• Title/Summary/Keyword: Gene testing

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Raw Animal Meats as Potential Sources of Clostridium difficile in Al-Jouf, Saudi Arabia

  • Taha, Ahmed E.
    • Food Science of Animal Resources
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    • v.41 no.5
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    • pp.883-893
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    • 2021
  • Clostridium difficile present in feces of food animals may contaminate their meats and act as a potential source of C. difficile infection (CDI) to humans. C. difficile resistance to antibiotics, its production of toxins and spores play major roles in the pathogenesis of CDI. This is the first study to evaluate C. difficile prevalence in retail raw animal meats, its antibiotics susceptibilities and toxigenic activities in Al-Jouf, Saudi Arabia. Totally, 240 meat samples were tested. C. difficile was identified by standard microbiological and biochemical methods. Vitek-2 compact system confirmed C. difficile isolates were 15/240 (6.3%). Toxins A/B were not detected by Xpect C. difficile toxin A/B tests. Although all isolates were susceptible to vancomycin and metronidazole, variable degrees of reduced susceptibilities to moxifloxacin, clindamycin or tetracycline antibiotics were detected by Epsilon tests. C. difficile strains with reduced susceptibility to antibiotics should be investigated. Variability between the worldwide reported C. difficile contamination levels could be due to absence of a gold standard procedure for its isolation. Establishment of a unified testing algorithm for C. difficile detection in food products is definitely essential to evaluate the inter-regional variation in its prevalence on national and international levels. Proper use of antimicrobials during animal husbandry is crucial to control the selective drug pressure on C. difficile strains associated with food animals. Investigating the protective or pathogenic potential of non-toxigenic C. difficile strains and the possibility of gene transfer from certain toxigenic/ antibiotics-resistant to non-toxigenic/antibiotics-sensitive strains, respectively, should be worthy of attention.

Desmopressin responding female nephrogenic diabetes insipidus: a case report

  • Juyeon Lee;Hae Il Cheong;Jung Won Lee;Ki Soo Pai
    • Childhood Kidney Diseases
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    • v.26 no.2
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    • pp.107-110
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    • 2022
  • Nephrogenic diabetes insipidus, decreased ability to concentrate urine, with production of large amounts of urine, is caused by the refractory response of renal tubules to the action of antidiuretic hormone. This rare disorder, known as X-linked nephrogenic diabetes insipidus, is caused by a mutation in the AVPR2 gene. Because it is hereditary, most patients are male. This report highlights a case of nephrogenic diabetes insipidus in a 3-year 5-month-old female; upon presentation to the hospital, her symptoms included frequent urinationand consumptionof a significant amount ofwater,which had begun2 years ago. The results of blood tests showed increased levels of serum antidiuretic hormone, and sellar magnetic resonance imaging showed no abnormality. The results of the water restriction test and the desmopressin administration test confirmed the diagnosis of nephrogenic diabetes insipidus showing a partial response to desmopressin. The results of genetic testing indicated the presence of an AVPR2 mutation, a heterozygous missense mutation (p.Val88Met), suggesting inheritance of X-linked nephrogenic diabetes insipidus. This report describes a significant case of symptomaticX-linked nephrogenic diabetes insipidus in a female patient who showed a partial response to desmopressin.

Protection against spring viremia carp virus (SVCV) by immunization with chimeric snakehead rhabdovirus expressing SVCV G protein

  • Mariem Bessaid;Kyung Min Lee;Jae Young Kim;Ki Hong Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.17-23
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    • 2024
  • Spring viremia of carp virus (SVCV) poses a significant threat to numerous cyprinid fish species, particularly the common carp (Cyprinus carpio), often resulting in substantial mortalities. This study explores the potential use of a chimeric recombinant snakehead rhabdovirus carrying the SVCV G gene (rSHRV-Gsvcv) as a live vaccine against SVCV infection. Through virulence testing in zebrafish at different temperatures (15 ℃ and 20 ℃), no mortality was observed in groups infected with either rSHRV-wild or chimeric rSHRV-Gsvcv at both temperatures, whereas 100% mortality occurred in fish infected with wild-type SVCV. Subsequently, as no mortality was observed by rSHRV-Gsvcv, three independent experiments were conducted to determine the possible usage of chimeric rSHRV-Gsvcv as a vaccine candidate against SVCV infection. Fish were immunized with either rSHRV-Gsvcv or rSHRV-wild, and their survival rates against the SVCV challenge were compared with a control group injected with buffer alone at four weeks post-immunization. The results showed that chimeric rSHRV-Gsvcv induced significantly higher fish survival rates compared to rSHRV-wild and the control groups. These findings suggest that genetically engineered chimeric rSHRV-Gsvcv holds the potential for a prophylactic measure to protect fish against SVCV infection.

