Antioxidant capacity in renal transplant patients.

The aim of the study was to analyse the relation between total antioxidant capacity and immunosuppressive therapies, renal function and hematocrit in kidney transplant patients. The study included 46 adult patients during the maintenance period (>1 year) following renal transplantation, treated with different combinations of immunosuppressive agents--most commonly with cyclosporine (n = 23) or tacrolimus (n = 15). The total antioxidant capacity (TAOC) of plasma was measured using Trolox-equivalent antioxidant capacity (TEAC) assay. Patients treated with cyclosporine had significantly greater TAOC compared with those treated with tacrolimus (1.16 +/- 0.46 mmol/L vs. 0.80 +/- 0.37 mmol/L, p = 0.018, respectively). There was a significantly negative correlation between TAOC and plasma creatinine (rs = -0.551, p = 0.033) and a positive correlation between TAOC and creatinine clearance or hematocrit in patients treated with tacrolimus but not with cyclosporine (r = 0.525, p = 0.045 or rs = 0.629, p = 0.012, respectively). Immunosuppressive therapy with cyclosporine was associated with higher TAOC. Anemia can be an independent risk factor for an increase of oxidative stress. Although subject numbers werelimited, TAOC was positively associated with renal function in patients treated with tacrolimus.

The value of tumor marker and cytokine analysis for the assessment of regional lymph node status in cervical cancer patients.

The aim of the study was to evaluate the utility of the measurements of the circulating tumor markers, squamous cell carcinoma antigen (SCCA), CA125, carcinoembryonic antigen (CEA), cytokeratin fragment 19 (CYFRA 21.1), and the cytokines, interleukin-6 and vascular endothelial growth factor (VEGF), to estimate regional lymph node involvement in patients with cervical cancer. The study comprised 182 untreated patients with cervical cancer. The regional lymph node status was assessed either by the postsurgical histopathologic examination or by the computed tomography (CT). Concentrations of SCCA, CEA, and CA125 were determined using the Abbott Instruments system, of CYFRA 21.1 by the Roche kits, and of IL-6 and VEGF by the ELISA of R&D Systems (Minneapolis, MN). For the statistical analyses, Mann-Whitney U test and chi(2) test were applied. Serum levels of SCCA, CEA, CA125, CYFRA 21.1, IL-6, and VEGF were measured in patients with specified pelvic and para-aortic lymph node status. SCCA, CA125, and IL-6 levels were found to be significantly higher in patients with lymph node metastases than in those with no lymph node involvement. Also, the percentage of patients with simultaneously elevated concentrations of SCCA and CA125 or SCCA and IL-6 differed depending on the lymph node status and was significantly higher in the series of patients with lymph node metastases. Simultaneous assessment of serum levels of SCCA and CA125 or SCCA and IL-6 in patients with cervical cancer may be useful for the regional lymph node evaluation, especially in patients with advanced stages, when the lymph nodes are examined only by CT, with no histologic confirmation.

Cerebrospinal fluid and serum IL-8, CCL2, and ICAM-1 concentrations in astrocytic brain tumor patients.

BACKGROUND: The aim of the study was the evaluation of serum and CSF concentrations of CCL2, IL-8, and sICAM-1 in patients with astrocytic tumors as compared to a group of non-tumoral patients. METHODS: Chemokine concentrations were measured using the ELISA method. RESULTS: Regardless of the parameter tested and the patient group (brain tumor or non-tumoral patients), statistical differences (P < 0.05) were found between concentrations obtained in CSF compared to values obtained in serum for all proteins tested. CSF IL-8 concentrations were significantly elevated in CNS tumor patients as compared to non-tumoral individuals (P = 0.000); serum CCL2 and sICAM-1 concentrations were significantly decreased in CNS tumors in comparison with the comparative group (P = 0.002 and P = 0.026, respectively). Among proteins tested in the serum, a higher area under the ROC curve (AUC) revealed CCL2 compared to sICAM-1 in differentiating subjects with CNS brain tumors from non-tumoral subjects. AUC for CSF IL-8 was higher than for its index (CSF IL-8/serum IL-8). CONCLUSIONS: For individual biomarkers (IL-8 and CCL2, sICAM-1), measured in CNS brain tumor patients, the appropriate material, respectively CSF or serum, should be chosen and quantitatively tested. Increased cerebrospinal fluid IL-8 with decreased serum CCL2 create a pattern of biomarkers, which may be helpful in the management of CNS astrocytic brain tumors.

