Virus isolate G6 was obtained from Hibiscus rosa-sinensis showing yellow and leaf curl symptoms in Guangzhou, Guangdong Province. The complete nucleotide sequence of DNA-A was determined to be 2 737 nucleotides encoding six potential ORFs. Comparison showed that G6 DNA-A had more than 89% sequence identify with all isolates of Cotton leaf curl Multan virus (CLCuMV) and shared the highest sequence identify (96.1%) with CLCuMV isolate 62. G6 DNA-A had 87.1%-89.8% sequence identity with those of CLCuRV isolates, while less than 87% identities with other begomoviruses. Phylogenetic analysis of G6 DNA-A and selected begomoviruses showed that G6 was most closely related to CLCuMV isolates, and they clustered together as a separate branch. Satellite DNA molecule (G6 DNAbeta) was found to be associated with G6 using the primers beta01 and beta02. G6 DNAbeta contains 1346 nucleotides, with a potential functional ORF (C1) in complementary sense DNA. Pairwise comparison indicated that G6 DNAbeta had the highest sequence identities with CLCuMV DNAbeta (92.1%) and CLCuRV DNAbeta (88.7%), but less than 80% sequence identities with other reported satellite DNA molecules. Phylogenetic analysis indicated that G6 DNAbeta was most closely related to CLCuMV DNAbeta and the two DNAbetas clustered together as a separate branch, and formed the main branch with DNAbeta of CLCuRV and MYVV-Y47. It is concluded that G6 infecting Hibiscus rosa-sinensis is an isolate of CLCuMV.
Base Sequence ; DNA, Satellite ; chemistry ; DNA, Viral ; chemistry ; Geminiviridae ; classification ; genetics ; Gossypium ; virology ; Hibiscus ; virology ; Phylogeny

Hypervariable tandem repetitive regions in human DNA are proving to be increasingly useful for genetic analysis in humans. We chose four single locus probes (SLP; MS1, MS43, MS8 and g3) for a validation test among Koreans. The specimens were from 216 unrelated individuals and 33 paternity inclusion families. Extracted DNA from EDTA blood was restricted by Hinfl and electrophoresed in 0.7% agarose gel, transferred and hybridized with chemiluminescent probes. Heterozygosity was over 90% by all of the probes. Total numbers of unassignable mutant bands from 33 paternity inclusion cases were 5, and the highest mutation rate was determined in probe MS1(0.045). The probability of having the same DNA band between two unrelated individuals was 5.7 x 10(-10) when four SLPs were used at the same time. The data presented here on allele frequencies and mutation rates provide preliminary data supporting the validity of these probes in paternity analysis and forensic investigators in the Korean population.
Alleles ; Chromosome Mapping ; DNA, Satellite/*genetics ; Female ; Heterozygote ; Human ; Male ; Mutation ; Myoglobin/*genetics ; *Paternity ; Support, Non-U.S. Gov't

Although the number of protein-coding genes is not highly variable between plant taxa, the DNA content in their genomes is highly variable, by as much as 2,056-fold from a 1C amount of 0.0648 pg to 132.5 pg. The mean 1C-value in plants is 2.4 pg, and genome size expansion/contraction is lineage-specific in plant taxonomy. Transposable element fractions in plant genomes are also variable, as low as ~3% in small genomes and as high as ~85% in large genomes, indicating that genome size is a linear function of transposable element content. Of the 2 classes of transposable elements, the dynamics of class 1 long terminal repeat (LTR) retrotransposons is a major contributor to the 1C value differences among plants. The activity of LTR retrotransposons is under the control of epigenetic suppressing mechanisms. Also, genome-purging mechanisms have been adopted to counter-balance the genome size amplification. With a wealth of information on whole-genome sequences in plant genomes, it was revealed that several genome-purging mechanisms have been employed, depending on plant taxa. Two genera, Lilium and Fritillaria, are known to have large genomes in angiosperms. There were twice times of concerted genome size evolutions in the family Liliaceae during the divergence of the current genera in Liliaceae. In addition to the LTR retrotransposons, non-LTR retrotransposons and satellite DNAs contributed to the huge genomes in the two genera by possible failure of genome counter-balancing mechanisms.
Angiosperms ; Classification ; DNA ; DNA Transposable Elements* ; DNA, Satellite ; Epigenomics ; Fritillaria ; Genome ; Genome Size* ; Genome, Plant ; Humans ; Liliaceae ; Lilium ; Plants ; Retroelements ; Terminal Repeat Sequences

