the release of cytochrome c and AIF was also detected. CONCLUSIONS: This study suggests that Cimicifuga rhizoma extract causes apoptosis of human melanoma cells through the intrinsic apoptotic pathway.


Subject(s)
Apoptosis Inducing Factor , Apoptosis , Blotting, Western , Caspase 3 , Caspase 9 , Cell Death , Cell Line , Cimicifuga , Cytochromes c , Humans , Immunohistochemistry , Keratinocytes , Melanoma , Microscopy, Confocal
6.
Yonsei Medical Journal ; : 509-516, 2019.
Article in English | WPRIM | ID: wpr-762085

ABSTRACT

PURPOSE: This study was conducted to verify the induction and mechanism of selective apoptosis in G361 melanoma cells using anti-HER2 antibody-conjugated gold nanoparticles (GNP-HER2). MATERIALS AND METHODS: Following GNP-HER2 treatment of G361 cells, cell cycle arrest and apoptosis were measured by WST-1 assay, Hemacolor staining, Hoechst staining, immunofluorescence staining, fluorescence-activated cell sorting analysis, and Western blotting.


Subject(s)
Actins , Apoptosis Inducing Factor , Apoptosis , Blotting, Western , Caspase 3 , Caspases , Cell Adhesion , Cell Cycle , Cell Cycle Checkpoints , Cell Death , Cyclin A , Cyclin D1 , Cyclin E , Cyclins , Cytochromes c , Cytoplasm , DNA Fragmentation , Down-Regulation , Flow Cytometry , Fluorescent Antibody Technique , Focal Adhesions , Melanoma , Mitochondria , Nanoparticles , Phosphotransferases , ErbB Receptors , Up-Regulation
7.
Article in English | WPRIM | ID: wpr-760353

ABSTRACT

Avian malaria is one of the most important general blood parasites of poultry in Southeast Asia. Plasmodium (P.) juxtanucleare causes avian malaria in wild and domestic fowl. This study aimed to identify and characterize the Plasmodium species infecting in Thai native fowl. Blood samples were collected for microscopic examination, followed by detection of the Plasmodium cox I gene by using PCR. Five of the 10 sampled fowl had the desired 588 base pair amplicons. Sequence analysis of the five amplicons indicated that the nucleotide and amino acid sequences were homologous to each other and were closely related (100% identity) to a P. juxtanucleare strain isolated in Japan (AB250415). Furthermore, the phylogenetic tree of the cox I gene showed that the P. juxtanucleare in this study were grouped together and clustered with the Japan strain. The presence of P. juxtanucleare described in this study is the first report of P. juxtanucleare in the Thai native fowl of Thailand.


Subject(s)
Amino Acid Sequence , Animals , Asia, Southeastern , Asian Continental Ancestry Group , Base Pairing , Cytochromes c , Cytochromes , Electron Transport Complex IV , Humans , Japan , Malaria, Avian , Parasites , Plasmodium , Polymerase Chain Reaction , Poultry , Sequence Analysis , Thailand , Trees
8.
São Paulo; s.n; s.n; 2019. 299 p. tab, graf.
Thesis in English | LILACS | ID: biblio-1023795