Inactivation of S-adenosylhomocysteine hydrolase during adenine arabinoside therapy.

To assess for possible inhibition of cellular transmethylation during adenine arabinoside (ara-A) therapy, S-adenosylhomocysteine hydrolase activity was analyzed in 10 patients with chronic hepatitis B virus infection. In six patients receiving ara-A, enzyme activity was suppressed to 0-2% of control erythrocyte enzyme activity. This decrease in enzyme activity was evident within 4 h of starting the drug infusion and continued for 7 d after cessation of therapy. S-adenosylhomocysteine hydrolase activity of peripheral mononuclear cells was also measured in two patients receiving ara-A. Suppression to as low as 3.5% of pretreatment levels was found; however, marked fluctuations with partial return of enzyme activity during therapy was also observed in mononuclear cells. Inhibition of an enzyme involved in transmethylation reactions was observed in patients during ara-A therapy. This could contribute to the side effects and antiviral properties of ara-A.

Peritoneal fluid cytokines and the differential diagnosis of benign and malignant ovarian tumors and residual/recurrent disease examination.

This study aimed to assess the potential value of peritoneal fluid cytokine examination for the differential diagnosis of ovarian tumors and for evaluating residual or recurrent disease after treatment. The cytokines that are commonly elevated in ovarian cancer, VEGF, IL-6, bFGF, IL-8 and M-CSF, and a reference ovarian tumor marker, CA 125, were measured in peritoneal fluids of 53 previously untreated patients with epithelial ovarian cancer, 18 ovarian cancer patients after surgical treatment and chemotherapy, and 17 patients with benign epithelial ovarian tumors. Non-parametric statistical analysis of data was performed. Ovarian cancer peritoneal fluids, as compared to peritoneal fluids of patients with benign ovarian tumors, contained significantly higher concentrations of IL-6, VEGF and CA 125, and significantly lower concentrations of bFGF and M-CSF, but only the levels of IL-6 and VEGF were significantly higher in peritoneal fluids of stage I and II ovarian cancer patients than of patients with benign ovarian conditions. IL-6 at the cutoff level of 400 pg/mL discriminated benign and malignant ovarian tumors with 92% sensitivity and 60% specificity, while VEGF at the cutoff of 400 pg/mL had 90% sensitivity and 80% specificity. At the cutoff level of 1200 pg/mL, IL-6 had 84% sensitivity and 87% specificity. A radical decrease in local cytokine and CA 125 levels in patients after treatment was independent of therapy outcome. IL-6 and VEGF measurements in peritoneal fluids might be useful for the differential diagnosis of malignant and benign ovarian conditions, but not for residual or recurrent disease examination.

Increased "pregnancy-specific" beta1-glycoprotein in certain nonseminomatous germ cell tumors.

Schwangerschafts (pregnancy) protein No. 1 (SP1), a recently identified beta1-glycoprotein that occurs during pregnancy, was assayed in the sera of 97 men with germ cell tumors of the testis. SP1 was elevated at 11-440 ng/ml in 3 of 6 men with choriocarcinomas, in 5 of 17 men with teratomas or "teratocarcinomas" (embryonal carcinomas and teratomas), and in 5 of 50 men with embryonal carcinomas; the highest value in 143 patients with nonmalignant diseases was 9.1 ng/ml. None of 24 sera from men with seminomas and none of 5 sera from men with orchitis had elevated SP1. In the 1 patient examined, testicular choriocarcinoma SP1 had immunochemical and gel chromatographic properties similar to those of highly purified SP1 of placental origin.