Biocontrol of Peach Gummosis by Bacillus velezensis KTA01 and Its Antifungal Mechanism

  • Tae-An Kang;GyuDae Lee;Kihwan Kim;Dongyup Hahn;Jae-Ho Shin;Won-Chan Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.296-305
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    • 2024
  • Peach tree gummosis is a botanical anomaly distinguished by the secretion of dark-brown gum from the shoots of peach trees, and Botryosphaeria dothidea has been identified as one of the fungal species responsible for its occurrence. In South Korea, approximately 80% of gummosis cases are linked to infections caused by B. dothidea. In this study, we isolated microbes from the soil surrounding peach trees exhibiting antifungal activity against B. dothidea. Subsequently, we identified several bacterial strains as potential candidates for a biocontrol agent. Among them, Bacillus velezensis KTA01 displayed the most robust antifungal activity and was therefore selected for further analysis. To investigate the antifungal mechanism of B. velezensis KTA01, we performed tests to assess cell wall degradation and siderophore production. Additionally, we conducted reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis based on whole-genome sequencing to confirm the presence of genes responsible for the biosynthesis of lipopeptide compounds, a well-known characteristic of Bacillus spp., and to compare gene expression levels. Moreover, we extracted lipopeptide compounds using methanol and subjected them to both antifungal activity testing and high-performance liquid chromatography (HPLC) analysis. The experimental findings presented in this study unequivocally demonstrate the promising potential of B. velezensis KTA01 as a biocontrol agent against B. dothidea KACC45481, the pathogen responsible for causing peach tree gummosis.

An approach based on clustering for detecting differentially expressed genes in microarray data analysis

  • Yuki Ando;Asanao Shimokawa
    • Communications for Statistical Applications and Methods
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    • v.31 no.5
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    • pp.571-584
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    • 2024
  • To identify differentially expressed genes (DEGs), researchers use a testing method for each gene. However, microarray data are often characterized by large dimensionality and a small sample size, which lead to problems such as reduced analytical power and increased number of tests. Therefore, we propose a clustering method. In this method, genes with similar expression patterns are clustered, and tests are conducted for each cluster. This method increased the sample size for each test and reduced the number of tests. In this case, we used a nonparametric permutation test in the proposed method because independence between samples cannot be assumed if there is a relationship between genes. We compared the accuracy of the proposed method with that of conventional methods. In the simulations, each method was applied to the data generated under a positive correlation between genes, and the area under the curve, power, and type-one error were calculated. The results show that the proposed method outperforms the conventional method in all cases under the simulated conditions. We also found that when independence between samples cannot be assumed, the non-parametric permutation test controls the type-one error better than the t-test.

MOLECULAR BIOLOGIC ANALYSIS OF FMR-1 GENE TRINUCLEOTIDE REPEATS IN AUTISTIC PATIENTS (자폐장애 환자에서 FMR-1 유전 삼염기 반복의 분자생물학적 분석)

