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11 result(s) for "Krakowski, Leszek"
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BK Polyomavirus—Biology, Genomic Variation and Diagnosis
The BK polyomavirus (BKPyV), a representative of the family Polyomaviridae, is widespread in the human population. While the virus does not cause significant clinical symptoms in immunocompetent individuals, it is activated in cases of immune deficiency, both pharmacological and pathological. Infection with the BKPyV is of particular importance in recipients of kidney transplants or HSC transplantation, in which it can lead to the loss of the transplanted kidney or to haemorrhagic cystitis, respectively. Four main genotypes of the virus are distinguished on the basis of molecular differentiation. The most common genotype worldwide is genotype I, with a frequency of about 80%, followed by genotype IV (about 15%), while genotypes II and III are isolated only sporadically. The distribution of the molecular variants of the virus is associated with the region of origin. BKPyV subtype Ia is most common in Africa, Ib-1 in Southeast Asia, and Ib-2 in Europe, while Ic is the most common variant in Northeast Asia. The development of molecular methods has enabled significant improvement not only in BKPyV diagnostics, but in monitoring the effectiveness of treatment as well. Amplification of viral DNA from urine by PCR (Polymerase Chain Reaction) and qPCR Quantitative Polymerase Chain Reaction) is a non-invasive method that can be used to confirm the presence of the genetic material of the virus and to determine the viral load. Sequencing techniques together with bioinformatics tools and databases can be used to determine variants of the virus, analyse their circulation in populations, identify relationships between them, and investigate the directions of evolution of the virus.
BKPyV—Co-Architect of the Fate of a Renal Transplant During a One-Year Observation Period
To identify BKPyV, the VP1 protein sequence was analyzed and classified into genotypes in 246 RTRs before and after RTx from deceased donors during a one-year observation period. Quantitative assessment of BKPyV was conducted via qPCR. Prior to RTx, genotypes I and IV were identified in the urine (7.27 × 106; 1.20 × 105) and in serum (5.75 × 104; 1.12 × 104). After RTx, genotype I was predominant; identification of DNAuria-BKPyV (62.07%) and BKPyV-DNAemia (55.56%) peaked after three months, and the highest DNAuria-BKPyV titer was also observed after three months (6.48 × 109), whereas the BKPyV-DNAemia titer did not peak until after six months (2.21 × 107). The highest number of copies of genotype IV in the urine was observed after six months (9.54 × 109), while the highest titer in the serum was not observed until after 12 months (3.88 × 106). DNAuria-BKPyV precedes BKPyV-DNAemia, affects a larger group of patients, and has a greater and more easily detected viral load, which makes it not only an earlier marker, but the key predictive marker of greater clinical value than later detection of BKPyV-DNAemia alone. Early monitoring of DNAuria-BKPyV should be the basis of classical screening, and not merely an addition to it, and therapeutic interventions should be undertaken early to prevent nephropathy.
Molecular Epidemiology and Variation of the BK Polyomavirus in the Population of Central and Eastern Europe Based on the Example of Poland
The BK polyomavirus (BKPyV) is a widespread pathogen in humans. Polymorphism of the region encoding the VP1 protein of BKPyV provides the basis for classifying the virus into types and subtypes, whose frequency varies depending on geographic location. The aim of our study was to determine the frequency of BKPyV in the Polish population and to assess its variation by analysing polymorphism in the typing region. The study was conducted on 168 healthy, Polish volunteers, whose blood (plasma) and urine were sampled. The virus was detected using PCR, products, sequenced and subjected to bioinformatic analysis. In addition, viral load was assessed by qPCR. The presence of the genetic material of the BK virus was noted in 61/168 urine samples but in none of the plasma sample. Sequencing and phylogenetic analysis confirmed that the BKPyV isolates were of types I and IV, dominant in Europe (63.93% and 36.07%, respectively). All isolates from genotype I belonged to subtype Ib-2, showing polymorphism at position 1809 with a frequency of 61.54% (G1809A) and 38.46% (G1809C). To the best of our knowledge, this is the first study of this magnitude on the genetic variation of BKPyV among healthy volunteers in Poland.
