scholarly journals ANALYSIS OF EXPRESSION OF MIRNA AND MRNA IN THE CELLULAR MATERIAL OF THE STOMACH LINING OBTAINED BY ESOPHAGOGASTRODUODENOSCOPY IN ORDER TO DETECT DYSPLASIA AND STOMACH CANCER

2021 ◽  
pp. 53-60
Author(s):  
V. V. Anischenko ◽  
A. A. Arkhipova ◽  
S. E. Titov ◽  
T. L. Poloz ◽  
I. V. Bubnov

miRNA and mRNA are highly specific molecular markers, with their own unique expression profile for each type of tissue. They can become a valuable tool in clinical practice in addition to routine esophagogastroduodenoscopy.Purpose. To study the prospects of using a classifier based on miRNA and mRNA profiling in cytological samples for detecting dysplasia and stomach cancer, as well as to compare the obtained data with the results of profiling of 7 miRNAs used for the analysis of both histological and cytological material.Materials and methods. The study included 221 cytological preparations: 108 samples of adenocarcinoma, 27 samples of dysplasia, 86 samples of normal mucosa. The expression level of miRNA-145-5p, -150-5p, -21,-20a, -31-5p, -34a-5p, -375, -125b, -196b, -106b and mRNA of the following genes: TERT, CDKN2A, CKS2, FN1, was determined using real-time reverse transcription polymerase chain reaction. Quantitative comparison of two independent samples for was performed using the Mann-Whitney test. The samples were stratified into different groups using the C5.0 algorithm.Results: when examining cytological samples in cases of miRNA-125b, 145, -196b, -21, -375 and TERT, СKS2, FN1 mRNA, there was achieved a high level of significance of differences in the cancer/norm group; miRNA-375 and FN1 mRNA in the cancer/dysplasia group and miRNA-145, -196b, -20a, and СKS2, TERT mRNA in the norm/dysplasia groups. When comparing the results for the 7 miRNAs that were used to analyze both histological and cytological material, several differences was noted.Conclusion. The practical use of miRNA and mRNA expression (in cytological material data mining technology does not allow differentiating dysplasia and cancer but can help in identifying "high-risk patients" who should repeat esophagogastroduodenoscopy with multifocal forceps biopsy, with mandatory molecular genetic research.

2018 ◽  
Vol 22 ◽  
pp. 102-107
Author(s):  
N. A. Balashenko ◽  
A. A. Semenyak ◽  
A. S. Kornilkova ◽  
I. A. Sen'kevich ◽  
O. V. Prishchepchik ◽  
...  

Aim. Description of the results of molecular genetic research of insects-representatives of the subfamily Cetoniinae (flower chafer). Methods. For the analysis, methods of low-traumatic sampling of material for DNA isolation, polymerase chain reaction method, sequencing of the region of mtDNA COI gene were used. Results. A technique for sampling biological material for isolating DNA from the hind legs of individuals was proposed, which allowed not to remove from the habitat the individuals of the most valuable species, collected on the territory of the Republic of Belarus. A collection of DNA samples of representatives of the subfamily Cetoniinae was prepared. The haplotypes Oxythyrea funesta (Poda, 1761) and Protaetia aeruginosa (Linnaeus, 1767) of representatives of the populations inhabiting the territory of Belarus have been identified. Conclusions. There were significant differences in haplotypes among representatives of the Belarusian and European parts of their areal. Keywords: molecular markers, mtDNA, DNA-barcoding, insects, subfamily Cetoniinae.


