• Childhood Kidney Diseases
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• v.4 no.1
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• pp.33-39
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• 2000

Purpose: Our study was designed to investigate the association of MHC Class II (DR, DQ) allele with IgA nephropathy and its significance as a prognostic factor for progression to ESRD Material and Methods: 69 children with IgA nephropathy with normal renal function(serum creatinine $\leq$ 1.5mg/dL) was classified as group A and 70 patients who received renal transplantation due to IgA nephropathy were selected as group B. The HLA-DQB1 and HLA-DRB1 alleles were studied by polymerase chain reaction using sequence specific primers. We have compared the difference in alleles between these two groups and with normal control and also examined any possible effect of the MHC class II genes on the histopathological severity and prognosis of IgAN. Results: Mean age was $8.8{\pm}2.9$ years in group A and $35.0{\pm}15.5$ years in group B. Male to female ratio was 2.8:1 in group A and 2.5:1 in group B. There was a significantly higher frequency of HLA-$DQB1^*03\;and\;DQB1^*05$ in Group B. The frequency of HLA-$DQB1^*0302\;and\;^*05031$ allele had increasing tendency in Group B(P<0.05). HLA-$DRB1^*03\;and\;^*05$ were more common in Group B(P<0.05). HLA-$DRB1^*04$ allele was the most common DR alleles in both group, but there was no statistical significance. There were no significant correlation with MHC class 13 genes on the hjstopathological severity in Group A. Conclusion: In conclusion, $HLA-DQB1^*0302\;and\;HLA-DQB1^*05031 $ allele seemed to be more common in transplanted patients compared to group with normal renal function suggesting that this allele is associated with poor prognosis in IgAN. However larger studies and follow up are required to confirm this due to uncharacterized heterogeneity in etiopathogenesis of IgA nephropathy and possibly one or more than one gene may exert influence in determining susceptibility to the diseases.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2869-2873
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• 2014

Background: Human papillomavirus (HPV) integration within the E2 gene has been proposed as a critical event in cervical carcinogenesis. This study concerned whether HPV16 status and E2 gene intactness are predictive of radiation response in patients with cervical cancer. Materials and Methods: Biopsies of 44 patients with cervical cancer were collected before or after radiotherapy. The presence of HPV16 was assessed by polymerase chain reaction (PCR) using specific primers for the L1 region. E2 disruption was detected by amplifying the entire E2 gene. Results: HPV16 DNA was found in 54.5% of the clinical samples. Overall, 62.5% of the HPV16 positive tumors had integrated viral genome and 37.5% had episomal genome. There was a tendency of increase of HPV16 E2 negative tumors compared with HPV16 L1 ones in advanced stages (75% versus 20% in stage III respectively). Detection of E2 gene appeared influenced by the radiotherapy treatment, as the percentage of samples containing an intact HPV16 E2 was more frequent in pretreated patients compared to radiotherapy treated patients (66.6% versus 20%). The radiation therapy caused an eight-fold [OR= 8; CI=1.22-52.25; p=0.03] increase in the risk of HPV16 genome disruption. The integration status is influenced by the irradiation modalities, interestingly E2 disruption being found widely after radiotherapy treatment (75%) with a total fractioned dose of 50Gy. Conclusions: This study reveals that the status of the viral DNA may be used as a marker to optimize the radiation treatment.

• Tuberculosis and Respiratory Diseases
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• v.58 no.4
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• pp.352-358
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• 2005

Microsatellites are short tandem repeated nucleotide sequences that are present throughout the human genome. Variations in the repeat number or a loss of heterozygosity around the microsatellites have been termed a microsatellite alteration (MA). A MA reflects the genetic instability caused by an impairment in the DNA mismatch repair system and is suggested to be a novel tumorigenic mechanism. A number of studies have reported that MA in the DNA extracted from the plasma occurs at varying frequencies among patients with a non-small cell lung carcinoma (NSCLC). The genomic DNA from 9 subjects with a non-small cell lung cancer (squamous cell cancer 6, adenocarcinoma 2, non-small cell lung cancer1) and 9 age matched non-cancer control subjects (AMC: tuberculosis 3, other inflammatory lung disease 6) and 12 normal control subjects (NC) were extracted from the peripheral blood leukocytes and plasma. Three microsatellite loci were amplified with the primers targeting the Gene Bank sequence D21S1245, D3S1300, and D3S1234. MA in the form of an allelic loss or a band shift was examined with 6% polyacrylamide gel electrophoresis and silver staining. None (0/12) of the NC subjects less than 40 years of age showed a MA in any of the three markers, while 88.9%(8/9) of the AMC above 40 showed a MA in at least one of the three markers (p<0.05). Sixty percent(6/10) of the control subjects with a smoking history showed a MA in one of the three markers, while 9.1%(1/11) of the control subjects without smoking history showed a MA (p<0.05). However, not only did 66.7%(6/9) of lung cancer patients show a MA in at least one of the three markers but so did 88.9%(8/21) of the AMC patients (p>0.05). In conclusion, a MA in the D21S1245, D3S1300, and D3S1234 loci using DNA extracted from the plasma was detected in 66.7% of lung cancer while no MA was found in the young non-smoking control subjects. However, many of the non-cancer control subjects (aged smokers) also showed a MA, which compromised the specificity of the MA analysis as a screening test. Therefore, a further study with a larger sample size will be needed.