Cytokine inflammatory response in dairy cows with mastitis caused by Streptococcus agalactiae
The aim of the study was evaluation of the concentrations of interleukin (IL)-1β, IL-8, IL-12β and tumour necrosis factor alpha (TNF-α) in the serum and milk of cows with mastitis caused by . A total of 60 milk samples from diseased cows and 30 milk samples from healthy cows were included in the study. Blood and milk samples were taken from Holstein-Friesian cows from three herds (two in tie-stall and one in a free-stall housing system) in Lublin Province in Poland. The concentrations of cytokines in blood serum and quarter milk samples were determined by ELISA. The levels of IL-1β, IL-8, IL-12β and TNF-α were significantly higher in the milk of cows suffering from mastitis caused by compared to the milk of healthy cows (263.03 55.36 pg/mL, 298.34 131.82 pg/mL, 604.10 139.17 pg/mL and 460.86 78.82 pg/mL, respectively). In the group of sick cows, cytokine levels were significantly higher in milk than in serum (263.03 55.25 pg/mL for IL-1β, 298.34 164.22 pg/mL for IL-8, 604.10 70.34 pg/mL for IL-12β and 460.86 104.78 pg/mL for TNF-α). The results confirm the involvement of the entire bovine immune system to protect against the bacteria first locally in the udder. The response of the mammary gland to infection caused by is rapid and already very strong at the beginning of the infection.
The influence of probiotic administration on the phagocytic and oxidative burst activity of neutrophils and monocytes in the peripheral blood of dairy cows during different lactation periods
The objective of this bovine peripheral blood study was a comparative assessment of the phagocytic activity of neutrophils and monocytes and of the intracellular killing capacity of neutrophils from cows given no probiotic and from cows which were administered a probiotic consisting of and . These activity types were compared during different lactation periods. A cohort of 20 pregnant dairy cows was divided into two groups of 10. The experimental group consisted of cows fed a ration supplemented with probiotics, and the control group consisted of cows fed an unsupplemented ration. Blood was drawn six times: 7 days before dry off, 14 days before parturition, and 7, 21, 60 and 90 days postpartum (DPP). The phagocytic activity of neutrophils and monocytes and the oxidative burst activity of neutrophils were determined by flow cytometry in the peripheral blood of all examined cows. Phagocytosis testing revealed increased percentages of phagocytic neutrophils and monocytes in the experimental group at 21, 60 and 90 DPP (P-value < 0.01). The mean fluorescence intensity (MFI) values for neutrophils and monocytes were higher on all days of the study (P-value < 0.01). In oxidative burst testing, the percentages of detected neutrophils and their MFI were increased in the experimental group on all days (P-value < 0.01). The use of probiotics supported dairy cows' immunity throughout the whole experiment. Probiotic supplementation may limit the occurrence of infectious diseases in these animals.
The influence of probiotic administration on selected leukocyte subpopulations and the serum amyloid A concentration in the peripheral blood of dairy cows during different lactation periods
The aim of the study was to compare selected leukocyte subpopulations and the serum amyloid A (SAA) concentration in the peripheral blood of cows at different stages of lactation. The blood of cows receiving a probiotic as a dietary supplement was compared with the blood of cows not receiving it. The research was conducted on 20 pregnant dairy cows randomly divided into two groups of 10 cows each. The experimental group consisted of cows given the probiotic as a feed supplement. The control group consisted of cows that were fed without supplementation. Blood was drawn six times for testing: 7 days before drying; 14 days before parturition; and 7, 21, 60 and 90 days postpartum. Leukocyte immunophenotyping was performed by flow cytometry. The blood of cows administered the probiotic revealed an increased percentage of forkhead box protein 3 (Foxp3) , T CD4 and B CD25 lymphocytes and β2 CD18 and αM CD11b integrins, and persistently low SAA levels at all time points. The activity of the immune system in cows receiving the probiotic was higher than in control cows. However, the stabilisation of the immune system of the supplemented cows may be indicated by the persistence of a low level of SAA throughout the experiment. Therefore, it can be assumed that the immune system of cows treated with the probiotic more easily adapts to changes in conditions in particular lactation periods and that these cows become more resistant to infectious diseases.
Assessment of selected immunological parameters in dairy cows with naturally occurring mycotoxicosis before and after the application of a mycotoxin deactivator
Mycotoxins in dairy cows can cause many non-specific symptoms often resulting from immune system overreaction. The study assessed the concentration of selected cytokines and acute phase proteins (APP) in cows with natural mycotoxicosis before and after using a mycotoxin neutraliser. The cytokines were tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 10 (IL-10), and the APP were serum amyloid A (SAA) and haptoglobin (Hp). The research was carried out on an experimental group (Exp) of 10 herdmate Holstein-Friesian cows with mycotoxicosis. The control group (Con) was 10 healthy cows of the same breed from a different herd. Cows in the Exp group were administered the mycotoxin deactivator Mycofix for three months. Blood was drawn from Exp cows once before administering Mycofix and a second time after three months of its use. Blood was also drawn from Con cows at the same times. Serum levels of TNF-α, IL-6, IL-10, SAA and Hp were assessed using ELISA. The concentrations of all cytokines and Hp in Exp cows were higher before treatment (P < 0.001) than those in Con cows. After three months of administering Mycofix, the concentrations of TNF-α and IL-6 were significantly lower than their pre-treatment levels (P < 0.001). The concentrations of IL-6, IL-10, and Hp were still significantly higher than those in the Con group (P < 0.001). In cows with mycotoxicosis, simultaneous stimulation of antagonistic processes was noted: a pro-inflammatory process in the upregulation of TNF-α and IL-6, and an anti-inflammatory one in the upregulation of IL-10. Despite the absorbent's use and the resolution of clinical symptoms in Exp cows, high levels of IL-10 and Hp and IL-6 were maintained. Assessment of the level of cytokines and APP appears to be a useful and precise tool for the evaluation and application of the appropriate dose of the mycotoxin absorbent or the evaluation of its effectiveness.