2015 ◽  
Vol 2 (2) ◽  
pp. 26-31 ◽  
Author(s):  
A. Paliy ◽  
A. Zavgorodniy ◽  
B. Stegniy ◽  
A. Gerilovych

Due to the absence of elaborated effi cient means for specifi c prevention of bovine tuberculosis, it is ex- tremely important to detect and eliminate the source of infection and to take veterinary and sanitary preven- tive measures. Here the critical role is attributed to disinfection, which breaks the epizootic chain due to the elimination of pathogenic microorganisms in the environment and involves the application of disinfectants of different chemical groups. Aim. To study the tuberculocidal properties of new disinfectants DZPT-2 and FAG against atypical mycobacteria Mycobacterium fortitum and a TB agent Mycobacterium bovis. Methods. The bacteriological and molecular-genetic methods were used. Results. It was determined that DZPT-2 prepara- tion has bactericidal effect on M. fortuitum when used in the concentration of 2.0 % of the active ingredient (AI) when exposed for 5–24 h, while disinfectant FAG has a bactericidal effect in the concentration of 2.0 % when exposed for 24 h. Disinfectant DZPT-2 in the concentration of 2.0 % of the AI, when exposed for 5–24 h, and FAG preparation in the concentration of 2.0 %, when exposed for 24 h, and with the norm of consump- tion rate of 1 cubic decimeter per 1 square meter disinfect the test-objects (batiste, wood, glazed tile, metal, glass), contaminated with the TB agent M. bovis. Conclusions. Disinfecting preparations of DZPT-2 in the concentration of 2.0 % of AI when exposed for 5 h and FAG in the concentration of 2.0 % when exposed for 24 h may be used in the complex of veterinary and sanitary measures to prevent and control TB of farm ani- mals. The possibility of using the polymerase chain reaction as an additional method of estimating tuberculo- cide activity of disinfectants was proven.


2018 ◽  
Vol 4 ◽  
pp. 86-89 ◽  
Author(s):  
Sanjeev Kumar Adhikari ◽  
Arrogya Gyawali ◽  
Sajan Shrestha ◽  
Swoyam Prakash Shrestha ◽  
Meera Prajapati ◽  
...  

A prevalence study was carried to isolate Salmonella typhimurium from blood (n= 50) and gut samples (n=100) of poultry in Kathmandu valley during early 2016. Salmonella typhimurium bacteria isolated in the selective media were biochemically confirmed based on Bergey’s Manual. Two sets of oligonucleotide primers-the genus specific 16S rRNA and the organism specific invA were employed for molecular level confirmation by the Polymerase Chain Reaction (PCR) assay. The amplified fragments in 1% agarose gel observed at 406bp and 285bp, respectively confirmed the isolates to be Salmonella typhimurium. Of 150 samples tested, Salmonella typhimurium were isolated from 49 samples, among which nine were from blood (18%) and forty from the gut (40%). The present result indicated an alarmingly high level of Salmonella typhimurium, which can result inzoonotic infection in humans owing to increased contact with poultry and consumption of poultry products in the Kathmandu valley.


2010 ◽  
Vol 90 (3) ◽  
pp. 357-360 ◽  
Author(s):  
M. Alipanah ◽  
A. Torkamanzehi ◽  
H. Taghavi

Production of bird species such as ostrich (Struthio camelus) has been gaining increasing importance in Iran as well as many other countries. Ostrich, similar to many other species of birds, lacks sexual dimorphism, making it difficult to differentiate between males and females, especially at an early age, which can be problematic in breeding programs. Recently developed molecular genetic methods that utilize polymerase chain reaction (PCR) based techniques can facilitate rapid identification of the bird’s sex in these species using a DNA sample, which can be easily extracted from blood or feather pulps. We successfully applied a PCR-based RFLP technique and sex chromosome primers for sex determination in a sample of 30 Ostrich chicks using DNA extracted from blood and feather pulps. Both DNA samples (blood and feather pulps) provided useful results. However, using feather pulps from 1-day-old chicks can provide an easy and inexpensive method for sex determination in ostrich. Key words: Ostrich (struthio camelus), sex determination, sexual dimorphism, polymerase chain reaction, RFLP


1999 ◽  
Vol 90 (2) ◽  
pp. 348-354 ◽  
Author(s):  
Venita Jay ◽  
Vern Edwards ◽  
Eelco Hoving ◽  
James Rutka ◽  
Laurence Becker ◽  
...  

