immunoenzymatic method
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Biomolecules ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1816
Author(s):  
Beata Szymańska ◽  
Michał Matuszewski ◽  
Janusz Dembowski ◽  
Agnieszka Piwowar

Background: Uroplakins (UPs) are glycoproteins that play a specific role in the structure and function of the urothelium. Disorders which affect the normal expression of UPs are associated with the pathogenesis of infections and neoplasms of the urinary tract, primary vesicoureteral reflux, hydronephrosis and renal dysfunction. The appearance of uroplakins in the urine and/or plasma may be of potential importance in the detection of urinary tract dysfunction. The aim of the present study was to investigate uroplakin IIIa (UPIIIa) and uroplakin II (UPII) expression in patients with selected urological diseases. Methods: Plasma and urine from patients with benign prostatic hyperplasia (BPH), urethral stricture (US), urinary tract infection (UTI) and urolithiasis were compared to healthy people without urological disorders. UPs concentrations were measured by the immunoenzymatic method. Results: In patients with BPH and UTI, concentrations of UPIIIa in urine and plasma, as well as UPII in urine, were statistically significantly higher than in the control groups. In the US group, only the plasma UPIIIa concentration differed significantly from the control. Conclusion: The conducted research shows that benign urological diseases may affect the state of the urothelium, as manifested by increased concentrations of both UPs in patients’ urine and plasma, especially in BPH and UTI.


Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3100
Author(s):  
Jari Zambarbieri ◽  
Pierangelo Moretti ◽  
Alessia Giordano ◽  
Paola Scarpa

The determination of parathyroid hormone (PTH) in cats could be of clinical utility in many metabolic disorders, such as renal diseases, hypercalcemia, or nutritional imbalances. However, the available methods for the measurement of feline PTH are limited, not widely available, and need radioimmunoassays. The aim of this study was to perform the analytical validation of a new immunoenzymatic method for the measurement of feline PTH. Thirty-eight cats affected with chronic kidney disease (CKD) were included. PTH was measured using a two-site immunoenzymatic method validated in humans and dogs (ST AIA-PACK® Intact PTH, Tosoh Bioscience, Tessenderlo, Belgium). The analytical validation provided the evaluation of precision (intra-assay and inter-assay), accuracy (linearity under dilution (LUD) and spike recovery test (SRT)), and the storage stability of serum samples at 20 °C, 4 °C, and −20 °C. The method showed good precision (intra-assay CVs (coefficient of variations) 3.19–9.61%; inter-assay CVs 9.26–15.28%). In both the intra- and inter-assays, the highest imprecision was found with the low concentration pool (9.61% and 15.28%) and accuracy (LUD and SRT r2 = 0.99, p < 0.001), while the stability was optimal up until 7 days at −20 °C (−7.7%). The method was successfully validated in cats, allowing its future use in diagnostic procedures.


Biomolecules ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1422
Author(s):  
Michał Kukla ◽  
Tomasz Menżyk ◽  
Marcin Dembiński ◽  
Marek Winiarski ◽  
Aleksander Garlicki ◽  
...  

Analysis of liver biopsy specimens showed that SARS-CoV-2 might have led to liver damage. This study aimed to evaluate the role of selected hepatokines and myokines in the development and progression of COVID-19. Seventy patients with laboratory-confirmed COVID-19 and 20 healthy volunteers were enrolled in the study. Irisin, pentraxin 3, fetuin-A, and FGF-21 serum concentrations and biochemical parameters were assessed using an immunoenzymatic method with commercially available enzyme immunoassay (EIA) or enzyme-linked immunosorbent assay (ELISA) kits. Serum fetuin-A concentrations were significantly decreased in COVID-19 patients compared to healthy volunteers. The serum concentration of FGF-21 was significantly increased in obese COVID-19 patients compared to overweight ones. Moreover, the FGF-21 level was higher in COVID-19 patients diagnosed with metabolic syndrome than in patients without metabolic syndrome. PTX3 concentration was higher in COVID-19 patients with higher HOMA-IR values than those with lower HOMA-IR values. COVID-19 patients with HOMA-IR ≤3 and >3 had significantly lower fetuin-A levels than the control group. Irisin concentration was significantly decreased in the HOMA-IR ≤ 3 COVID-19 subgroup when comparing with the control group. Lower levels of fetuin-A observed in COVID-19 patients despite higher HOMA-IR, CRP, and ferritin levels, pneumonia, patients requiring ICU care suggests that fetuin-A deficiency predisposes to more severe COVID-19 course. Upregulated pentraxin 3 may be used as a potential predictor of COVID-19 severity.


