• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.1-8
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• 2019

The main goal of this study was to determine the genetic diversity among 36 grape cultivars grown in Palestine by using ISSR-polymerase chain reaction (PCR) fingerprints. Among the tested primers, 17 produced reasonable amplification products with high intensity and pattern stability. A total of 57 DNA fragments (loci) separated by electrophoresis on agarose gels were detected and they ranged in size, from 150 to 900 bp. Out of these fragments, 55 (88%) were polymorphic and 2 (3.5%) monomorphic. Our results also revealed an average of 3.1 loci per primer. A minimum of 1 and maximum of 10 DNA fragments were obtained (S-17, #820 and #841) and (S-31) primers, respectively. Therefore, the later primer (S-31) is considered to be the most powerful primer among the tested ones. The genetic distance matrix showed an average distance range of between 0.05 and 0.76. The maximum genetic distance value of 0.76 (24% similarity) was exhibited between the (Shami and Marawi.Hamadani.Adi) as well as (Bairuti and Marawi.Hamadani.Adi) genotypes. On the other hand, the lowest genetic distance of 0.05 (95% similarity) was exhibited between (Jandali.Tawel.Mofarad and Jandali. Kurawi.Mlzlz) along with (Shami.Aswad and Shami.mtartash. mlwn) genotypes. Furthermore, the UPGMA dendrogram generally clusters the grape cultivars into eight major clusters in addition to an isolated genotype. Based on these figures, the cultivars tested in this study could be characterized by large divergence at the DNA level. This is taking the assumption that our region has a very rich and varied clonal grape genetic structure.

• The Plant Pathology Journal
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• v.39 no.5
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• pp.486-493
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• 2023

Cowpea mild mottle virus (CPMMV) is a global plant virus that poses a threat to the production and quality of legume crops. Early and accurate diagnosis is essential for effective managing CPMMV outbreaks. With the advancement in isothermal recombinase polymerase amplification and lateral flow strips technologies, more rapid and sensitive methods have become available for detecting this pathogen. In this study, we have developed a reverse transcription recombinase polymerase amplification combined with lateral flow strips (RT-RPA-LFS) method for the detection of CPMMV, specifically targeting the CPMMV coat protein (CP) gene. The RT-RPA-LFS assay only requires 20 min at 40℃ and demonstrates high specificity. Its detection limit was 10 copies/µl, which is approximately up to 100 times more sensitive than RT-PCR on agarose gel electrophoresis. The developed RT-RPA-LFS method offers a rapid, convenient, and sensitive approach for field detection of CPMMV, which contribute to controlling the spread of the virus.

• Restorative Dentistry and Endodontics
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• v.46 no.2
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• pp.16.1-16.11
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• 2021

Objectives: The aim of this study was to evaluate the shaping ability of the TruShape and Reciproc Blue systems and the apical extrusion of debris after root canal instrumentation. The ProTaper Universal system was used as a reference for comparison. Materials and Methods: Thirty-three mandibular premolars with a single canal were scanned using micro-computed tomography and were matched into 3 groups (n = 11) according to the instrumentation system: TruShape, Reciproc Blue and ProTaper Universal. The teeth were accessed and mounted in an apparatus with agarose gel, which simulated apical resistance provided by the periapical tissue and enabled the collection of apically extruded debris. During root canal preparation, 2.5% sodium hypochlorite was used as an irrigant. The samples were scanned again after instrumentation. The percentage of unprepared area, removed dentin, and volume of apically extruded debris were analyzed. The data were analyzed using 1-way analysis of variance and the Tukey test for multiple comparisons at a 5% significance level. Results: No significant differences in the percentage of unprepared area were observed among the systems (p > 0.05). ProTaper Universal presented a higher percentage of dentin removal than the TruShape and Reciproc Blue systems (p < 0.05). The systems produced similar volumes of apically extruded debris (p > 0.05). Conclusions: All systems caused apically extruded debris, without any significant differences among them. TruShape, Reciproc Blue, and ProTaper Universal presented similar percentages of unprepared area after root canal instrumentation; however, ProTaper Universal was associated with higher dentin removal than the other systems.

• Analytical Science and Technology
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• v.21 no.1
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• pp.48-52
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• 2008

A new forensic saliva test method using $SALIgAE^{(R)}$ was evaluated in this study. The sensitivity and specificity of $SALIgAE^{(R)}$ were examined and compared to those of other saliva test methods such as agarose gel diffusion method and $Phadebas^{(R)}$ test sheet method. $SALIgAE^{(R)}$ showed high sensitivity and specificity to human saliva in addition to quickness. Moreover modified $SALIgAE^{(R)}$ method was cheap and easy to use in crime scene and DNA laboratory. $SALIgAE^{(R)}$ was very stable at room temperature and had no effect on STR typing.