✓ The results of cytogenetic and molecular genetic analysis of a central neurocytoma are presented. Central neurocytomas are intriguing neoplasms that exhibit primarily neuronal, but also glial characteristics, which indicate an origin from a pluripotential neuroglial precursor. The authors describe an intraventricular neurocytoma in an 11-year-old boy that showed anaplastic features with widespread necrosis and mitoses, as well as extensive calcification and foci that exhibited marked neuronal differentiation with clusters of ganglion cells. Immunohistochemical examination showed prominent synaptophysin and neurofilament positivity and focal glial fibrillary acidic protein positivity. Electron microscopy revealed abundant neuritic processes with microtubules and dense core granules as well as mature ganglion cells. Flow cytometry studies revealed increased S (7.8%) and G2M (9.7%) phase components. Molecular and cytogenetic studies were undertaken to assess whether there were similarities to two other tumor types that exhibit neuronal differentiation, the neuroblastoma and medulloblastoma. Polymerase chain reaction and fluorescence in situ hybridization (FISH) analysis revealed no evidence of amplification of the MYCN oncogene or chromosome 1p deletion, which are common in neuroblastomas. Chromosomal analysis by G banding revealed a complex karyotype, with counts in the near-diploidy range (45–48). Two chromosomes 1 appeared normal on G banding and FISH analysis, with p58 signals present on the distal p arm of both chromosomes 1; however, three additional copies of distal 1q were present in rearrangements with 4 and 7. Although the histological findings indicate a kinship to the neuroblastoma and medulloblastoma, the central neurocytoma appears to have a different karyotypic profile, although more cases need to be assessed using molecular genetic analysis.


1999 ◽  
Vol 10 (10) ◽  
pp. 646-651 ◽  
Author(s):  
T Beattie ◽  
A Moyes ◽  
C Patrizio ◽  
H Young

Tetracycline resistant Neisseria gonorrhoeae (TRNG) contain a 25.2 MDa TetM plasmid encoding a 68KDa cytoplasmic protein which confers high-level tetracycline resistance. The aim of this study was to subtype all TRNG isolated in Scotland between 1992 and 1998. Subtyping was performed by a polymerase chain reaction (PCR) assay which characterizes the TetM plasmid as either the Dutch variant (443 base pair product) or the American variant (777 base pair product). Of the 78 TRNG isolates, 35 were the American variant and 43 were the Dutch variant. TRNG were distributed amongst 30 serovar/auxotype classes, the most common being 1A6/NR (11.5%), 1A6/P (14.1%) and 1B4/NR (14.1%). The country where infection was acquired was known for 36 of the 46 TRNG strains isolated between 1996 and 1998. All infections acquired in Asia and South America were the Dutch variant whereas all infections acquired in Africa were the American variant. A penicillinase plasmid was present in 66% (23/35) of the American variant TRNG compared with 51% (22/43) of the Dutch variant: the 3.2 MDa penicillinase plasmid was found in 87% of the American variant TRNG whereas the 4.4 MDa penicillinase plasmid was found in 68% of the Dutch variant TRNG. We conclude that subtyping of TRNG by PCR is a useful tool in studying the epidemiology of gonococcal infection due to plasmid-mediated resistant isolates.


