Immunocytological urinalysis and monocyte chemotactic peptide-1 in renal transplant recipients with polyomavirus replication.

In some patients polyomavirus replication induces chronic tubulointerstitial inflammation in the transplanted kidney. The aim of this study was to investigate whether immunocytological urinalysis and monocyte chemoattractant protein-1 (MCP-1) assays could be used for an early diagnosis of nephropathy for patients with polyomavirus replication. We analyzed 1189 urine sediments from 174 renal allograft recipients who were transplanted between 2000 and 2005. Decoy cells were identified by an immunofluorescence method using specific antibodies (JC/BK monoclonal antibody). A similar method was used to detect CD3(+), CD14(+), and HLA-DR(+) cells with appropriate antibodies. The urinary excretion of MCP-1 was assayed by enzyme-linked immunosorbent assay. The results of urine sediment analysis and MCP-1 concentrations were compared with those of patients with stable graft function (control group n = 65). In 17 patients (10%) decoy cells were identified in urine. In 12 patients polyomavirus DNA was detected in plasma or urine by a polymerase chain reaction method. Polyomavirus nephropathy was diagnosed in eight patients by the presence of intranuclear viral inclusions or immunohistochemical staining with SV40 large T-antigen specimens from a renal biopsy, as well as by clinical and histopathological evidence (group I). Polyomavirus replication was diagnosed in four patients by urinary excretion of decoy cells and polyomavirus DNA detection (group III). In five patients only decoy cells were found. The patients of groups I and II showed an increased number of CD3, CD14, HLA-DR surface antigen-positive cells and greater excretion of MCP-1 compared with the control group (P < .02). The number of excreted cells was higher among patients with more severe infiltration. The results of patients from group III were similar to the control group. In conclusion, increased excretion of cells with CD3, CD14, and HLA-DR surface antigens and of MCP-1 were associated with intragraft tubulointerstitial inflammation in patients with polyomavirus nephropathy. Asymptomatic polyomavirus replication was associated with hidden tubulointerstitial inflammation. Monitoring cell excretion and chemokine content may be utilized for early detection of polyomavirus-induced nephropathy.

Limitations of in situ hybridization with total genomic DNA in routine screening for alien introgressions in wheat.

In situ hybridization with total genomic DNA (GISH) has become a powerful tool in characterization of alien introgressions in wheat. With recent simplification it can now be used in large scale screening for new chromosome constructs. Its level of resolution in routine applications was tested on sets of recombined wheat-rye chromosomes with genetically determined positions of the translocation breakpoints. The resolution level of GISH visualized by an enzymatic color reaction was much lower than that of GISH with fluorescent probes but both techniques failed to reveal the presence of some distally located breakpoints. The limits of resolution for the two methods were at least 9.8 and 3.5 cM of the relative genetic lengths of chromosome arms, respectively, in configurations with proximal rye and terminal wheat segments when rye DNA was used as a probe. When wheat DNA was used as a probe, a terminal wheat segment estimated to be ca. 1.6 cM in length could not be visualized. An example of induced recombination between a chromosome of Agropyron elongatum and wheat illustrates that these resolution limits of GISH may hamper isolation of critical translocation breakpoints in a chromosome engineering effort.

Innate antiviral immunity is not impaired in patients with chronic renal failure on hemodialysis.

UNLABELLED: Although it is well documented that chronic renal failure patients are susceptible to infectious diseases, the reason for this has not been clarified. The aim of the study was to assess the antiviral natural (innate) immunity of peripheral leukocytes in 37 hemodialysis patients and compare it with that of a of control group (70 blood donors). We investigated 16 patients with anti-hepatitis C virus (HCV) antibodies, anti-HCV(+) and 21 patients without anti-HCV antibodies, anti-HCV(-). METHODS: Innate immunity was measured using the method of direct infection of peripheral blood leukocytes with indicatory VS virus. The VS virus did not replicate in leukocytes with strong innate immunity, whereas by impaired immunity the virus multiplied to high titer. RESULTS: Patients on hemodialysis expressed the same levels of nonspecific antiviral immunity as the control group. We found complete, partial or deficient of innate immunity respectively in 33, 52.5, 14.5% of anti-HCV(-) patients, 43, 43 and 14% of anti-HCV(+) patients, and 44, 40 and 16% of controls. CONCLUSIONS: Innate antiviral immunity was not impaired (disturbed) in chronic HD patients. The categories of innate immunity disposition in dialysis patients and the healthy population did not differ.