• Journal of Life Science
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• v.11 no.2
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• pp.111-115
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• 2001

This study was designed to clone the SNF2/SW12 helicase-related genes from the fission yeast Schizosaccha-romyces pombe and thereafter to elucidate the common functions of the proteins in this family. The $hrp^{2+}$gene was cloned by polymerase chain reaction amplification using degenerative primers from conserved SNF2 motifs within the ERCC6 gene, which encodes a protein involved in DNA excision repair. Like other SNF2/SW12 family proteins, the deduced amino acid sequence of Hrp2 contains DNA-dependent ATPase/7 helicase domains as well as the chromodomain and the DNA binding domain. This configuration is similar to that of mCHD1 (mouse chromo-ATPase/helicase-DNA-dinding protein 1), suggesting that Hrp2 is a S. pombe homolog of mCHD1, which is thought to function in altering the chromatin structure to control the gene expression. To characterize the function of Hrp2, 4 Uracil-Hrp2 fusion protein, it was purified near homogeneity by affinity chromatography on $Ni^{2+}$-NTA agarose, DEAE-Sepharose ion exchange arid Sephacryl S-200 gel filtration chromatographies. The purified fusion protein exhibited DNA-dependent ATPase activity, which was stimulated by both double-stranded and single-stranded DNA. To determine the steady-state level of $hrp^{2+}$ transcripts during growth, cells were cultured in medium and collected at every 2hr to prepare total RNAs. The northern blot analysis showed that the level of $hrp^{2+}$ transcripts reached its maximum before the cells entered the exponential growth phase and then decreased gradually, This result implies that Hrp2 may be required at early stages of cell growth.h.

• Korean Journal of Veterinary Service
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• v.47 no.2
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• pp.61-72
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• 2024

This study aimed to evaluate the feasibility of percutaneous endoscopic foraminotomy (PEF) for the treatment of intervertebral disc herniation of the thoracolumbar spine in large-breed dogs by comparing it with open hemilaminectomy (OH). Six large-breed canine cadavers were used in the present study. A barium and agarose mixture (BA-gel) simulating intervertebral disc herniation was injected into the spinal canal at two intervertebral spaces (T12-T13, L2-L3) of the thoracolumbar spine in each cadaver. PEF and OH were randomly allocated to the sites in each cadaver. Computed tomography was performed pre- and postoperatively. The incision length, vertebral window size, procedure time, and amount of simulated disc material removed were recorded to compare PEF and OH. Both procedures clearly exposed the simulated disc material and spinal cord. The size of the incision and vertebral window created after PEF was much smaller than those after OH. The surgical duration of PEF was longer than that of OH. However, no significant difference (P>0.05) was observed in the amount of BA-gel removed between PEF and OH. Thus, PEF could be used as an effective surgical option for intervertebral disc herniation of the thoracolumbar region in large-breed dogs in that it could lead to less tissue damage as well as sufficient removal of the simulated disc material.

• Clinical and Experimental Pediatrics
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• v.50 no.1
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• pp.28-32
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• 2007

Purpose : Human angiotensin converting enzyme (ACE) gene shows an insertion/deletion polymorphism in 16 intron, and three genotypes are determined by whether a 287 bp fragment of the DNA is present or not; II, ID and DD genotype. DD genotype has been suggested as a risk factor of chronic nephrotic disease such as IgA nephropathy and diabetic nephropathy, various cardiovascular diseases and several other diseases. ACE activity increases in acute hepatitis, chronic persistent hepatitis, chronic active hepatitis and cirrhosis. On the other hand, patients with fatty livers have normal ACE activity. This study was designed to find out the relation between polymorphsims of the ACE genes and neonatal hyperbilirubinemia in Koreans. Methods : The genomic DNA was isolated from 110 full-term Korean neonates who had hyperbilirubinemia with no obvious causes (serum bilirubin$${\geq_-}12mg/dL$$) and 164 neonates of a control population (serum bilirubin <12 mg/dL). We performed polymerase chain reaction (PCR) to see the allele of the ACE gene. Electrophoresis was done in the PCR products in 1.5 percent agarose gel, and then DNA patterns were directly visualized under ethidium bromide staining. Results : ACE genotypes in the hyperbilirubinemia group are as follows; 26.36 percent for II, 53.64 percent for ID, 20.00 percent for DD, 0.532 for I allele and 0.468 for D allele. These distributions were not significantly different from those in the control group; 24.39 percent for II, 51.83 percent for DI, 23.78 percent for DD, 0.503 for I allele and 0.497 for D allele. Conclusion : In this study, ACE gene polymorphism was detected in the neonatal hyperbilirubinemia and control group. The most frequent genotype was ID. Our results indicate that the ACE gene polymorphism is not associated with the prevalence of neonatal hyperbilirubinemia in Koreans.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.169-175
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• 2016

Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

• Journal of Applied Biological Chemistry
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• v.54 no.4
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• pp.258-264
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• 2011

The Korean edible mountainous vegetable, byeongpungssam, Cacalia firma Komar. (CFK) is a wild plant found in the intermountain areas in Korea. The aim of this study was to investigate its free radical scavenging activity using 1,1-diphenyl-2-picryl-hydrazyl, 2,2-azinobis(3-ethylbenozothiazoline- 6-sulfonic acid) diammonium salt, ferric reducing/antioxidant power assays, an electron spin resonance spectroscopy. We also examined its protective effect against oxidative DNA damage using agarose electrophoresis of ethanol extract of CFK. The protective activity of the extract against the DNA damage induced by HO${\cdot}$ radicals was compared to epicatechin, ascorbic acid and trolox as reference antioxidant compounds. Total phenolic content in the extract was determined spectrometrically according to the Folin-Ciocalteu procedure and calculated as gallic acid equivalents. Total polyphenolic content of the extract was measured in the leaves ($161.53{\pm}1.07{\mu}g/g$) and shoot ($142.45{\pm}0.56{\mu}g/g$). The antioxidant potential of the extracts against some radicals and DNA damage by HO${\cdot}$ radicals showed over 60%, respectively.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.79-88
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• 1998

This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.