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: Mitochondrial DNA (mtDNA) mutations may regulate the progression and chemosensitivity of leukemia. Few studies regarding mitochondrial aberrations and haplogroups in acute myeloid leukemia (AML) and their clinical impacts have been reported. Therefore, we focused on the mtDNA length heteroplasmies minisatellite instability (MSI), copy number alterations, and distribution of mitochondrial haplogroups in Korean patients with AML. METHODS: This study investigated 74 adult patients with AML and 70 controls to evaluate mtDNA sequence alterations, MSI, mtDNA copy number, haplogroups, and their clinical implications. The hypervariable (HV) control regions (HV1 and HV2), tRNA(leu1)gene, and cytochrome b gene of mtDNA were analyzed. Two mtDNA minisatellite markers, 16189 poly-C (¹⁶¹⁸⁴CCCCCTCCCC¹⁶¹⁹³, 5CT4C) and 303 poly-C (³⁰³CCCCCCCTCCCCC³¹⁵, 7CT5C), were used to examine the mtDNA MSI. RESULTS: In AML, most mtDNA sequence variants were single nucleotide substitutions, but there were no significant differences compared to those in controls. The number of mtMSI patterns increased in AML. The mean mtDNA copy number of AML patients increased approximately 9-fold compared to that of controls (P < 0.0001). Haplogroup D4 was found in AML with a higher frequency compared to that in controls (31.0% vs. 15.7%, P=0.046). None of the aforementioned factors showed significant impacts on the outcomes. CONCLUSION: AML cells disclosed more heterogeneous patterns with the mtMSI markers and had increased mtDNA copy numbers. These findings implicate mitochondrial genome instability in primary AML cells. Therefore, mtDNA haplogroup D4 might be associated with AML risk among Koreans.
Adult ; Cytochromes b ; DNA, Mitochondrial ; Genome, Mitochondrial* ; Humans ; Leukemia ; Leukemia, Myeloid, Acute* ; Minisatellite Repeats

In 2017, small, elliptical, brownish purple spots on spears and ferns of asparagus were found in fields of Gangwon-do. The isolated fungal species was identified as an ascomycete Stemphylium vesicarium based on morphological characteristics and molecular phylogenic analyses including nucleotide sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and cytochrome b (cytb). A pathogenicity test revealed that S. vesicarium was the causal agent of purple spot disease on asparagus. The occurrence of purple spots caused by S. vesicarium on asparagus is the first report in Korea.
Ascomycota ; Base Sequence ; Cytochromes b ; Ferns ; Gangwon-do ; Korea ; Oxidoreductases ; Virulence

The identification of species primordium has been one of the hot issues in the identification of traditional Chinese medicine. Sea snake is one of the most valuable Chinese medicinal materials in China. In order to understand the origin and varieties of sea snake in the market, we studied the molecular identification of 46 sea snakes by cytochrome B(Cytb). After comparison and manual correction, the sequence length was 582 bp, and the content of A+T(58.9%) was higher than that of G+C(41.1%). There exist 197 variable sites and 179 parsimony-informative sites of the sequence. There are 44 kinds of sequence alignment with consistency equal to 100%, and 2 kinds equal to 96%. A total of 408 Cytb effective sequences were downloaded from GenBank database, with a total of 68 species. Phylogenetic tree of a total of 454 sea snake sequences with the samples in this study were constructed by neighbor-joining trees and Bayesian inference method, respectively, which can identify 42 samples of medicinal materials, while 4 samples can not be identified because of their low node support. The results showed that the species of the sea snake medicine were at least from 2 genera and 5 species, namely, Aipysurus eydouxii, Hydrophis curtus, H. caerulescen, H. curtus, H. ornatus and H. spiralis. This study suggested that the original species of commercial sea snake are very complex and can provide insight into the identification of sea snakes.
Animals ; Bayes Theorem ; China ; Cytochromes b/genetics* ; Hydrophiidae ; Medicine, Chinese Traditional ; Phylogeny

Tigers are one of the most endangered species over the world and protected internationally or locally. However, they are still traded illegally for the Traditional East Asia Medicine or souvenirs / mementos as well as fake products. In the present study, to identify the species of the specimen known as the tiger's genitalia that was sold for mementos in China, two approaches of genetic tools and morphological comparisons were applied. On the basis of the entire sequences of mitochondrial cytochrome b gene (1,140 bp), the result of nucleotide comparisons showed that the specimen examined here is matched with banteng, Bos javanicus (Identities = 1,138/1,140; 99%). In addition, the examination of external morphological characters revealed that the genitalia of the specimen is much more similar to the one of cows, not to tigers. The outcomes highlighted that there are still some fake products of tigers traded in market and molecular genetic tools could help identifying species of wildlife products.
Base Sequence* ; China ; Cytochromes b ; DNA* ; Endangered Species ; Genitalia ; Medicine, East Asian Traditional ; Molecular Biology ; Sequence Analysis, DNA* ; Tigers*