• Journal of Microbiology and Biotechnology
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• v.20 no.9
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• pp.1359-1366
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• 2010

A search of the Streptomyces avermitilis genome reveals that its closest homologs are several O-methyltransferases. Among them, one gene (viz., saomt5) was cloned into the pET-15b expression vector by polymerase chain reaction using sequence-specific oligonucleotide primers. Biochemical characterization with the recombinant protein showed that SaOMT5 was S-adenosyl-L-methionine-dependent Omethyltransferase. Several compounds were tested as substrates of SaOMT5. As a result, SaOMT5 catalyzed O-methylation of flavonoids such as 6,7-dihydroxyflavone, 2',3'-dihydroxyflavone, 3',4'-dihydroxyflavone, quercetin, and 7,8-dihydroxyflavone, and phenolic compounds such as caffeic acid and caffeoyl Co-A. These reaction products were analyzed by TLC, HPLC, LC/MS, and NMR spectroscopy. In addition, SaOMT5 could convert phenolic compounds containing ortho-dihydroxy groups into O-methylated compounds, and 6,7-dihydroxyflavone was known to be the best substrate. SaOMT5 converted 6,7-dihydroxyflavone into 6-hydroxy-7-methoxyflavone and 7-hydroxy-6-methoxyflavone, and caffeic acid into ferulic acid and isoferulic acid, respectively. Moreover, SaOMT5 turned out to be a $Mg^{2+}$-dependent OMT, and the effect of $Mg^{2+}$ ion on its activity was five times greater than those of $Ca^{2+}$, $Fe^{2+}$, and $Cu^{2+}$ ions, EDTA, and metal-free medium.

• The Journal of Advanced Prosthodontics
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• v.9 no.2
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• pp.85-92
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• 2017

PURPOSE. The aim of this study was to evaluate the effect of tooth surface pre-treatment steps on shear bond strength, which is essential for understanding the adhesive cementation process. MATERIALS AND METHODS. Shear bond strengths of different cements with various tooth surface treatments (none, etching, priming, or etching and priming) on enamel and dentin of human teeth were measured using the Swiss shear test design. Three adhesives (Permaflo DC, Panavia F 2.0, and Panavia V5) and one self-adhesive cement (Panavia SA plus) were included in this study. The interface of the cement and the tooth surface with the different pre-treatments was analyzed using SEM. pH values of the cements and primers were measured. RESULTS. The highest bond strength values for all cements were achieved with etching and primer on enamel ($25.6{\pm}5.3-32.3{\pm}10.4MPa$). On dentin, etching and priming produced the highest bond strength values for all cements ($8.6{\pm}2.9-11.7{\pm}3.5MPa$) except for Panavia V5, which achieved significantly higher bond strengths when pre-treated with primer only ($15.3{\pm}4.1MPa$). Shear bond strength values were correlated with the micro-retentive surface topography of enamel and the tag length on dentin except for Panavia V5, which revealed the highest bond strength with primer application only without etching, resulting in short but sturdy tags. CONCLUSION. The highest bond strength can be achieved for Panavia F 2.0, Permaflo DC, and Panavia SA plus when the tooth substrate is previously etched and the respective primer is applied. The new cement Panavia V5 displayed low technique-sensitivity and attained significantly higher adhesion of all tested cements to dentin when only primer was applied.

• The Plant Pathology Journal
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• v.33 no.2
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• pp.133-139
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• 2017

Tan spot, caused by the fungus Pyrenophora tritici-repentis, is economically important foliar disease in Latvia, Lithuania, and Romania; however, race structure from Baltic States and Romania is not known. In this study, we performed genotypic and phenotypic race characterization of a large collection of P. tritici-repentis isolates from these countries to determine race structure and utilize this information for better disease management and breeding wheat for tan spot resistance. We characterized 231 single spore isolates from Latvia (n = 15), Lithuania (n = 107), and Romania (n = 109) for Ptr ToxA and Ptr ToxB genes using two genes specific primers. A subset (139) of 231 isolates were further characterized for their race structure by inoculating them individually on tan spot wheat differentials set. Majority (83%) of the 231 isolates amplified Ptr ToxA gene suggesting prevalence of race 1 and 2. Further, phenotypic characterization of 139 isolates also showed wide prevalence of races 1 (68%), 2 (8%), 3 (11%), and 4 (5%) were also identified from Baltic States as well as Romania. Eighteen of the isolates (13%) did not seem to be of any of the eight known races as they lacked Ptr ToxA gene but they behaved like either race 1 or race 2, suggesting possibility of novel toxins in these isolates as their virulence tools.