OBJECTIVE: Adeno associated virus (AAV) is a human DNA virus and is included in the Parvovirus family. AAV has been detected in cervical tissues as well as cervical cancer cell lines. Previous studies showed that AAV infection has some negative effects on HPV infection and that the cervical cancer cell growth is inhibited by AAV infection. The aim of this study is to determine the prevalence of AAV 2 infection and its possible roles for influencing HPV 16 and 18 infection in Korean women by analyzing adjacent normal, CIN, and invasive cervical cancer tissue samples. METHODS: CIN I (20), CIN II (24), CIN III (25), invasive cervical cancer (23) tissues were investigated by microdissection and PCR analyses using primers of HPV 16, 18 and AAV 2 as well as beta- globin as an internal control. RESULTS: AAV 2 was detected in 57 out of 92 cervical lesion biopsies. Among these, mild dysplasia, moderate dysplasia, severe dysplasia and invasive cancer showed 55% (11/20), 95.8% (23/24), 52% (13/25) and 52.2% (12/23), respectively. However, HPV 16 was detected in 14 out of 92 cervical lesion biopsies. Among these, mild dysplasia, moderate dysplasia, severe dysplasia and invasive cancer showed 0% (0/20), 8.3% (2/24), 24% (6/25) and 26.1% (6/23), respectively. HPV 18 was detected in 3 out of 92 cervical lesion biopsies. Among these, mild dysplasia, moderate dysplasia, severe dysplasia and invasive cancer showed 0% (0/20), 4.2% (1/24), 8% (2/25) and 0% (0/23), respectively. In contrast, In 92 perilesional normal biopsies, AAV 2, HPV 16 and HPV 18 were detected to be 57.6% (53/92), 3.3% (3/92) and 0% (0/92), respectively. CONCLUSION: AAV 2 was detected in CIN and invasive cervical cancer biopsies by microdissection and PCR analyses in Korean women. It is difficult to confirm any significant roles of AAV 2 infection for developing cervical cancer. However, we observe that there is some correlation between AAV 2 and HPV infection in the carcinogenesis of cervical cancer. Further research remains to be done to further elucidate AAV 2 infection and its role for HPV infection and cervical cancer.
Biopsy* ; Carcinogenesis ; Cell Line ; Dependovirus* ; DNA Viruses ; Female ; Globins ; Human papillomavirus 16 ; Human papillomavirus 18 ; Humans* ; Microdissection* ; Papilloma* ; Parvovirus ; Polymerase Chain Reaction ; Prevalence ; Satellite Viruses ; Uterine Cervical Neoplasms

Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West or al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS) Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at 37degrees C for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at 4degrees C. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol.. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious 'nullisomy' due to hybridization failure without inducing spurious 'disomy' resulting from increased distances between split signals.
Centrifugation ; Chromosomes, Human, Pair 1 ; Dithiothreitol ; DNA, Satellite ; Edetic Acid ; Fluorescence* ; Head ; Humans* ; In Situ Hybridization* ; Semen ; Sex Chromosomes ; Spermatozoa* ; Tissue Donors

OBJECTIVE: Human papillomavirus (HPV) has been identified in the majority of invasive cervical cancer patient and has been found to contribute in a significant way to the genesis of human cervical cancer. HPV has two transforming genes that encode the oncoproteins E6 and E7, E6 can form complexes with p53 and promote p53 degradation, E7 inhibit retinoblastoma protein (RB). The p53 protein is as a phosphoprotein which co-immunoprecipitated with the SV40 T-Antigen. The wild type p53 protein is capable of suppressing the tumorigenic phenotype and regulating cell cycle. Adeno-associated virus(AAV) is a linear single stranded DNA parvovirus which is dependent upon cotransfection by a second unrelated virus to undergo productive infection. It has been well documented that AAV DNA integrates into cellular DNA as one to several tandem copies joined to cellular DNA through the termini. In order to introduce wild type p53 through AAV virus into a cervical cancer patient for gene therapy, we had constructed recombinant p53 adeno associated virus plasmid (pAAVCMVp53). METHODS: pAAVCMVp53 was created new AAV-vector system, pRc/CMVp53 including p53 cDNA and AAV-derivative vector, pASPA-AAV-CMV-polyA were made to HindIII/blunt fragments. Eluated 1.8 kb fragment of wild type p53 cDNA was ligated to pAAV-CMV-polyA, 4.9 kb fragment deprived hASPA cDNA. RESULT: Recombinant AAVCMVp53 was constructed by using pRc/CMVp53 andpASPA-AAV-CMV-polyA. This pAAVCMVp53 was confirmed by various restriction enzyme-digestions and Southern-blotting. This new vector system will be studied on expression, stability in cervical cancer cell lines and animals. CONCLUSION: This system will be one of the useful vector system for cervical cancer gene therapy.
Animals ; Antigens, Viral, Tumor ; Cell Cycle ; Cell Line ; Clone Cells ; DNA ; DNA, Complementary ; DNA, Single-Stranded ; Genetic Therapy* ; Humans ; Oncogene Proteins ; Oncogenes ; Parvovirus ; Phenotype ; Plasmids ; Retinoblastoma Protein ; Satellite Viruses ; Uterine Cervical Neoplasms*