BACKGROUND/AIMS: Balsalazide is an anti-inflammatory drug used in the treatment of inflammatory bowel disease. Balsalazide can reduce inflammatory responses via several mechanisms, including inhibition of nuclear factor-kappaB (NF-kappaB) activity. Parthenolide (PT) inhibits NF-kappaB and exerts promising anticancer effects by promoting apoptosis. The present investigated the antitumor effects of balsalazide, combined with PT, on NF-kappaB in a representative human colorectal carcinoma cell line, HCT116. METHODS: We counted cells and conducted annexin-V assays and cell cycle analysis to measure apoptotic cell death. Western blotting was used investigate the levels of proteins involved in apoptosis. RESULTS: PT and balsalazide produced synergistic anti-proliferative effects and induced apoptotic cell death. The combination of balsalazide and PT markedly suppressed nuclear translocation of the NF-kappaB p65 subunit and the phosphorylation of inhibitor of NF-kappaB. Moreover, PT and balsalazide dramatically enhanced NF-kappaB p65 phosphorylation. Apoptosis, through the mitochondrial pathway, was confirmed by detecting effects on Bcl-2 family members, cytochrome c release, and activation of caspase-3 and -8. CONCLUSIONS: Combination treatment with PT and balsalazide may offer an effective strategy for the induction of apoptosis in HCT116 cells.
Apoptosis ; Blotting, Western ; Caspase 3 ; Cell Cycle ; Cell Death ; Cell Line ; Colorectal Neoplasms* ; Cytochromes c ; HCT116 Cells ; Humans ; Inflammatory Bowel Diseases ; NF-kappa B ; Phosphorylation

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.
Animals ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA Primers ; genetics ; Drug Contamination ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; Sequence Analysis, DNA

BACKGROUND: CM1 (Centrocyte/-blast Marker I) defined by a mAb developed against concanavalin-A activated PBMC, is expressed specifically on a subpopulation of centroblasts and centrocytes of human germinal center (GC) B cells. Burkitt lymphoma (BL) is a tumor consisting of tumor cells with the characteristics of GC B cell. Previously we reported that CM1 ligation with anti-CM1 mAb induced apoptosis in Ramos (IgM(high)) and Raji (IgM(low)) cells. METHODS & RESULTS: In the present study, we observed that CM1 ligation with anti-CM1 mAb induced Fas ligand and Fas expression in Ramos cells, but not in Raji cells. Furthermore, anti-Fas blocking antibody, ZB4, blocked CM1-mediated apoptosis effectively in Ramos cells, but not in Raji cells. Increased mitochondrial membrane permeabilization, which was measured by DiOC6, was observed only in Raji cells. In contrast to no significant change of Bax known as pro-apoptotic protein, anti-apoptotic protein Bcl-2 was significantly decreased in Raji cells. In addition, we observed that CM1 ligation increased release of mitochondrial cytochrome c and upregulated caspase-9 activity in Raji cells. CONCLUSION: These results suggest that apoptosis induced by CM1-ligation is mediated by Fas-Fas ligand interaction in Ramos cells, whereas apoptosis is mediated by down-regulation of Bcl-2 and subsequent decrease of mitochondrial membrane potential in Raji cells.
Apoptosis* ; B-Lymphocytes ; Burkitt Lymphoma ; Caspase 9 ; Cytochromes c ; Down-Regulation* ; Fas Ligand Protein ; Germinal Center ; Humans ; Ligation* ; Membrane Potential, Mitochondrial* ; Mitochondrial Membranes*

Taenia pisiformis is one of the most important parasites of canines and rabbits. T. pisiformis cysticercus (the larval stage) causes severe damage to rabbit breeding, which results in huge economic losses. In this study, the genetic variation of T. pisiformis was determined in Sichuan Province, China. Fragments of the mitochondrial cytochrome b (cytb) (922 bp) gene were amplified in 53 isolates from 8 regions of T. pisiformis. Overall, 12 haplotypes were found in these 53 cytb sequences. Molecular genetic variations showed 98.4% genetic variation derived from intra-region. F(ST) and Nm values suggested that 53 isolates were not genetically differentiated and had low levels of genetic diversity. Neutrality indices of the cytb sequences showed the evolution of T. pisiformis followed a neutral mode. Phylogenetic analysis revealed no correlation between phylogeny and geographic distribution. These findings indicate that 53 isolates of T. pisiformis keep a low genetic variation, which provide useful knowledge for monitoring changes in parasite populations for future control strategies.
Animals ; China ; Cytochromes b/*genetics ; *Genetic Variation ; Helminth Proteins/*genetics ; Humans ; Mitochondria/*genetics ; Molecular Sequence Data ; Phylogeny ; Rabbits ; Taenia/classification/genetics/*isolation & purification ; Taeniasis/*parasitology