ABSTRACT

Protein PEGylation is the covalent bonding of polyethylene glycol (PEG) polymers to amino acid residues of the protein and it is one of the most promising techniques for improving the therapeutic effect of biopharmaceuticals and long-term stability of protein-based biosensors. This chemical modification brings advantages to biopharmaceuticals, such as an increased half-life, enhanced stability, and reduced immunogenicity. Moreover, in the analytical field, PEGylation improves the multiple properties of protein-based biosensors including biocompatibility, thermal and long-term stability, and solubility in organic solvents. However, the use of PEGylated conjugates in the analytical and therapeutic fields has not been widely explored. The limited industrial application of PEGylated bioconjugates can be attributed to the fact that the reaction and separation steps are currently a challenge. The correct selection of the PEGylation reaction design and the purification process are important challenges in the field of bioconjugation. In this sense, the design and optimization of site-specific PEGylation reactions and application of aqueous biphasic systems (ABS) as purification platforms for PEGylated conjugates are the two main objectives of this thesis. Regarding the purification step, the efficient fractionation (i) of the PEGylated conjugates from the native protein and (ii) of the PEGylated conjugates based on their degree of PEGylation was studied. Centrifugal partition chromatography (CPC) was applied as a continuous regime platform based on ABS technology to efficiently purify the PEGylated proteins. The two proteins under study are L-asparaginase, an important biopharmaceutical applied in the treatment of acute lymphoblastic leukemia and cytochrome c, a promising biosensor. The current work developed in this thesis demonstrates the great potential of ABS in the fractionation of PEGylated proteins, under batch and continuous regime. In addition, in situ recovery of the PEGylated products through one-pot bioconjugation and ABS purification was successfully demonstrated for both enzymes studied. Although further research on scale-up is still required, the results presented show the relevance of ABS platforms for the development of separation processes of PEGylated proteins


A PEGuilação de proteínas é a ligação covalente de polímeros de polietilenoglicol (PEG) a resíduos de aminoácidos da proteína e é uma das técnicas mais promissoras para melhorar o efeito terapêutico dos biofármacos e a estabilidade a longo prazo de biossensores proteícos. Esta modificação química traz vantagens aos produtos biofarmacêuticos, como um aumento da meia-vida, maior estabilidade e imunogenicidade reduzida. Além disso, no campo analítico, a PEGuilação melhora as múltiplas propriedades dos biossensores baseados em proteínas, incluindo biocompatibilidade, estabilidade térmica e a longo prazo, e solubilidade em solventes orgânicos. No entanto, o uso de conjugados PEGuilados em campos analíticos e terapêuticos não tem sido amplamente explorado. A aplicação industrial limitada dos bioconjugados PEGuilados pode ser atribuída ao facto de as etapas de reacção e separação serem atualmente um desafio. A seleção correcta do design da reacção de PEGuilação e do processo de purificação são importantes desafios no campo da bioconjugação. Neste sentido, a concepção e otimização de reações de PEGuilação sítio-específicas e aplicação de sistemas aquosos bifásicos (ABS) como plataformas de purificação de conjugados PEGuilados são os dois principais objetivos desta tese. No que concerne à etapa de purificação foi estudado o eficiente fracionamento (i) dos conjugados PEGuilados, da proteína nativa e (ii) dos conjugados PEGuilados baseados no seu grau de PEGuilação. A cromatografia por partição centrífuga (CPC) foi aplicada como uma plataforma de regime contínuo baseada na tecnologia de ABS para purificar eficientemente as proteínas PEGuiladas. As duas proteínas em estudo são a L-asparaginase, importante biofármaco aplicado no tratamento da leucemia linfoblástica aguda e o citocromo c, um potencial biossensor. A partir dos trabalhos desenvolvidos, é possível confirmar o grande potencial dos ABS no fracionamento de proteínas PEGuiladas, em regime contínuo e descontínuo. Além disso, a recuperação in situ dos produtos PEGuilados através da integração em uma única etapa de bioconjugação e purificação por ABS foi comprovada com sucesso para ambas as enzimas estudadas. Embora ainda sejam necessários estudos adicionais sobre a viabilidade destes sistemas em larga escala, os resultados aqui apresentados demonstram a relevância dos ABS para o desenvolvimento de processos de separação de proteínas PEGuiladas


Subject(s)
Polyethylene Glycols/adverse effects , Proteins/analysis , Biological Products/therapeutic use , Proteins/isolation & purification , Cytochromes c
9.
Article in English | WPRIM | ID: wpr-715613