Major aneuploidies diagnosed prenatally involve the autosomes 13, 18, and 21, and sex chromosomes. Fluorescence in situ hybridization (FISH) allows rapid analysis of chromosome copy number in interphase cells. The purpose of this study was to evaluate the role of multicolor fluorescence in situ hybridization in simultaneous detection of probe sets for chromosome 18, X, and Y in uncultured amniotic fluid cells as a safer alternative method for aneuploidy detection prenatally. Fifty amniotic fluid samples were analyzed by FISH and standard cytogenetics. Mean time to obtain results was three days for fluorescence in situ hybridization and 20 days for karyotype. Fluorescence in situ hybridization was informative in 43 samples (86%), and within this group, two aneuploidies were correctly identified. This evaluation demonstrates that FISH with X, Y, and 18 alpha satellite DNA probes could accurately and rapidly detect aneuploidies involving these chromosomes and could be used in any prenatal clinical laboratory.
Amniocentesis/methods* ; Amniotic Fluid/cytology ; Aneuploidy ; Centromere/genetics ; Chromosomes, Human, Pair 18* ; Color ; DNA Probes ; DNA, Satellite/analysis ; Female ; Human ; In Situ Hybridization, Fluorescence/methods* ; Karyotyping ; Pregnancy ; Sex Chromosome Abnormalities/genetics ; Sex Chromosome Abnormalities/diagnosis* ; X Chromosome* ; Y Chromosome*

OBJECTIVESTo investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.

METHODSOne hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.

RESULTS4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).

CONCLUSIONSIn ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.

Adaptor Proteins, Signal Transducing ; Base Pair Mismatch ; Carrier Proteins ; Chromosomal Instability ; Cystadenocarcinoma, Mucinous ; genetics ; DNA Methylation ; DNA Repair ; DNA, Neoplasm ; genetics ; DNA, Satellite ; Female ; Genes, Neoplasm ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics

AIM: Mangrove is one of the oldest living tree species and its leaves are among the most extensively studied botanicals in use today. Scientific research throughout the world has found evidence to support the fact that its foliar extracts have great potential against human microbial pathogens. This study highlights the isolation of foliar fungi from Rhizophora mucronata, Avicenna officialis and Avicenna marina. METHOD: It was isolated in Sabouroud's Dextrose Agar and mass cultivation was done in Sabouroud's Dextrose broth. RESULTS: The ethyl acetate extract showed maximum antibacterial activity which inturn checked for different concentration against bacterial pathogens and anticancer activity for Hep2 and MCF7 cell line in vitro. The DNA was isolated from the fungi and the ITS region of 5.8 s RNA was sequenced and assigned to new species as they are separated from the type strains phylogenetic neighbors by sequence similarities. CONCLUSION This preliminary screening of fungal endophytes revealed their potential to yield potent bioactive compounds for drug discovery programmes.
Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Avicennia ; microbiology ; Base Sequence ; Biological Products ; pharmacology ; therapeutic use ; Cell Line, Tumor ; DNA, Fungal ; Endophytes ; Humans ; Hypocrea ; genetics ; MCF-7 Cells ; Neoplasms ; drug therapy ; Phylogeny ; Phytotherapy ; RNA, Satellite ; Rhizophoraceae ; microbiology ; Species Specificity