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.
Cytochromes c ; Cytochromes ; Electron Transport Complex IV ; Ixodidae ; Methods ; Trees

Cytochromes c covalently bind their heme prosthetic groups through thioether bonds between the vinyl groups of the heme and the thiols of a CXXCH motif within the protein. In Gram-negative bacteria, this process is catalyzed by the Ccm (cytochrome c maturation) proteins, also called System I. The Ccm proteins are found in the bacterial inner membrane, but some (CcmE, CcmG, CcmH, and CcmI) also have soluble functional domains on the periplasmic face of the membrane. Elucidation of the mechanisms involved in the transport and relay of heme and the apocytochrome from the bacterial cytosol into the periplasm, and their subsequent reaction, has proved challenging due to the fact that most of the proteins involved are membrane-associated, but recent progress in understanding some key components has thrown up some surprises. In this Review, we discuss advances in our understanding of this process arising from a substrate's point of view and from recent structural information about individual components.
Cytochromes c ; chemistry ; metabolism ; Models, Biological

The feasibility of species identification using sequence analysis of the cytochrome B (Cyt B) gene in mitochondrial DNA was investigated. DNA was extracted from nine different animals that could be easily met in our surroundings and Cyt B gene was amplified. Direct sequencing results for the amplified PCR products were compared with each other. Human was also included. Nucleotide sequence of the Cyt B gene for each animals was also compared with the previously known ones registered in nucleotide databases, Genebank. The inter-species sequence variation was high as the percent similarity of each sequences ranged 64.6-83.5%. Compared to this the percent similarity of sequences obtained here were high when compared to the sequences of the same species registered in the database showing relatively low intra-species variation. This data shows that the nucleotide sequences of Cyt B gene in a certain biological materials can be identified at species level. The applicability of this method to the forensic field is also demonstrated by performing a casework; determination of the origin for the placentae which were commercial available as "invogorant". Points about the use of Cyt B gene in forensic field was also reviewed.
Animals ; Base Sequence ; Cytochromes b* ; Cytochromes* ; DNA ; DNA, Mitochondrial ; Humans ; Placenta ; Polymerase Chain Reaction ; Sequence Analysis

The feasibility of species identification using sequence analysis of the cytochrome B (Cyt B) gene in mitochondrial DNA was investigated. DNA was extracted from nine different animals that could be easily met in our surroundings and Cyt B gene was amplified. Direct sequencing results for the amplified PCR products were compared with each other. Human was also included. Nucleotide sequence of the Cyt B gene for each animals was also compared with the previously known ones registered in nucleotide databases, Genebank. The inter-species sequence variation was high as the percent similarity of each sequences ranged 64.6-83.5%. Compared to this the percent similarity of sequences obtained here were high when compared to the sequences of the same species registered in the database showing relatively low intra-species variation. This data shows that the nucleotide sequences of Cyt B gene in a certain biological materials can be identified at species level. The applicability of this method to the forensic field is also demonstrated by performing a casework; determination of the origin for the placentae which were commercial available as "invogorant". Points about the use of Cyt B gene in forensic field was also reviewed.
Animals ; Base Sequence ; Cytochromes b* ; Cytochromes* ; DNA ; DNA, Mitochondrial ; Humans ; Placenta ; Polymerase Chain Reaction ; Sequence Analysis

BACKGROUND: Microinjectors have been used for cell biology and development, and are useful for the study of cellular morphologic changes with response to the external milieu and intracellularly injected molecules. METHODS: This study was performed to confirm the apoptotic changes induced by intracytoplasmic microinjection of cytochrome c (5 mg/mL) to mouse 3T3 fibroblasts with and without pretreatment of Ac-DEVD-CHO (100 mol/mL), and BSA (bovine serum albumin, 5 mg/mL) as a control, and evaluate the usefulness of microinjection as a method to study apoptosis pathways. RESULTS: Mild focal cytoplasmic fragmentation was seen in the cells microinjected with cytochrome c as early as 10 min after the injection. Apoptotic morphology with apoptotic body formation was observed at 60 min after the injection, and then new apoptotic change of the injected cells was not seen. Cytochrome c-injected cells showed about 31% of apoptotic cells of the total injected cells 50-60 min after the injection. BSA-injected cells did not show apoptosis morphology at 50-60 min after the injections. Caspase-3 inhibitor, Ac-DEVD-CHO-treated cells with cytochrome c microinjection exhibited lower apoptosis indices (average apoptosis index; 11.5+/-8.6%) than non-treated cells of the inhibitor (average apoptosis index; 11.5+/-8.6%). CONCLUSIONS: It was observed that intracellular microinjection of cytochromic c induced apoptosis which was inhibited by Ac-DEVD-CHO, although apoptotic cells were so easily detached that further study could not be performed. However it is thought that microinjection should be a method to study apoptosis and signal transduction with the molecular biological techniques currently available.
Animals ; Apoptosis* ; Caspase 3 ; Cytochromes c* ; Cytochromes* ; Cytoplasm ; Fibroblasts* ; Mice* ; Microinjections* ; Serum Albumin ; Signal Transduction