ABSTRACT

Vasicinone, a quinazoline alkaloid from Adhatoda vasica Nees. is well known for its bronchodilator activity. However its antiproliferative activities is yet to be elucidated. Here-in we investigated the anti-proliferative effect of vasicinone and its underlying mechanism against A549 lung carcinoma cells. The A549 cells upon treatment with various doses of vasicinone (10, 30, 50, 70 µM) for 72 h showed significant decrease in cell viability. Vasicinone treatment also showed DNA fragmentation, LDH leakage, and disruption of mitochondrial potential, and lower wound healing ability in A549 cells. The Annexin V/PI staining showed disrupted plasma membrane integrity and permeability of PI in treated cells. Moreover vasicinone treatment also lead to down regulation of Bcl-2, Fas death receptor and up regulation of PARP, BAD and cytochrome c, suggesting the anti-proliferative nature of vasicinone which mediated apoptosis through both Fas death receptors as well as Bcl-2 regulated signaling. Furthermore, our preliminary studies with vasicinone treatment also showed to lower the ROS levels in A549 cells and have potential free radical scavenging (DPPH, Hydroxyl) activity and ferric reducing power in cell free systems. Thus combining all, vasicinone may be used to develop a new therapeutic agent against oxidative stress induced lung cancer.


Subject(s)
Apoptosis , Cell Membrane , Cell Survival , Cell-Free System , Cytochromes c , DNA Fragmentation , Down-Regulation , Justicia , Lung Neoplasms , Lung , Oxidative Stress , Permeability , Receptors, Death Domain , Up-Regulation , Wound Healing
10.
Article in Chinese | WPRIM | ID: wpr-771476

ABSTRACT

OBJECTIVE@#To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes.@*METHODS@#H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells.@*RESULTS@#Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells.@*CONCLUSIONS@#Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.


Subject(s)
Adenosine Triphosphate , Apoptosis , Benzyl Alcohols , Pharmacology , Caspase 3 , Cell Line , Cell Survival , Cyclosporine , Pharmacology , Cytochromes c , Glucosides , Pharmacology , Humans , Hydrogen Peroxide , Pharmacology , Membrane Potential, Mitochondrial , Mitochondrial Membrane Transport Proteins , Physiology , Myocytes, Cardiac , Metabolism , Oxidative Stress , Reactive Oxygen Species
11.
Article in English | WPRIM | ID: wpr-742293

ABSTRACT

Although widely studied, the natural diversity of the hard tick is not well known. In this study, we collected 194 sequences from 67 species, covering 7 genera of hard tick. The 5′ region of the mitochondrial cytochrome c oxidase subunit 1 region (586 bp) has been used to investigate intra- and inter-species variation and the phylogenetic tree of neighbor joining method has been used for assessment. As a result, by comparing the K2P-distance of intra- and interspecies, 30 samples (15.2%) shown that interspecies distance was larger than the minimum interspecfic distance. From the phylogenetic analysis, 86.8% (49) of the species were identified correctly at the genus level. On deeper analysis on these species suggested the possibility of presence cryptic species. Therefore, further work is required to delineate species boundaries and to develop a more complete understanding of hard tick diversity over larger scale.


Subject(s)
Cytochromes c , Cytochromes , Electron Transport Complex IV , Ixodidae , Methods , Trees
12.
Article in English | WPRIM | ID: wpr-714732

ABSTRACT

Because of the unsatisfactory treatment options for breast cancer (BC), there is a need to develop novel therapeutic approaches for this malignancy. One such strategy is chemotherapy using non-toxic dietary substances and botanical products. Studies have shown that Panduratin A (PA) possesses many health benefits, including anti-inflammatory, anti-bacterial, anti-oxidant and anti-cancer activities. In the present study, we provide evidence that PA treatment of MCF-7 BC cells resulted in a time- and dose-dependent inhibition of cell growth with an IC50 of 15 µM and no to little effect on normal human MCF-10A breast cells. To define the mechanism of these anti-proliferative effects of PA, we determined its effect critical molecular events known to regulate the cell cycle and apoptotic machinery. Immunofluorescence and flow cytometric analysis of Annexin V-FITC staining provided evidence for the induction of apoptosis. PA treatment of BC cells resulted in increased activity/expression of mitochondrial cytochrome C, caspases 7, 8 and 9 with a significant increase in the Bax:Bcl-2 ratio, suggesting the involvement of a mitochondrial-dependent apoptotic pathway. Furthermore, cell cycle analysis using flow cytometry showed that PA treatment of cells resulted in G0/G1 arrest in a dose-dependent manner. Immunoblot analysis data revealed that, in MCF-7 cell lines, PA treatment resulted in the dose-dependent (i) induction of p21WAF1/Cip1 and p27Kip1, (ii) downregulation of Cyclin dependent kinase (CDK) 4 and (iii) decrease in cyclin D1. These findings suggest that PA may be an effective therapeutic agent against BC.