WW domains of Rsp5p define different functions: determination of roles in fluid phase and uracil permease endocytosis in Saccharomyces cerevisiae.

Rsp5p, ubiquitin-protein ligase, an enzyme of the ubiquitination pathway, contains three WW domains that mediate protein-protein interactions. To determine if these domains adapt Rsp5p to a subset of substrates involved in numerous cellular processes, we generated mutations in individual or combinations of the WW domains. The rsp5-w1, rsp5-w2, and rsp5-w3 mutant alleles complement RSP5 deletions at 30 degrees. Thus, individual WW domains are not essential. Each rsp5-w mutation caused temperature-sensitive growth. Among variants with mutations in multiple WW domains, only rsp5-w1w2 complemented the deletion. Thus, the WW3 domain is sufficient for Rsp5p essential functions. To determine whether rsp5-w mutations affect endocytosis, fluid phase and uracil permease (Fur4p) endocytosis was examined. The WW3 domain is important for both processes. WW2 appears not to be important for fluid phase endocytosis whereas it is important for Fur4p endocytosis. In contrast, the WW1 domain affects fluid phase endocytosis, but it does not appear to function in Fur4p endocytosis. Thus, various WW domains play different roles in the endocytosis of these two substrates. Rsp5p is located in the cytoplasm in a punctate pattern that does not change during the cell cycle. Altering WW domains does not change the location of Rsp5p.

The growth of mdp1/rsp5 mutants of Saccharomyces cerevisiae is affected by mutations in the ATP-binding domain of the plasma membrane H+ -ATPase.

Mutations in the PMA1 gene, encoding plasma membrane H+ -ATPase, were isolated that are able to suppress the temperature sensitivity (ts) phenotype of mdp1 mutations located in RSP5, the ubiquitin-protein ligase gene. The mdp1 mutants were previously found to change the mitochondrial/cytosolic distribution of Mod5p-I, the tRNA modifying enzyme, and to affect fluid phase endocytosis. The data presented reveal that mdp1 mutants are also pH sensitive, and hypersensitive to hygromycin B and paromomycin. The ts phenotype, hygromycin B and paromomycin sensitivity are suppressed by pmal-t, but the pH sensitivity, the effect of mdp1 on Mod5p-I cytoplasmic/mitochondrial localization and endocytosis are not. Characterization of pmal-t revealed the substitution of amino acid G(653)V in the ATP-binding domain of the H+ -ATPase. Our results indicate that Rsp5 ubiquitin-protein ligase may also influence, in addition to protein distribution, the functioning of plasma membrane H+ -ATPase and the response of cells to stress.

Simultaneous protective and damaging effects of cysteamine on intracellular DNA of leukocytes.

Incubation of human leukocytes with cysteamine can lead to the induction of DNA strand breaks. The induction of breaks is biphasic with increasing concentration of scavenger. The number of breaks increases in a dose-dependent manner to a maximum and then decreases at higher concentrations. Catalase has been shown to prevent the production of breaks, indicating an involvement of hydrogen peroxide. Cysteamine reacts with oxygen to generate hydrogen peroxide but at higher concentrations it also reacts with hydrogen peroxide. Thus, the biphasic effect of cysteamine on leukocyte DNA may be due to the sum of two separate reaction pathways. (i) Cysteamine reacts with oxygen to generate hydrogen peroxide which leads to DNA strand breakage. (ii) At higher concentrations, it eliminates hydrogen peroxide by reacting with it, thereby protecting the cellular DNA. Other antioxidant scavengers such as WR2721, acetylcysteine and ascorbate can also autooxidize to produce strand breaks. Thiourea and tetramethylurea do not. When tested for their ability to protect cells against DNA damage from added H2O2, the agent which most damaging by itself, cysteamine, was also the most protective.

