scholarly journals FN1 mRNA expression of fibronectin 1 and distribution of fibronectin-associated leukocytes in humans with chronic diffuse liver diseases

2020 ◽  
Vol 11 (2) ◽  
pp. 200-206
Author(s):  
H. V. Dolhikh ◽  
H. S. Maslak ◽  
G. P. Chernenko ◽  
О. H. Minchenko ◽  
А. О. Dolhikh
Keyword(s):  
Polymerase Chain Reaction ◽  
Mrna Expression ◽  
Real Time ◽  
Liver Diseases ◽  
Support Vector ◽  
Control Group ◽  
Chain Reaction ◽  
Prognostic Accuracy ◽  
Polymerase Chain ◽  
Diffuse Liver Diseases

Chronic diffuse liver diseases are characterized by continuous progression of fibrosis, ultimately leading to cirrhosis with the following loss of the normal functioning of this organ due to excessive accumulation of the components of extracellular matrix. To find new, more available diagnostic markers of detecting disorders in the liver, we used methods of antifungal cytofluorometry and quantitative real-time polymerase chain reaction. Intensity of exposure of fibronectin and plasmatic membrane of lymphocytes in the group of patients with chronic diffuse diseases compared with the control group of practically healthy donors decreased both inside and on the surface of the cells respectively by 45.3% and 16.2%. Similar tendency towards decrease was observed during the assays of the level of the exposure of fibronectin on the surface and inside the blood granulocytes: by 25.0% and 36.5%, respectively. In the blood of the patients suffering from chronic diffuse diseases, compared with the control group, there was determined reliable increase in percentage of lymphocytes and granulocytes which contain topical fibronectin, by 32.3% and 2.78 times, correspondingly. The level of monocytes (as a percentage) with cell-associated fibronectin and fibronectin localized inside, by contrast, reliably decreased in 2.07 and 4.50 times, respectively. Analysis of the expression of FN1 in lymphocytes of blood of the studied groups using quantitative real-time polymerase chain reaction revealed decrease in the level of FN1 mRNA expression by 34.0% in the group of ill patients compared with the control group. We determined excellent diagnostic informativeness of the parameters of the level of exposure of fibronectin inside and on the surface of granulocytes and prognostic accuracy of the classifier from these parameters at the level of 100% using the method of support vector machine, SVM. High levels of diagnostic informativeness were recorded for the tests of all types of analyzed leukocytes with cell-associated fibronectin, and the classifiers based on the pair combinations of the tests with cell-associated fibronectin and fibronectin localized within the cells provide high diagnostic accuracy of the prognosis. Because the mentioned indicators are highly-sensitive tests, they can be proposed for early diagnostics and evaluation of the effectiveness of the conducted therapy of chronic diffuse liver diseases, which would allow reducing the use of paracentetic trepanobiopsy, a painful and risky procedure, which still remains the main type of diagnostic.

scholarly journals FN1 mRNA expression of fibronectin 1 and distribution of fibronectin-associated leukocytes in humans with chronic diffuse liver diseases

2020 ◽  
Vol 11 (2) ◽  
pp. 200-206
Author(s):  
H. V. Dolhikh ◽  
H. S. Maslak ◽  
G. P. Chernenko ◽  
О. H. Minchenko ◽  
А. О. Dolhikh
Keyword(s):  
Polymerase Chain Reaction ◽  
Mrna Expression ◽  
Real Time ◽  
Liver Diseases ◽  
Support Vector ◽  
Control Group ◽  
Chain Reaction ◽  
Prognostic Accuracy ◽  
Polymerase Chain ◽  
Diffuse Liver Diseases

Chronic diffuse liver diseases are characterized by continuous progression of fibrosis, ultimately leading to cirrhosis with the following loss of the normal functioning of this organ due to excessive accumulation of the components of extracellular matrix. To find new, more available diagnostic markers of detecting disorders in the liver, we used methods of antifungal cytofluorometry and quantitative real-time polymerase chain reaction. Intensity of exposure of fibronectin and plasmatic membrane of lymphocytes in the group of patients with chronic diffuse diseases compared with the control group of practically healthy donors decreased both inside and on the surface of the cells respectively by 45.3% and 16.2%. Similar tendency towards decrease was observed during the assays of the level of the exposure of fibronectin on the surface and inside the blood granulocytes: by 25.0% and 36.5%, respectively. In the blood of the patients suffering from chronic diffuse diseases, compared with the control group, there was determined reliable increase in percentage of lymphocytes and granulocytes which contain topical fibronectin, by 32.3% and 2.78 times, correspondingly. The level of monocytes (as a percentage) with cell-associated fibronectin and fibronectin localized inside, by contrast, reliably decreased in 2.07 and 4.50 times, respectively. Analysis of the expression of FN1 in lymphocytes of blood of the studied groups using quantitative real-time polymerase chain reaction revealed decrease in the level of FN1 mRNA expression by 34.0% in the group of ill patients compared with the control group. We determined excellent diagnostic informativeness of the parameters of the level of exposure of fibronectin inside and on the surface of granulocytes and prognostic accuracy of the classifier from these parameters at the level of 100% using the method of support vector machine, SVM. High levels of diagnostic informativeness were recorded for the tests of all types of analyzed leukocytes with cell-associated fibronectin, and the classifiers based on the pair combinations of the tests with cell-associated fibronectin and fibronectin localized within the cells provide high diagnostic accuracy of the prognosis. Because the mentioned indicators are highly-sensitive tests, they can be proposed for early diagnostics and evaluation of the effectiveness of the conducted therapy of chronic diffuse liver diseases, which would allow reducing the use of paracentetic trepanobiopsy, a painful and risky procedure, which still remains the main type of diagnostic.