Subject(s)
Apoptosis , Breast Neoplasms , Breast , Caspases , Cell Cycle Checkpoints , Cell Cycle , Cell Proliferation , Cyclin D1 , Cyclins , Cytochromes c , Down-Regulation , Drug Therapy , Flow Cytometry , Fluorescent Antibody Technique , Humans , Inhibitory Concentration 50 , Insurance Benefits , MCF-7 Cells , Phosphotransferases
13.
Article in English | WPRIM | ID: wpr-713828

ABSTRACT

BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.


Subject(s)
Apoptosis , Blotting, Western , Breast Neoplasms , Breast , Caspase 3 , Caspase 8 , Caspases, Initiator , Cell Survival , Citrus , Cysteine , Cytochromes c , Cytoplasm , Down-Regulation , Flow Cytometry , Humans , Medicine, Traditional , Membrane Potential, Mitochondrial , Mitochondria , Oxygen , Poly(ADP-ribose) Polymerases , Reactive Oxygen Species , Water
14.
Article in English | WPRIM | ID: wpr-740094

ABSTRACT

BACKGROUND: Prostate cancer (PCa) is one of the most important causes of death in men and thus new therapeutic approaches are needed. In this study, antiproliferative and anti-migration properties of a coumarin derivative esculetin were evaluated. METHODS: Human PCa cell lines PC3, DU145, and LNCaP were treated with various concentrations of esculetin for 24 to 72 hours, and cell viability was determined by the MTT test. Cell cycle and apoptosis were analyzed by using cell-based cytometer. Gene expression levels were assessed by reverse transcription and quantitative real-time PCR, cell migration was determined by the wound healing assay. The protein expression was measured by Western blotting. RESULTS: Esculetin inhibited cell proliferation in a dose- and time-dependent manner. Cell migration was inhibited by esculetin treatment. Administration of esculetin significantly reduced the cells survival, induced apoptosis and caused the G1 phase cell cycle arrest shown by image-based cytometer. The induced expression of cytochrome c, p53, p21 and p27, and down-regulated CDK2 and CDK4 may be the underlying molecular mechanisms of esculetin effect. Esculetin suppressed phosphorylation of Akt and enhanced protein expression of tumor-suppressor phosphatase and tensin homologue. CONCLUSIONS: Our findings showed that the coumarin derivative esculetin could be used in the management of PCa. However, further in vivo research is needed.


Subject(s)
Apoptosis , Blotting, Western , Cause of Death , Cell Cycle Checkpoints , Cell Cycle , Cell Line , Cell Movement , Cell Proliferation , Cell Survival , Cytochromes c , G1 Phase , Gene Expression , Humans , Male , Passive Cutaneous Anaphylaxis , Phosphorylation , Prostate , Prostatic Neoplasms , Real-Time Polymerase Chain Reaction , Reverse Transcription , Wound Healing
15.
São Paulo; s.n; 2018. 201 p.
Thesis in Portuguese | LILACS | ID: biblio-909896