• The Journal of the Korea Contents Association
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• v.20 no.8
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• pp.674-684
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• 2020

To investigate the differences of bacterial- and archaeal communities depending on kind of wastewater (municipal/livestock) and on treating conditions of basins, sludges were sampled from 10 basins of 3 municipal wastewater treatment plants(WWTP) with A2O and a activated sludge sample from a livestock WWTP. The metagenomic DNAs of the sludge samples were extracted and amplified with primers, 27F/518R for bacteria and Arch519F/A958R for archaea, and pyrosequenced with Roche 454 GS-FLX Titanium. As results, the bacterial communities in basins of municipal WWTPs were quite different from those of livestock WWTP, but within the same municipal WWTP their community structures were similar to each other regardless of different environmental conditions such as O₂. And their archaeal communities resulted from anaerobic·anoxic basins were clustered only within communities originated from the same WWTP. Furthermore Seo-bu WWTP with high bacterial diversity and species richness performed better N/P-removal compared to the orther WWTPs.

• Journal of Life Science
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• v.8 no.4
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• pp.426-430
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• 1998

The molecular relationships have analyzed between the Bombyx mandarina(wild silkworm) and Bombyx mori strains (domesticated silkworm, geographical silkworms). A total of 166 polymorphic RAPD markers amplified from 35 different primers were used to analyze the molecular relationships among thirteen silkworm strains. The genetic similarity coefficient between Bombyx mandarina and Jam305 showed the lowest genetic similarity value with 0.451, Bombyx mandarina and Bibaekjam showed the highest genetic similarity value with 0.958. These strains were classified into Bombyx mandarina(a wild silkworm) and Bombyx mori(twelve domesticated silkworm) groups upon the genetic similary coefficient of 0.55. Further classificient of 0.60; the 1st sub-group (J111, Bibaekjam, $pnd^{ps}$), the 2nd sub-group (Galwon, C18, od yujam, JAM306, C108), the 3rd sub-group(R-hwang, p50), the 4th sub-group(zebra) and the 5th sub-group(JAM305). According to this study, RAPD markers seems to be a valuable tool for molecular relationships and classification among the silkworms.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.2
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• pp.201-206
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• 1994

Down syndrome is one of the major chromosomal anomalies in Korea. To decrease incidence of Down syndrome, antenatal diagnosis is essential. At present, antenatal diagnosis of Down syndrome is done by karyotyping from chorionic villus sampling, amniocentesis, and cordocentsis. All these methods have some problems such as a risk of abortion, a long waiting time, difficulties in sampling, and so on. The aim of study was to confirm that PCR(Polymerase Chain Reaction) using D21S11 primer could be a diagnostic tool for Down syndrome. PCR using D21S11 primers with $^{32}P$ labeling at 5' end was done in 21 cases of DNA from 21 Trisomy and 20 cases of DNA from normal karyotype. PCR product was running for 10 hours on the 6% polyacrylamide gel under 1,000 V or for 8 hours under 1,500 V. After X-ray film exposure, it was read by densitometry. Normal group showed 1: 1 band or single band. 21 Trisomy group showed 1.3-2: 1 band or 2.3 times of density compared to normal single band or 3 bands. This method gave the result within 24 hours. It can be an useful diagnostic tool to detect 21 Trisomy antenatally, especially in late pregnancy, and in preimplantation diagnosis.

• Journal of the korean academy of Pediatric Dentistry
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• v.25 no.2
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• pp.292-303
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• 1998

The arbitrary primer polymerase chain reaction(AP-PCR) and Southern blot restriction fragment length polymorphism(RFLP) were used to genotype the cariogenic pathogen S. mutans in children. Following the morphologic chracteristics of colony on selective medium for S. mutans, total genomic DNA from 155 strains was extracted by conventional methods. Among 155 strains, 143 strains (92.3%) were confirmed S. mutans by PCR with dexA gene and 114 strains were used in this study. Three random sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 2% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI and BamHI, separated on a 0.7 % agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterised 1.6 kilobases (kb) DNA fragment cloned from gtf B gene of S. mutans. The probe was labeled with isotope[$^{32}P-{\alpha}CTP$], and hybridized fragments were detected with intensifying screen. AP-PCR produced 4-8 DNA bands in the 0.25-10 kb regions and distinguished 9, 10 or 12 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 2 hybridization patterns consisting of 1 DNA fragments 450, 500 bp. These results indicate that AP-PCR is more discriminative method for genotyping of S. mutans.