2021 ◽  
Author(s):  
Beata Szymańska ◽  
Michał Matuszewski ◽  
Janusz Dembowski ◽  
Agnieszka Piwowar

Abstract BackgroundUroplakins (UPs) are glycoproteins that play a specific role in the structure and function of the urothelium. Disorders of normal expression of uroplakins are associated with the pathogenesis of infections and neoplasms of the urinary tract, primary vesicoureteral reflux, hydronephrosis and renal dysfunction. The appearance of uroplakins in the urine and/or plasma may be of potential importance in the detection of urinary tract dysfunction. The aim of the study was to investigate uroplakin IIIa (UPIIIa) and uroplakin II (UPII) expression in patients with selected urological diseases. Plasma and urine from patients with benign prostatic hyperplasia (BPH), urethral stricture (US), urinary tract infection (UTI) and urolithiasis, were compared to healthy people without urological disorders. MethodsA total 152 of human urine and plasma samples from normal and patients with selected benign urological diseases were analyzed. UPs concentration was measured by immunoenzymatic method. All calculations were done using the STATISTICA 13.3 (TIBCO software Inc.).ResultsIn patients with BPH and UTI, UPIIIa in urine and plasma also UPII in urine concentrations were statistically significantly higher than in the control groups. In the US group, only the plasma UPIIIa concentration differed significantly from the control. There were no significant differences between the concentrations of UPs compared to the controls in both the urine and plasma of patients with urolithiasis. ConclusionThe conducted research shows that benign urological diseases may affect the state of the urothelium, as manifested by an increased concentration of both UPs in patients’ urine and plasma, especially in BPH and UTI.


2021 ◽  
Vol 74 (10) ◽  
pp. 2384-2391
Author(s):  
Oksana Pavliuk ◽  
Sergii Shevchuk

The aim: To evaluate osteocalcin and PINP levels in men suffering from AS and to compare them with structural and functional state of bone tissue and clinical course of illness. Materials and methods: The study included 82 patients suffering from AS with an average age of 40,9±0,9 years. Osteocalcin level was determined in 82 patients, and PINP level was determined in 79 patients. Control group included 22 apparently healthy persons. Disease activity was assessed through CRP level, ASDAS and BASDAI scores, while functional ability was assessed through the BASFI score. Osteocalcin and PINP levels were determined by immunoenzymatic method for the purpose of evaluating the metabolic state of bone tissue. Results: Average osteocalcin and PINP levels were not significantly different in patients suffering from AS and patients in the control group and did not show any significant correlation with ASDAS, BASDAI, BASFI and CRP scores. In patients with spinal ankylosis, average osteocalcin values (14,3 ng\ml) and PINP (747,2 pg\ml) were higher compared to patients with single syndesmophytes (11,0 ng\ml; 711,8 pg\ml) and patients without syndesmophytes (10,4 ng\ml; 537,7 pg\ml respectively). Conclusions: Osteocalcin and PINP levels are not related to age, disease duration, BMI, glucocorticoids load and inflammatory process activity, however, they are closely related to the presence of bone growths.


Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2411
Author(s):  
Jari Zambarbieri ◽  
Filippo Tagliasacchi ◽  
Pierangelo Moretti ◽  
Alessia Giordano ◽  
Paola Scarpa

Renal hyperparathyroidism (RHPT) is one of the main complications in dogs affected with Chronic Kidney Disease (CKD). The measurement of serum parathyroid hormone (PTH) could be of clinical utility for the disease’s treatment and follow-up; however, PTH is not routinely determined due to limited available methods, often not fully validated in dogs. The aims of this study were the analytical validation of an immunoenzymatic method for the measurement of PTH in canine serum and the analysis of preliminary association of the obtained results with renal function. Twenty-six samples obtained from dogs healthy or affected with CKD were analysed. PTH was measured using a two-site immunoenzymometric human assay (ST AIA-PACK® Intact PTH, Tosoh Bioscience). The analytical validation protocol evaluated the assay precision and accuracy. Also, the PTH’s storage stability at 20 °C, 4 °C and −20 °C was assessed. Clinical validation was performed by comparing PTH values with creatinine, phosphorus and International Renal Interest Society (IRIS) stage. The method showed optimal precision and accuracy, whereas stability was adequate up to 4 h at 20 °C, 24 h at 4 °C and 6 months at −20 °C. PTH was positively associated with creatinine, phosphorus and IRIS stage. The investigated method was thus successfully validated in dogs, allowing its use for clinical purpose.


2018 ◽  
pp. 26-29
Author(s):  
S.O. Levenets ◽  
◽  
N.O. Udovikova ◽  
S.V. Novokhatskaya ◽  
D.A. Kashkalda ◽  
...  