ABSTRACT

Introdução. A identificação dos flebotomíneos baseia-se principalmente na morfologia do adulto, o que pode ser problemático quando as espécies são morfologicamente muito semelhantes. Psychodopygus é um gênero de flebotomíneos de grande interesse em saúde pública devido ao papel de algumas espécies na veiculação de Leishmania spp. no Brasil. No entanto, este gênero inclui espécies com fêmeas morfologicamente indistinguíveis que pertencem à Série Chagasi, sendo elas: P. chagasi, P. complexus, P. squamiventris maripaensis, P. squamiventris squamiventris e P. wellcomei. Objetivos. Investigar a possibilidade de distinguir essas espécies por meio de análises morfométrica e molecular, além de produzir uma distribuição geográfica atualizada para o grupo analisando a probabilidade de ocorrência das espécies através da análise de modelagem de nicho ecológico. Material e Métodos. Foi realizada a análise discriminante na morfometria geométrica (cabeça e asa) e linear, morfologia (usando microscopia óptica e eletrônica de varredura) e a análise do citocromo c oxidase subunidade 1 (COI), avaliando-se um total de 752 espécimes (460 fêmeas e 292 machos) dos seguintes estados Amapá, Amazonas, Ceará, Mato Grosso, Pará, Rondônia, Roraima e Tocantins. Mapas de distribuição foram produzidos através de dados obtidos do material analisado e de revisão bibliográfica. Resultados. A análise discriminante usando caracteres morfométricos lineares mostrou-se capaz de diferenciar todas as espécies, exceto P. complexus, que apresentou 2,2% de erro de identificação. A morfometria geométrica das asas foi incapaz de separar completamente as espécies através da conformação, mas o tamanho do centróide dos espécimes fêmeas falhou apenas em distinguir P. complexus de P. s. maripaensis. Por outro lado, a morfometria geométrica das cabeças foi capaz de distinguir todas as espécies com grande eficiência ao usar tanto a forma como o tamanho do centróide. A análise morfológica revelou que a coloração torácica, principalmente do pronoto e do pós-noto, pode ser usada para separar as cinco espécies em três grupos: P. chagasi, P. wellcomei / P. complexus e P. s. mariapaensis / P. s. squamiventris. Os resultados da análise de DNA Barcoding, mostraram um agrupamento semelhante ao observado na morfologia; embora os espécimes de P. wellcomei do estado do Ceará mostrem uma grande distância genética da população do estado do Pará, evidenciando que essa espécie possa representar um complexo. Quanto à microscopia eletrônica de varredura, foram avaliadas detalhadamente as estruturas das antenas, tórax e genitália masculina. Salientamos que no anepímero (tórax) foi observada uma escama tipo \"raquete\" modificada apenas em Psychodopygus s. squamiventris. A revisão da distribuição geográfica mostrou que as espécies possuem uma distribuição cis-andina, ocorrendo principalmente no bioma Amazônico. A nítida separação de algumas espécies pelo rio Amazonas, sugere que o surgimento do grupo ocorreu no período que se estende da orogênese dos Andes até a formação deste rio. Conclusões. O estudo possibilitou diferenciar completamente as fêmeas das cinco espécies da Série Chagasi utilizando o conjunto de dados obtidos por morfometria linear e geométrica e análises morfológicas e também apresentar novos caracteres morfológicos e padrões distribucionais que facilitarão a identificação de machos e fêmeas dessas espécies