The effect of a deficiency and a deletion on recombination in chromosome 1BL in wheat.

To test two models of chiasma allocation and the distribution of crossing-over in chromosomes, genetic mapping was performed in normal, deletion and deficiency chromosome arms 1BL of wheat, Triticum aestivum L. Shortening of the chromosome arm, either by a deletion of the proximal half of the arm or by a deficiency of the terminal quarter of the arm's length, significantly reduced the frequency of multiple crossovers but did not affect the distribution of the distal, presumably the first, crossover in the arm. In the deficiency chromosome, the recombination rate in the terminal segment was much higher than that in the same segment of the complete arm. This suggests that recombination frequency is not an inherent characteristic of a segment but depends on the segment's position on the centromere-telomere axis. These observations support the classical model of chiasma distribution along the chromosome based on the point of pairing initiation, chromosome length and the positive chiasma interference. The study also demonstrates that the distribution and frequency of recombination in a chromosome segment can be manipulated. Therefore, even the segments with very low recombination frequencies could be saturated with large numbers of crossover events to produce high-density genetic maps.

Cytokine-inducing activity of a proline-rich polypeptide complex (PRP) from ovine colostrum and its active nonapeptide fragment analogs.

A complex of proline-rich polypeptides (PRP) was isolated from ovine colostrum in our laboratory and was shown to possess immunomodulatory properties and psychotropic activity, including beneficial effects in the treatment of Alzheimer's disease. A nonapeptide fragment (NP): Val-Glu-Ser-Tyr-Val-Pro-Leu-Phe-Pro, isolated from the chymotryptic digestion products of PRP, and its C-terminal fragment, a hexapeptide (HP): Tyr-Val-Pro-Leu-Phe-Pro also exhibited immunoregulatory activity. Although NP and HP expressed activity similar to that of PRP in studies on humoral and cellular immune responses, in other immune processes, e.g. induction of cytokines, they showed markedly lower activity than PRP. In the search for more active peptides, in the present study, we compared the cytokine-inducing ability of PRP, NP, HP, and linear oligomers of NP or HP. For this purpose, the induction of IFN, TNF-alpha, IL-6, and IL-10 in human whole blood cell cultures was measured. NP, HP, and their oligomers showed differential effects in the induction of cytokines, generally lower than that of PRP. Only the PRP complex showed a bell-shaped dose-response dependence suggesting regulatory properties. There were no distinct differences between monomeric forms of NP (NP1) or HP (HP1) and their oligomers in the induction of IFN and TNF-alpha (Th1 cytokines) but such differences were found in the induction of IL-6 and IL-10 (Th2 cytokines). Dimer (NP2) was less active than the monomeric NP1 nonapeptide in the induction of IL-6 and IL-10. On the other hand, oligomers: HP3 and HP4, showed a significantly higher ability to induce Th2 cytokines compared to HP1, HP2 or NP peptides. This was especially evident in the case of IL-10 induction, where the activity of HP4 surpassed the activity of PRP and approached the activity of LPS-PHA. The results obtained showed that some of the peptides studied, when used at higher concentrations (100 microg/ml) may replace the PRP complex as cytokine inducers. Our data also suggest the possibility of using certain oligomers for selective induction of particular cytokines.

Chlorinated phenols as metabolites of lindane. Evaluation of the degree of conjugation in rat urine.