  • Kwak, Ho-Soon;Chun, Hyo-Jin;Chang, Eun-Jin;Kim, Hee-Cheol;Kim, Jung-Bun;Park, Young-Nam;Jung, Chul-Ho
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.11 no.1
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    • pp.3-15
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    • 2000
  • Objectives:There has been a rapid expansion of studies aimed at elucidating the genetic basis of autistic disorder, especially it’ relationship to fragile-X syndrome. The detection of fragile X chromosome(Xq27.3) by cytogenetic analysis has revealed many difficulties in testing. Therefore, to explore the relationship between autistic disorder and fragile X syndrome, this study administered molecular biologic methods which examined an unstable CGG repeat within the fragile X mental retardation-1(FMR-1) gene. Methods:Ninety nine autistic children and eight normal control children were tested. The number of CGG repeats within FMR-1 gene was measured after amplification by PCR, and cytogenetic analysis was also carried out to detect fragile site Xq27.3. Southern blot hybridization, using StB12.3 and/or Pfxa3 probe, was done for the patients showing expansion of more than 50 CGG repeats (premutation). Results:All but two autistic patients had no expansion in CGG repeats by PCR and there was no significant statistical difference in number of CGG repeat in comparison with normal control. Two autistic patients, considered as premutation by PCR analysis, had no full mutation or premutation by Southern blot hybridization. All autistic children tested did not have any abnormal karyotype or fragile site Xq27.3. Conclusions:These results suggest that autistic patients may not have abnormality in FMR-1 gene or abnormal expansion in CGG repeat. In conclusion, fragile X syndrome may not be antecedent of autistic disorder.

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A neonate with hyperornithinemia-hyperammonemia-homocitrullinuria syndrome from a consanguineous Pakistani family

  • Kim, Yoo-Mi;Lim, Han Hyuk;Gang, Mi Hyeon;Lee, Yong Wook;Kim, Sook Za;Kim, Gu-Hwan;Yoo, Han-Wook;Ko, Jung-Min;Chang, Meayoung
    • Journal of Genetic Medicine
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    • v.16 no.2
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    • pp.85-89
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    • 2019
  • Hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome is a rare autosomal recessive urea cycle disorder. HHH is caused by a deficiency of the mitochondrial ornithine transporter protein, which is encoded by the solute carrier family 25, member 15 (SLC25A15) gene. Recently, government supported Korean newborn screening has been expanded to include a tandem mass spectrometry (MS/MS) measurement of ornithine level. We report a case of a neonate with HHH syndrome showing a normal MS/MS measurement of ornithine level. A female newborn was admitted to neonatal intensive unit due to familial history of HHH syndrome. Her parents were consanguineous Parkistani couple. The subject's older sister was diagnosed with HHH syndrome at age of 30 months based on altered mental status and liver dysfunction. Even though the subject displayed normal ammonia and ornithine levels based on MS/MS analysis, a molecular test confirmed the diagnosis of HHH syndrome. At 1 month of age, amino acid analysis of blood and urine showed high levels of ornithine and homocitrulline. After 11 months of follow up, she showed normal growth and development, whereas affected sister showed progressive cognitive impairment despite no further hyperammonemia after protein restriction and standard therapy. Our report is in agreement with a previous Canadian study, which showed that neonatal samples from HHH syndrome patients demonstrate normal ornithine levels despite having known mutations. Considering the delayed rise of ornithine in affected patients, genetic testing, and repetitive metabolic testing is needed to prevent patient loss in high risk patients.

Clinical Features of Staphylococcal Scalded Skin Syndrome Caused by Community-Associated Methicillin-Resistant Staphylococcus aureus in Changwon City, Korea, during 2006 and 2015 (2006년부터 2015년까지 창원 지역에서 발생한 지역사회관련 메티실린내성 황색포도알균에 의한 포도알균 열상 피부증후군의 임상양상)

  • Park, Jun Hyeong;Kim, Min Chae;Kang, Jin Han;Choi, Jae Won;Lee, Hak Sung;Shin, Ju Hwa;Lee, Je Chul;Ma, Sang Hyuk
    • Pediatric Infection and Vaccine
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    • v.26 no.1
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    • pp.42-50
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    • 2019
  • Purpose: We investigated the clinical features and epidemiology of staphylococcal scalded skin syndrome (SSSS) from year 2006 to 2015 in Changwon city, Korea. Methods: We reviewed medical records of 69 patients diagnosed with SSSS from year 2006 to 2015. Antibiotic susceptibility testing was performed by agar dilution method. Methicillinresistant Staphylococcus aureus (MRSA) was phenotypically identified by oxacillin susceptibility testing and genotypically confirmed by the existence of the mecA gene. Results: The median age of patients was 2.0 years (range 0.2-6 years). Three (4.3%), 53 (76.8%), and 13 (18.9%) patients showed the generalized type, the intermediate type, and the abortive type, respectively. Patients occurred throughout the year, but most patients occurred between July and October. MRSA was isolated from 54 of the 60 patients regardless of the clinical types. All patients recovered without any complications. Conclusions: There was a constant occurrence of SSSS patients caused by MRSA in Changwon area during 2006 and 2015. It is needed to constantly monitor the occurrence of patients with SSSS.