Introduction. The identification of sand flies is mainly based on adult morphology, which can be problematic when species are morphologically very similar. Psychodopygus is one of the sand fly genera of great interest in public health, due to the role of some species in the transmission of Leishmania spp. in Brazil. However, this genus includes species with morphologically indistinguishable females that belong to the Chagasi series, which includes: P. chagasi, P. complexus, P. squamiventris maripaensis, P. squamiventris squamiventris and P. wellcomei. Objectives. To investigate the possibility of distinguishing among these species by means of morphometric and molecular analyses in addition to producing an updated geographical distribution for the group, analyzing the probability of the occurrence of the species by the analysis of ecological niche modeling. Material and methods. The analyses of the cytochrome c oxidase subunit 1 (COI), geometrical (head and wing) and of linear morphometry and morphology (using optical microscopy and scanning electron microscopy) were carried out using a total of 752 specimens (460 females and 292 males) from the following states: Amapá, Amazonas, Ceará, Mato Grosso, Pará, Rondônia, Roraima e Tocantins. Distribution maps were produced on the basis of data obtained from the material analyzed and a bibliographical review. Results. The discriminant analysis using linear morphometric characters was able to differentiate among all the species, except for P. complexus, which presented a 2.2% error of identification. The geometric morphometry of the wings was unable to completely separate the species by means of the shape analyses, but the centroid size of the female specimens only failed to distinguish P. complexus from P. s. maripaensis. Otherwise, the geometric morphometry of the heads was sufficient to distinguish all the species with great efficiency, when using both the head-shape and the centroid size. The morphological analysis revealed that the thoracic coloration, mainly of the pronotum and the post-notum, can be used to separate the five species into three groups: P. chagasi, P. wellcomei / P. complexus, P. s. mariapaensis / P. s. squamiventris. The results of the Barcoding DNA analyses showed a cluster similar to that observed in the morphology; however, P. wellcomei specimens from the Ceará population showed a great genetic distance from the population of Pará, evidencing that this species may represent a complex. As for the scanning electron microscopy, the structures of the antennae, thorax and male genitalia were evaluated in detail. In the anepimerum (thorax) a modified \"racket\"-type scale was observed only in Psychodopygus s. squamiventris. The review of the geographical distribution showed that the species have a cis-Andean distribution, occurring mainly in the Amazonian biome. The separation of some species from the others by the Amazon river suggests that the appearance of the Chagasi series occurred in the period from the orogenesis of the Andes to the formation of this river. Conclusions. The results clearly differentiate the females of the five species of the Chagasi series using the data set of linear and geometric morphometry and morphological analyses, providing new morphological and distributional data that will facilitate the identification of the males and females of this group


Subject(s)
Animals , Female , Animal Distribution , Cytochromes c , Psychodidae/anatomy & histology , Psychodidae/classification , Diptera , Discriminant Analysis , Disease Vectors , Pigmentation
16.
Article in English | WPRIM | ID: wpr-311375

ABSTRACT

<p><b>OBJECTIVE</b>Scorpion (Hemiscorpius lepturus) stings are a public health concern in Iran, particularly in south and southwestern regions of Iran. The gold standard for the treatment of a scorpion sting is anti-venom therapy. However, immunotherapy can have serious side effects, such as anaphylactic shock (which can sometimes even lead to death). The aim of the current study was to demonstrate the protective effect of ozone against toxicity induced by Hemiscorpius lepturus (H. lepturus) venom in mice.</p><p><b>METHODS</b>Eight hours after the injection of ozone to the experimental design groups, the male mice were decapitated and mitochondria were isolated from five different tissues (liver, kidney, heart, brain, and spinal cord) using differential ultracentrifugation. Then, assessment of mitochondrial parameters including mitochondrial reactive oxidative species (ROS) production, mitochondrial membrane potential (MMP), ATP level, and the release of cytochrome c from the mitochondria was performed.</p><p><b>RESULTS</b>Our results showed that H. lepturus venom-induced oxidative stress is related to ROS production and MMP collapse, which is correlated with cytochrome c release and ATP depletion, indicating the predisposition to the cell death signaling.</p><p><b>CONCLUSION</b>In general, ozone therapy in moderate dose can be considered as clinically effective for the treatment of H. lepturus sting as a protective and antioxidant agent.</p>


Subject(s)
Animals , Brain , Metabolism , Cytochromes c , Metabolism , Heart , Kidney , Metabolism , Liver , Metabolism , Male , Membrane Potential, Mitochondrial , Mice , Mice, Inbred BALB C , Muscle, Skeletal , Metabolism , Myocardium , Metabolism , Ozone , Pharmacology , Scorpion Venoms , Toxicity , Scorpions , Physiology , Spinal Cord , Metabolism
17.
Article in English | WPRIM | ID: wpr-287106