DNA barcoding was recently introduced to molecular identification of forensically important fly species. So, we have analysed the barcode region (687 nucleotides) of mitochondrial cytochrome oxidase subunit I (COI) gene for four species of Muscidae flies collected from Korea. The sequences were aligned and analysed to construct a phylogenetic tree using DNA Star 5.01(DNAStar Inc) and MEGA 3.1 program(Kumar, Tamura, Nei 2004). Intraspecific variation was not noted between M.stabulans individual to each other. Intraspecific variation ranges of other species were 0.1%, 0.1~0.3% and 0.1~0.6% for O.leucostoma, M.angustifrons and M.domestica, respectively. Interspecific percent distance was minimal(9.7~10.0%) between M.stabulans and M.angustifrons. Other species showed above 10% distance from each other. The result showed that four species of Muscidae fly species (Muscina angustifrons, Muscina stabulans, Ophyra leucostoma and Musca domestica) were identifiable from each other with analysis of barcode region of COI gene. Therefore, we conclude that species identification of forensically important Muscidae flies used in this study is possible with percent distance of sequences of COI barcode region, but more species and individuals should be examined to be confident about the conclusion.
Cytochromes* ; Diptera ; DNA* ; Electron Transport Complex IV* ; Korea* ; Muscidae*

Paraquat has been suggested to induce apoptosis by generation of reactive oxygen species (ROS). However, little is known about the mechanism of paraquat-induced apoptosis. Here, we demonstrate that extracellular signal-regulated protein kinase (ERK) is required for paraquat-induced apoptosis in NIH3T3 cells. Paraquat treatment resulted in activation of ERK, and U0126, inhibitors of the MEK/ERK signaling pathway, prevented apoptosis. Moreover, paraquat-induced apoptosis was associated with cytochrome C release, which could be prevented by treatment with the MEK inhibitors. Taken together, our findings suggest that ERK activation plays an active role in mediating paraquat-induced apoptosis of NIH3T3 cells.
Apoptosis* ; Cytochromes c* ; Negotiating ; Paraquat* ; Protein Kinases ; Reactive Oxygen Species

1)The oxygen consumption was studied with albino rate under normal environment after they were given Cytochrome C intravenously (10mg/kg). The consumption was 74.6cc/kg min. with that of control, 75.4cc/kg. min. The difference of the consumptions was not statistically significant. However, under 0.5% CO environment, the oxygen consumption of the Cytochrome C treated rats (62.5cc/kg min) was significantly greater than the control(42.1cc/kg min.) 2) The recovery time of rat acutely poisoned by 1% CO was studied. The recovery time of the Cytochirome C treated group was 37.2 minutes and in control group it was 52.2 minutes. Also significant difference of fatality was noted between the treated group(21.8%) and the untreated group(49.7%). 3) The combined effects of the hyperbaric oxygenation (100% O2 at atmospheric pressures) and the Cytochrome C administration was compared with the effect the simple hyperbaric oxygenation. There was no significant difference of recovery time between the experimental group while the fatality of the experiment group was lower than control group.
Animals ; Cytochromes c ; Hyperbaric Oxygenation* ; Oxygen Consumption ; Poisoning* ; Rats

ABSTRACT: Apoptosis of osteosarcoma cells induced by bile duct derivates, HS-1200 was investigated with relation to mitochodria. HS-1200 induced cytochrome c and Smac/DIABLO release from mitochondria which are major factors related to apoptosis. In these apoptosis processes, release of cytochrome c was not blocked by caspase inhibitor, but release of Smac/DIABLO was blocked. BKA, a kind of PTP (permeablity transition pore) inhibitor, did not block both of them. Interestingly, the alteration of MMP was not observed by means of using JC-1 dye. Although MitoTracker, DiOC-6 and Rhodamine123 were used to confirm previous results, the decrease of MMP was not observed. In order to investigate whether this phenomenon is apoptosis-specific or cell-specific process, genistein was added to cells which usually decreased MMP. After adding genistein, MMP was not decreased, suggesting this phenomenon is cell-specific process. Conclusionally, HS-1200 induced apoptosis of osteosarcoma cells via mitochondria, cytochrome c and Smac/DIABLO were released from mitochondria without decrease of MMP. The release of Smac/DIABLO was dependent of caspase.
Apoptosis* ; Bile Ducts ; Bile* ; Cytochromes c ; Genistein ; Humans* ; Mitochondria ; Osteosarcoma*

This study was undertaken to know whether the UV-irradiation of the skin causes changes in the superoxide dismutase(SOD) activities. After shaving, the back skin of the rabbit was irradiated with UV light ranging from 280 to 320 nm of wavelengths from Burdick lamb (UV800) in doses of either 0.5, 1.0 or l.5 J/cm2. The skin was removed imrnediately after irradiation and the enzyme activity was assayed by the method of McCord ad Fridovich (xanthine-xanthie oxidase system), One unit of the SOD activity was defined as the amount of the enzyme required to inhibit the rate of reduction of cytochrome c by 50%. The protein content of the enzyme was determined by biuret method. The SOD activity of the skin irradiated with 1.0 J/cm was 7. 78 + 1.62 unit/mg protein(Mean+SD; n=10), significantly higher than that of the control(nonirradiated) group(5.62+1.57 unit/mg protein; n=l0). No significant changes were found in the skins irradiated with 1.5 and 1.5 J/cm. This finding indicates that UV irradiation is capablc of increasing the SOD activitiea in the rabbit skin, and suggests that increased superoxide formed by UV irradiatiun induces the increased SOD activies.
Biuret ; Cytochromes c ; Oxidoreductases ; Skin* ; Superoxide Dismutase* ; Superoxides* ; Ultraviolet Rays