After lindane was administered to rats in a single oral dose, the time profile of the degree of conjugation of the main phenolic metabolites was evaluated. In all urine samples 2,3-dichlorophenol, 2,4,5- and 2,4,6-trichlorophenol, and 2,3,5,6-tetrachlorophenol were constantly present. 2,3,5,6-Tetrachlorophenol and then 2,4,6-trichlorophenol were the prevalent metabolites in all cases. The degree of conjugation did not correlate with the dissociation constant of individual chlorinated phenols. Phenol conjugation declined with time after administration.

[A sensitive method for detection of sympathomimetic amines in pentafluorobenzamide form in biological samples using gas chromatography]. / Citlivý prukaz sympatomimetických aminu v biologických vzorcích ve forme pentafluorobenzamidu pomocí plynové chromatografie.

A toxicological method of sensitive and specific confirmation of methamphetamine and other primary and secondary amines in biological samples after extractive perfluorobenzoylation is described. The method is based on the principle of gas chromatography with parallel specific nitrogen detection and electron capture detection. The other alternative is the gas chromatography combined with mass spectrometry in chemical ionization mode. The method described allows the detection of methamphetamine and amphetamine in urine in concentrations below 10 ng/ml.

Malignant melanoma in a patient with polyomavirus-BK-associated nephropathy.

A possible relationship between malignant melanoma and polyomavirus-associated nephropathy in a renal transplant patient is presented. It is possible that BK polyomavirus (BKV) nephropathy and malignant melanoma developed independently. The coincidence may be related to over-immunosuppression. The presented case suggests careful oncological surveillance in patients with BKV nephropathy.

[Selenoproteins--atypical function of the UGA codon]. / Selenoproteiny--nietypowa funkcja kodonu UGA.

Selenium is an essential trace element for animals and humans. It's a structural component of the active center of several important mammalian enzymes, like gluthatione peroxidase (GSH-Px), iodothyronine deiodinase, and others selenoproteins. Paper describes selenium functions in physiological states, and mechanism of incorporation of selenocysteine into selenoproteins. This incorporation requires the alternative decoding of UGA, which typically serves as a stop codon. The most important element is the secondary structure of mRNA named SECIS (selenocysteine inserstion sequence).

[Computerization of the orthopedic-traumatology department]. / Program komputerowy w pracy oddzialu urazowo-ortopedycznego.

The authors the scheme of the computerization of their orthopedic-traumatologic department. An everyday work has been markedly advanced.

Genetic and physical mapping of homoeologous recombination points involving wheat chromosome 2B and rye chromosome 2R.

Wide hybrids have been used in generating genetic maps of many plant species. In this study, genetic and physical mapping was performed on ph1b-induced recombinants of rye chromosome 2R in wheat (Triticum aestivum L.). All recombinants were single breakpoint translocations. Recombination 2RS-2BS was absent from the terminal and the pericentric regions and was distributed randomly along an intercalary segment covering approximately 65% of the arm's length. Such a distribution probably resulted from structural differences at the telomeres of 2RS and wheat 2BS arm that disrupted telomeric initiation of pairing. Recombination 2RL-2BL was confined to the terminal 25% of the arm's length. A genetic map of homoeologous recombination 2R-2B was generated using relative recombination frequencies and aligned with maps of chromosomes 2B and 2R based on homologous recombination. The alignment of the short arms showed a shift of homoeologous recombination toward the centromere. On the long arms, the distribution of homoeologous recombination was the same as that of homologous recombination in the distal halves of the maps, but the absence of multiple crossovers in homoeologous recombination eliminated the proximal half of the map. The results confirm that homoeologous recombination in wheat is based on single exchanges per arm, indicate that the distribution of these single homoeologous exchanges is similar to the distribution of the first (distal) crossovers in homologues, and suggest that successive crossovers in an arm generate specific portions of genetic maps. A difference in the distribution of recombination between the short and long arms indicates that the distal crossover localization in wheat is not dictated by a restricted distribution of DNA sequences capable of recombination but by the pattern of pairing initiation, and that can be affected by structural differences. Restriction of homoeologous recombination to single crossovers in the distal part of the genetic map complicates chromosome engineering efforts targeting genes in the proximal map regions.