scholarly journals Aberrant UBR4 expressions in Hirschsprung disease patients

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Gunadi ◽  
Alvin Santoso Kalim ◽  
Estelita Liana ◽  
Aditya Rifqi Fauzi ◽  
Dian Nirmala Sirait ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Anorectal Malformation ◽  
E3 Ligase ◽  
Hirschsprung Disease ◽  
Control Group ◽  
Chain Reaction ◽  
Expression Change ◽  
Polymerase Chain ◽  
Hscr Patient

Abstract Background Recently, pathogenic alleles within ubiquitin N-recognin domain-containing E3 ligase 4 (UBR4) gene have been shown to be associated with Hirschsprung disease (HSCR). We determined the UBR4 expressions in Indonesian HSCR patients. Methods We analyzed the UBR4 expressions in the colons of HSCR patient and anorectal malformation (ARM) patient as control by real-time polymerase chain reaction (qPCR). Results Thirty-seven patients with non-syndromic HSCR and eighteen controls were involved in this study. qPCR revealed that the UBR4 expression was strongly decreased (0.77-fold) in the ganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.43 ± 0.36 vs. 2.05 ± 0.69; p = 0.009), whereas the UBR4 expression was also significantly reduced (0.79-fold) in the aganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.39 ± 0.46 vs. 2.05 ± 0.69; p = 0.044). However, the UBR4 expression change was not associated with gender (p = 0.35 and 0.80), nor with degree of aganglionosis both in ganglionic and aganglionic colons (p = 0.72 and 0.73), respectively. Conclusion Our study demonstrates that expression of UBR4 is decreased in both aganglionic and ganglionic colon of HSCR patients.

Mycoplasma pneumoniaeandChlamydophila pneumoniae: a comparative study in patients with nasal polyposis and healthy controls

2015 ◽  
Vol 129 (9) ◽  
pp. 865-869 ◽  
Author(s):  
D G Ioannidis ◽  
V A Lachanas ◽  
Z Florou ◽  
J G Bizakis ◽  
E Petinaki ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Inferior Turbinate ◽  
Sinus Surgery ◽  
Control Group ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain

AbstractIntroduction:The role played byMycoplasma pneumoniaeandChlamydophila pneumoniaein the pathogenesis of chronic rhinosinusitis with nasal polyps has been the object of ongoing debate. We used real-time polymerase chain reaction to investigate the prevalence of both microorganisms in the nasal tissue samples of patients and controls.Methods:We extracted DNA from nasal polyp samples obtained during functional endoscopic sinus surgery and the inferior turbinate samples of controls undergoing septoplasty. We used the highly sensitive real-time polymerase chain reaction to detect the presence ofM pneumoniaeandC pneumoniaeDNA.Results:Patients with chronic rhinosinusitis with nasal polyps consisted of 62 individuals (39 men; mean age 51 years); the control group consisted of 24 individuals (13 men; mean age 45 years). All samples from both groups were negative forM pneumoniaeandC pneumoniaeDNA.Conclusion:We have demonstrated that the likelihood ofM pneumoniaeandC pneumoniaeacting as an ongoing inflammatory stimulus in chronic rhinosinusitis with nasal polyps is slim.

Molecular Study of Cytomegalovirus Infection among Children with End Stage Renal Diseases Undergoing Dialysis, Pilot Study

2017 ◽  
Vol 2 (4) ◽  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Immunoglobulin M ◽  
Renal Diseases ◽  
Control Group ◽  
Chain Reaction ◽  
Specific Test ◽  
Polymerase Chain ◽  
End Stage

Cytomegalovirus is considered as an opportunistic infection affecting immunocompromized patients. Children with end stage renal diseases requiring dialysis is among affected population by this virus. The aim of the present study was to detect and compare the seroprevalence of CMV and CMV antigen pp65 with real time polymerase chain reaction (PCR) among children with end stage renal diseases undergoing dialysis. The study is a prospective case - control study. The forty one patients included in the studied are registered in the hospital for regular dialysis waiting for renal transplantation. The study included forty one healthy controls with same age and gender distribution. Blood samples were obtained from studied children and subjected for determination of specific immunoglobulin M and G for CMV (IgM-CMV, IgG-CMV) by Elecys system and CMV-DNA determination by real time polymerase chain reaction (PCR) and for PP65 antigenaemia test by light diagnostic CMVpp65. CMV-IgM was significantly detected frequently (P=0.0001) in 12.2% of the patients and in 2.4% of the control children. Moreover, IgG-CMV was significantly more frequently detected in patients (P=0.0001) than in control (90.2%&31.7% respectively). CMV-DNA was significantly (P=0.0001) detected in 12 patients (29.3%) compared to the control (2.4%), while CMVpp65 was detected among 4 children (9.8%) compared to one child in the control group. The comparison between IgM-CMV and real time PCR revealed that 30.7% of positive samples by PCR had positive IgMCMV, while IgG-CMV was associated with 84.6% of positive PCR. CMVpp65 correctly identified all negative samples compared to PCR, while the majority of negative PCR was also negative for IgM-CMV (98.6%). Moreover, all negative children for CMVpp65 was also negative by PCR (100%) For the validity of different CMV markers, IgG-CMV was the most sensitive test (84.7%), CMVpp65 was the most specific test 100%. From this study we concluded that CMV is a common viral infection among children with end stage renal diseases requiring dialysis. The diagnostic performance of real time PCR is the gold standard technique in diagnosis of this infection. CMVpp65 antigenemia is a specific accurate test for laboratory diagnosis however, it lacks sensitivity. Specific IgG for CMV is good screening diagnostic test.

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