Author(s):  
O. A. Kryzhanovskaya ◽  
A. V. Lazareva ◽  
I. V. Chebotar ◽  
Yu. A. Bocharova ◽  
N. A. Mayansky

Aim. Characterize spectrum of antibiotics resistance of Acinetobacter baumannii strains, isolated from patients of 8 surgical and reanimation departments of 3 medical institution of Moscow, and determine molecular-genetic mechanisms of stability of their carbapenem-resistant forms. Materials and methods. 95 strains of A. baumannii, isolated from patients of reanimation and surgical departments of Moscow in 2012 - 2014, were studied. Sensitivity of strains to antibiotics was tested phenotypically according to recommendations of EUCAST. The presence ofVIM, IMP, OXA-23, OXA-40, OXA-48, OXA-58 and NDM genes in the studied strains was determined by polymerase chain reaction in real time. Results. 86.3% of strains turned out to be non-sensitive to carbapenems, sensitive - 13.7%. 80.0% of strains were non-sensitive to gentamicin, 80.0% of strains - to netilmicin, 94.7% of strains - to ciprofloxacin, 2.1% - to colistin. 91.6% of isolates have shown non-sensitivity to members of 2 and more classes of antibiotics, 78.9% of strains - to members of 3 classes. 2 strains were panresistant, 4.2% (4/95) of the isolates were sensitive to all the classes of antibiotics. Metallo-P-lactamases were not detected. Genes of carbapenemases (OXA-23 and/or OXA-40) were detected in 85.3% (81/95) of strains, characterized phenotypically as non-sensitive to carbapenems. Conclusion. The results obtained shown an increase of resistance to carbapenems and multiple resistance in clinically significant strains of A. baumannii. Resistance to carbapenems is associated with OXA-23 and OXA-40 genes. The conclusions allow to justify perspectives of introduction of technologies of molecular-genetic testing of antibiotics resistance.


Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 460-466
Author(s):  
GB Lim ◽  
K Jeyaseelan ◽  
EM Wintour

We have used competitive reverse transcription and polymerase chain reaction (RT/PCR) to compare the levels of erythropoietin (Epo) mRNA in the liver and kidneys of the sheep fetus at 60, 80, 100, 130, and 140 days of gestation (term = 145 to 150 days). The effect of dexamethasone infusion in the ewe on Epo gene expression in the 60-day fetus was also investigated. Epo mRNA levels were highest at 60 days of gestation, the earliest age studied, in both liver and kidney. In the liver, Epo mRNA expression declined as gestation proceeded. Kidney Epo mRNA was maintained at a high level until 100 days of gestation, declining significantly in the 130-day fetus (P < .01). Treatment of ewes carrying 60-day fetuses with 0.76 mg/h dexamethasone for 48 hours resulted in a significant decrease in fetal plasma Epo values and Epo mRNA levels in both the liver and kidney. In the dexamethasone-treated fetuses, Epo mRNA in the liver was 52% of control values (P < .05), and in the kidney, 33% of control (P < .001). The results suggest that the kidney may play a more important role as a site of Epo synthesis in the early gestation sheep fetus than previously thought. Glucocorticoids may have a role in the regulation of Epo gene expression.


2018 ◽  
Vol 23 ◽  
pp. 166-169
Author(s):  
V. A. Chekalov ◽  
N. E. Volkova

Aim. Molecular-genetic analysis of the chickpea genotypes for foc0, foc3, foc4 resistance genes to Fusarium oxysporum f. sp ciceris. Methods. Extraction and purification of DNA, spectrophotometry, polymerase chain reaction, electrophoresis in polyacrylamide gels. Results. 35 chickpea lines and varieties of Ukrainian and foreign breeding characterized according to genotyping on foc0, foc3, foc4 genes of resistance to Fusarium oxysporum f. sp ciceris by the microsatellite markers TA59, TR19 and TR59. Fragments of the expected size for all markers were obtained for samples, for which the resistance level was fixed to certain races. Match between data on the presence of a amplification fragment of a certain size and resistance level among other samples is not found. Conclusions. For 35 chickpea varieties and lines the allele state of foc0, foc3, foc4 genes of resistance to the F. oxysporum f. sp ciceris races 0, 3, 4 is established. The variety ‘Pam’yat’ is recommended as a control of resistance to F. oxysporum f. sp ciceris races 0, 3, 4 in the southern Ukraine conditions. Keywords: chickpea, genes, molecular markers, fusarios, resistance.


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