The objective: to allocate the clinico-anamnestic and hormonal and metabolic indicators influencing the remote results of treatment Spironolactonum. Materials and methods. 78 girls at the age of 13–17.5 years with a hypomenstrual syndrome are surveyed (НMS: an oligomenorrhea – OHM, a secondary amenorrhea – VA) in 2 years and more after an initiation of treatment Spironolactonum (3 ten-day courses). The menstrual age (MA), the perinatal anamnesis, НMS duration prior to treatment, the index of body weight (IBW), uterus volume according to ultrasonic examination (US) of organs of a small pelvis, degree of expression of a hirsutism, contents in a cow of gonadotrophic, sex hormones, рrolactinum (PRL), insulin were studied by an immunoenzymatic method. The prognostic coefficient (PC) and its informational content were defined (I). The criterion of efficiency of treatment is an independent regular menstrual cycle. Results. The remote efficiency of treatment at OHM reached 62.9%, at VA – only 20.1% (Р < 0.001). The most prognostically significant relatively positive takes of treatment is the MV up to 2 years (PC = +5.13; I = 0.542), rather negative results – VA (PC = - 3.54; I = 0.545), a hirsutism of the II–III Art. on Ferrimana-Gаlvey’s scale, the increased LG level (PC = - 3.52; I = 0.59) and the lowered contents in blood estradiolum (PC = - 3.68; I = 0.36). Conclusions. The separated efficiency of treatment Spironolactonum within three months is 3 times better at OHM, than at VA. Girls with НMS demand inspection and treatment in the first 1–1.5 years after a menarche. Special attention should be paid to girls with the expressed hirsutism, to at what disorders of menstrual function remain within 2 years and more, with the LG high initial level and the lowered content in blood estradiolum. Key words: teenage girls, hypomanstrual syndrome, spironolactone, long-term effectiveness, гирсутизм, estradiol.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
B. Szymańska ◽  
M. Matuszewski ◽  
J. Dembowski ◽  
R. Zdrojowy ◽  
A. Długosz

Background. Uroplakins are glycoproteins investigated as potential markers of urothelial carcinoma. However, their role in chemical carcinogenesis is uncertain. In this study the diagnostic value of plasma and urine uroplakin IIIa (UPIIIa) levels in bladder cancer (BC) was investigated, particularly in the aspect of environmental exposure to chemical carcinogens, measured by DNA damage and detoxification ability in the BC smoking group. The correlation between uroplakin, 8-OHdG, and GSTπ was investigated. Material and Methods. This study included 61 BC patients and 33 healthy controls. UPIIIa, 8-OHdG, and GSTπ levels were estimated by the immunoenzymatic method (ELISA). Results. UPIIIa levels were elevated in BC patients in plasma (p≤0.001) and in urine (p≤0.001), as were 8-OHdG and GSTπ levels in urine. Moreover, the 8-OHdG level was higher in invasive or high grade tumors. A positive correlation between UPIIIa/GSTπ and 8-OHdG/GSTπ was observed, but no UPIIIa/8-OHdG correlation was noted. Conclusion. The study showed the diagnostic value of urine and plasma UPIIIa in BC (good sensitivity, specificity, and predictive value). The lack of UPIIIa correlation with 8-OHdG and smoking suggests that UPIIIa does not reflect the environmental exposure. The increased levels of 8-OHdG and GSTπ in the invasive tumor stage indicate their value in BC monitoring.


2014 ◽  
Vol 5 (1) ◽  
pp. ar.2014.5.0076 ◽  
Author(s):  
Giorgio Ciprandi ◽  
Mara DeAmici

Allergic rhinitis (AR) is characterized by Th2 polarized immune response. Soluble HLA (sHLA) molecules play an immunomodulatory activity. Two different studies evidenced that both patients with seasonal AR (SAR) and patients with perennial AR (PAR) had higher sHLA-G levels than normal controls. The aim of this study was to compare sHLA-G serum levels in SAR and PAR patients, also considering allergen-specific IgE. One hundred sixty-eight AR patients were enrolled, 94 with SAR and 74 with PAR. A group of 116 healthy subjects was considered as control. sHLA-G and allergen-specific IgE serum levels were determined by immunoenzymatic method. SAR patients had significantly higher levels of sHLA-G than PAR patients (p = 0.0194). sHLA-G was moderately related to allergen-specific IgE both in SAR (r = 0.497) and in PAR patients (r = 0.584). The present study provides evidence that sHLA-G serum levels depend on the type of allergy and are related to allergen-specific IgE serum levels. These findings may suggest that sHLA-G could be a biomarker of allergic reaction.


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