Transcriptome sequencing reveals non-coding RNAs respond to porcine reproductive and respiratory syndrome virus and Haemophilus parasuis co-infection in Kele piglets

  • Jing Zhang;Chunping Zhao;Min Yao;Jing Qi;Ya Tan;Kaizhi Shi;Jing Wang;Sixuan Zhou;Zhixin Li
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.663-681
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    • 2024
  • Co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and Haemophilus parasuis (HPS) has severely restricted the healthy development of pig breeding. Exploring disease resistance of non-coding RNAs in pigs co-infected with PRRSV and HPS is therefore critical to complement and elucidate the molecular mechanisms of disease resistance in Kele piglets and to innovate the use of local pig germplasm resources in China. RNA-seq of lungs from Kele piglets with single-infection of PRRSV or HPS and co-infection of both pathogens was performed. Two hundred and twenty-five differentially expressed long non-coding RNAs (DElncRNAs) and 30 DEmicroRNAs (DEmiRNAs) were identified and characterized in the PRRSV and HPS co-infection (PRRSV-HPS) group. Compared with the single-infection groups, 146 unique DElncRNAs, 17 unique DEmiRNAs, and 206 target differentially expressed genes (DEGs) were identified in the PRRSV-HPS group. The expression patterns of 20 DEmiRNAs and DElncRNAs confirmed by real-time quantitative polymerase chain reaction (RT-qPCR) were consistent with those determined by high-throughput sequencing. In the PRRSV-HPS group, the target DEGs were enriched in eight immune Gene Ontology terms relating to two unique DEmiRNAs and 16 DElncRNAs, and the unique target DEGs participated the host immune response to pathogens infection by affecting 15 immune-related Kyoto Encyclopedia of Genes and Genomes enrichment pathways. Notably, competitive endogenous RNA (ceRNA) networks of different groups were constructed, and the ssc-miR-671-5p miRNA was validated as a potential regulatory factor to regulate DTX4 and AEBP1 genes to achieve innate antiviral effects and inhibit pulmonary fibrosis by dual-luciferase reporter assays. These results provided insight into further study on the molecular mechanisms of resistance to PRRSV and HPS co-infection in Kele piglets.

Relationship between Eradication of Helicobacter pylori Infection and Clarithromycin Resistance (Helicobacter pylori 감염의 치료와 Clarithromycin 내성간의 연관성)

  • Sohn Seung Ghyu;Lee Jong Hwa;Lee Jung Hun;Lee Sang Hee
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.177-182
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    • 2005
  • H. pylori strains were isolated from antral biopsies taken during upper endoscopy in 114 dyspeptic patients with no previous therapy against H. pylori. Rapid urease test, PCR amplification of SSA and cagA gene for H. pylori detection, and Western blot for CagA expression detection were performed. H. pylori infected patients were treated with omeprazole, clarithromycin (a macrolide), and amoxicillin. At 6 weeks after the discontinuation of therapy, the bacterial eradication rate was determined by endoscopy. The resistance rate to clarithromycin and amoxicillin was $20.2\%$ and $0.0\%$, respectively. The clarithromycin resistance was mainly caused by the A2142G mutation in the 23S rRNA gene of H. pylori. MICs of clarithromycin for the A2142G mutant isolates were significantly higher than MICs for the A2143G mutant isolates. H. pylori eradication was obtained in all patients with clarithromycin-susceptible isolates but not in patients with clarithromycin-resistant isolates (P = 0.0001). These results did not appear to be biased by any differences in CagA expression. The resistance of H. pylori to clarithromycin included in the therapeutic regimens is the most important reason for treatment failure. H. pylori antimicrobial susceptibility testing of the gastric biopsy culture should be performed before choosing the first triple therapy in infected patients and the increase in prevalence of clarithromycin resistance in Korea was problematic.