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of Huaiqihuang Granules (, HQH), a mixture of Chinese herbs including Trametes robiniophila Murr, Fructus Lycii and Polygonatum sibiricum, on adriamycininduced nephropathy (ADRN) in rats and its underlying mechanisms.</p><p><b>METHODS</b>Rats with ADRN were divided into four groups: the sham group, the model group (distilled water), the low-dose HQH-treated (2 g/kg) group, and the high-dose HQH-treated (4 g/kg) group. Body weight and 24-h urinary protein (Upro) were checked every week. After 5-week intervention, at the end of the study, the rats were sacrificed and blood samples were collected for examination of biochemical parameters, including glomerular morphological makers, podocyte shape, cellular apoptosis, expressions of nephrin, inflammatory and apoptosis markers.</p><p><b>RESULTS</b>HQH ameliorated the rat's general status, proteinuria, renal morphological appearance and glomerulosclerosis. The decreased expression of nephrin in ADRN rats was increased by HQH, as well as the impaired podocyte foot process fusion. Cytosolic levels of p65 and inhibitor of nuclear factor κBα (IκBα) were decreased in ADRN rats, and recovered by the treatment of HQH. Consistently, the induced expression of tumor necrosis factor α (TNF-α), phosphorylated nuclear factor κB p65 (p-NFκB p65) and IκBα in ADRN were markedly suppressed by HQH. In addition, induction of Bax, cleaved caspase-3 and cytochrome C in ADRN rats were suppressed by HQH, indicating the amelioration of apoptosis.</p><p><b>CONCLUSION</b>HQH could ameliorate renal impairments in ADRN rats by increasing nephrin expression, inhibiting NF-κB signaling pathway via the down-regulation of p-NF-κB p65 and p-IκBα, and suppression of glomerular and tubular apoptosis.</p>


Subject(s)
Animals , Apoptosis , Body Weight , Caspase 3 , Metabolism , Chromatography, High Pressure Liquid , Cytochromes c , Metabolism , Doxorubicin , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Kidney , Pathology , Kidney Diseases , Blood , Drug Therapy , Kidney Glomerulus , Pathology , Kidney Tubules , Pathology , Male , Membrane Proteins , Metabolism , NF-KappaB Inhibitor alpha , Metabolism , NF-kappa B , Metabolism , Organ Size , Proteinuria , Blood , Drug Therapy , Rats, Sprague-Dawley , Signal Transduction , Transcription Factor RelA , Metabolism , Tumor Necrosis Factor-alpha , Metabolism , bcl-2-Associated X Protein , Metabolism
18.
Korean Circulation Journal ; : 193-200, 2017.
Article in English | WPRIM | ID: wpr-59345

ABSTRACT

BACKGROUND AND OBJECTIVES: The overall purpose of this study was to investigate the role of cytochrome C oxidase (CcO) in preventing ischemia reperfusion-induced cardiac injury through gaseous signaling molecule pathways. MATERIALS AND METHODS: We used CcO inhibitor, potassium cyanide (KCN) to mimic the pre-treatment of gaseous signaling molecules in a global ischemia/reperfusion (IR) injury model in rats. Intracellular reactive oxygen species (ROS) was determined by measuring mitochondrial H2O2 and mitochondrial complex activity. RESULTS: KCN pre-treatment led to decreased infarction area after IR injury and improved cardiac function. KCN pre-treated group challenged with IR injury was associated with reduced ROS production through inhibition of activity and not downregulation of CcO expression. In addition, KCN pre-treatment was associated with enhanced expression and activity of mitochondrial antioxidase, suggesting the role of CcO in regulating IR injury through oxidative stress. CONCLUSION: KCN pre-treatment reduced the severity of IR injury. The potential mechanism could be increased endogenous anti-oxidase activity and consequently, the enhanced clearance of ROS.