Changes in blood lymphocyte subpopulations and expression of MHC-II molecules in wild mares before and after parturition
Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents. The study was carried out on 24 Polish Konik breed mares which were divided into two equal groups. The first group (group I) included mares living in the reserve. The second group (group II) comprised mares maintained under conventional conditions in the stables. The blood samples were collected for the first time in the perinatal period, . 2 weeks before parturition (trial 0), then within the first 24 h after delivery, and then on 7 and 21 day after foaling. Flow cytometric analysis of lymphocyte expressing TCD4+, TCD8+, CD2+, and MHC class II antigens was performed. Before the delivery, in group I there was a significantly higher CD4:CD8 ratio compared to group II (P ≤0.05). Similarly, significantly increased CD4:CD8 ratio in group I was noted within 24 h after parturition (P ≤0.001) and it was also observed on 7 day (P ≤0.03) and 21 day after foaling (P ≤0.02). In the first 24 h after parturition, a significant decline of lymphocytes CD8+ (P ≤0.02) was noted. No significant differences in terms of lymphocytes CD2+ and CD3+ were observed. Expression of MHC-II molecules before and after the parturition was higher in group I compared to group II; however, the difference between the groups was not significant. The results obtained indicate that mares living in the reserve display higher activity of cell defence mechanisms.
Inflammatory cytokine and acute phase protein concentrations in the peripheral blood and uterine washings of cows with subclinical endometritis in the late postpartum period
The aim of the study was to evaluate the concentrations of proinflammatory cytokines: tumor necrosis factor (TNF-α) and interleukin-6 (IL-6), anti-inflammatory cytokine interleukin-10 (IL-10), and acute phase proteins (APPs)—haptoglobin (Hp) and serum amyloid A (SAA) in serum and uterine washings of cows with subclinical endometritis, and compare them to healthy animals. The study was performed on 24 cows on day 60 after delivery. The cows were divided into two groups based on the results of cytological tests: 12 cows with subclinical endometritis and 12 healthy cows. Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines in the study material were determined with ELISA: TNF-α, IL-6, IL-10 and APPs - Hp and SAA. The results show that the levels of TNF-α ( p  < 0.01), IL-6, IL-10 as well as SAA and Hp were significantly higher in the serum of cows with subclinical endometritis compared to the controls ( p  < 0.001). Uterine washings had significantly higher levels of IL-6, IL-10, and Hp in the experimental cows compared to the controls ( p  < 0.001). The demonstrated differences in the concentration of cytokines and APP between cows with subclinical endometritis and healthy cows, in both the serum and uterine washings, may suggest the usefulness of these parameters in the diagnosis of subclinical endometritis in cows in the late postpartum period.
Genotypes and Variants of BKPyV in Organ Donors after Brain Death
Kidney transplantation from a donor with latent BKPyV might be the cause of serious complications, such as BK virus-associated nephropathy. The aim of the study was to determine the prevalence of BKPyV infection in donors after brain death (DBDs), to analyse the molecular variation of BKPyV and to compare clinical and inflammation parameters of DBDs infected with various genotypes of BKPyV. BKPyV was investigated in blood and urine samples of 103 DBDs using PCR followed by sequencing and bioinformatic analysis, and the viral load was assessed by qPCR. Clinical parameters, including cellular markers of inflammation were assessed. The results confirm high prevalence of BKPyV (48%),and genotype IV (49%) over genotype I (43%) and the co-infection with genotypes I and IV in 8.2%. Viral load ranged from 102 to 107 copies/mL, with an average of 1.92 × 106 copies/mL. No specific markers for BKPyV infection were detected among the parameters tested. Infection with genotype I may be associated with the adverse impact on thekidney function, while infection with genotype IV was associated with the anemia Not only the viral load but also the genotype of BKPyV may have an impact on the course of infection.