Subject(s)
Animals , Cytochromes c , Cytochromes , Down-Regulation , Electron Transport Complex IV , Infarction , Ischemia , Mitochondria , Myocardial Infarction , Oxidative Stress , Potassium Cyanide , Rats , Reactive Oxygen Species , Reperfusion Injury
19.
Article in English | WPRIM | ID: wpr-32623

ABSTRACT

Doxorubicin (DOX) is a highly effective chemotherapeutic agent; however, the dose-dependent cardiotoxicity associated with DOX significantly limits its clinical application. In the present study, we investigated whether Rb1 could prevent DOX-induced apoptosis in H9C2 cells via aryl hydrocarbon receptor (AhR). H9C2 cells were treated with various concentrations (−μM) of Rb1. AhR, CYP1A protein and mRNA expression were quantified with Western blot and real-time PCR analyses. We also evaluated the expression levels of caspase-3 to assess the anti-apoptotic effects of Rb1. Our results showed that Rb1 attenuated DOX-induced cardiomyocytes injury and apoptosis and reduced caspase-3 and caspase-8, but not caspase-9 activity in DOX-treated H9C2 cells. Meanwhile, pre-treatment with Rb1 decreased the expression of caspase-3 and PARP in the protein levels, with no effects on cytochrome c, Bax, and Bcl-2 in DOX-stimulated cells. Rb1 markedly decreased the CYP1A1 and CYP1A2 expression induced by DOX. Furthermore, transfection with AhR siRNA or pre-treatment with AhR antagonist CH-223191 significantly inhibited the ability of Rb1 to decrease the induction of CYP1A, as well as caspase-3 protein levels following stimulation with DOX. In conclusion, these findings indicate that AhR plays an important role in the protection of Ginsenoside Rb1 against DOX-triggered apoptosis of H9C2 cells.


Subject(s)
Apoptosis , Blotting, Western , Cardiotoxicity , Caspase 3 , Caspase 8 , Caspase 9 , Cytochrome P-450 CYP1A1 , Cytochrome P-450 CYP1A2 , Cytochromes c , Doxorubicin , Myocytes, Cardiac , Real-Time Polymerase Chain Reaction , Receptors, Aryl Hydrocarbon , RNA, Messenger , RNA, Small Interfering , Transfection
20.
Article in English | WPRIM | ID: wpr-812052

ABSTRACT

The aim of the present study was to investigate the protective effects and underlying mechanisms of Garcinia xanthochymus, a perennial medicinal plant native to Yunnan, China, against HO-induced oxidative damage in rat pheochromacytoma PC12 cells. Preincubation of PC12 cells with fruit EtOAc fraction (fruit-EFr., 12.5-50 µmol·L) of G. xanthochymus for 24 h prior to HO exposure markedly improved cell viability and increased the activities of antioxidant enzymes (superoxide dismutase, catalase, and heme oxygenase-1 [HO-1]), prevented lactate dehydrogenase release and lipid peroxidation malondialdehyde production, attenuated the decrease of matrix metalloproteinases (MMP), and scavenged reactive oxygen species (ROS). Fruit-EFr. also reduced BAX and cytochrome C expression and improved BCL-2 expression, thereby decreasing the ratio of BAX to BCL-2. Fruit-EFr. activated the nuclear translocation of NRF2 to increase HO-1 and induced the phosphorylation of AKT. Its cytoprotective effect was abolished by LY294002, a specific inhibitor of PI3K. Taken together, the above findings suggested that fruit-EFr.of G. xanthochymus could enhance cellular antioxidant defense capacity, at least in part, through upregulating HO-1 expression and activating the PI3K/AKT pathway and that it could suppress HO-induced oxidative damage via PI3K/AKT and NRF2/HO-1 signaling pathways.


Subject(s)
Animals , Antioxidants , Metabolism , Pharmacology , Apoptosis , Biological Transport , Cell Survival , Cytochromes c , Metabolism , Fruit , Garcinia , Heme Oxygenase-1 , Metabolism , Hydrogen Peroxide , NF-E2-Related Factor 2 , Metabolism , Oxidative Stress , PC12 Cells , Phosphatidylinositol 3-Kinase , Metabolism , Phosphatidylinositol 3-Kinases , Phosphorylation , Plant Extracts , Pharmacology , Protective Agents , Pharmacology , Proto-Oncogene Proteins c-akt , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats , Signal Transduction , bcl-2-Associated X Protein , Metabolism
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