Chemical modifications of alpha1,6-fucosyltransferase define amino acid residues of catalytic importance.

alpha1,6-Fucosyltransferase (alpha6FucT) of human platelets was subjected to the action of phenylglyoxal (PLG), pyridoxal-5'-phosphate/NaBH(4) (PLP), and diethyl pyrocarbonate (DEPC) the reagents that selectively modify the structure of amino acids arginine, lysine and histidine, respectively, as well as to N-ethylmaleimide (NEM), mersalyl, p-chloromercuribenzoate (pCMB), iodoacetate, iodoacetamide, and methyl iodide that react with sulfhydryl group of cysteine. In addition, we treated the enzyme with beta-mercaptoethanol, a reagent that disrupts disulfide bonds. All reagents except NEM significantly inactivated alpha6FucT. Protection against the action of PLG, PLP and sulfhydryl modifying reagents was offered by GDP-fucose, GDP, and the acceptor substrate, a transferrin-derived biantennary glycopeptide with terminal GlcNAc residues. Neither donor nor acceptor substrate offered, however, any protection against inactivation by DEPC or beta-mercaptoethanol. We conclude that arginine, cysteine and probably lysine residues are present in, or closely by, the donor and acceptor substrate binding domains of the enzyme, whereas histidine may be a part of its catalytic domain. However, the primary structure of alpha6FucT does not show cysteine residues in proximity to the postulated GDP-fucose-binding site and acceptor substrate binding site of the enzyme that contains two neighboring arginine residues and one lysine residue (Glycobiol. 10 (2000) 503). To rationalize our results we postulate that platelet alpha6FucT is folded through disulfide bonds that bring together donor/acceptor-binding- and cysteine- and lysine-rich, presumably acceptor substrate binding sites, thus creating a catalytic center of the enzyme.

Neutrophils promote the release of alpha-6-fucosyltransferase from blood platelets through the action of cathepsin G and elastase.

Human blood platelets release alpha-6-fucosyltransferase during coagulation of blood or after stimulation with thrombin or other agonists that cause platelet activation (Antoniewicz et al., FEBS Lett. 244 (1989) 388-390). However, in the absence of neutrophils the thrombin-stimulated platelets release only a small fraction of alpha-6-fucosyltransferase activity (Koscielak et al., Acta Biochim. Polon. 42 (1995) 35-40). We show that the effect of neutrophils is reproduced by cathepsin G or (less efficiently) by elastase, the two enzymes that are released by neutrophils during coagulation of blood. We have also localized alpha-6-fucosyltransferase to membrane and alpha-granule fractions of platelets that had been disrupted by nitrogen cavitation. It is concluded that thrombin-activated neutrophils release cathepsin G and elastase that promote degranulation of platelets and hence the secretion of alpha-6-fucosyltransferase.

Serum CA 15.3, CEA and ESR patterns in breast cancer.

Serum CA 15.3, CEA and ESR were longitudinally determined in 298 patients with breast cancer during postsurgical follow-up and/or therapy. Observation lasted until the death of the patient or at least for three years. With regards to longitudinal serum markers and ESR curves, four different patterns have been identified: pattern I: the markers and ESR stayed at normal levels; pattern II: the markers and ESR decreased from a peak level; pattern III: the markers and ESR fluctuated widely; pattern IV: the markers and ESR increased steadily. We have looked at over all survival (OS) and relapse-free survival ( RFS) versus longitudinal CA 15.3, CEA and ESR patterns. Univariate Cox regression analysis showed that OS and RFS were significantly associated with all four patterns.

Serum levels of carcinoembryonic antigen, gastrointestinal cancer-associated antigen and alphafetoprotein in staging and management of patients with advanced carcinoma of the stomach.

Serum levels of carcinoembryonic antigen (CEA), gastrointestinal cancer-associated antigen (GICA or CA 19-9), and alphafetoprotein (AFP) were concurrently determined in patients with carcinoma of the stomach: in 84 preoperatively, and in 67 serially postoperatively. Before surgery, serum CEA gave information about the tumor load analogous to serum GICA in 69% of the patients: true-positive in 25% and false-negative in 43%; less information in 18% and more in 14%. The sensitivity of the test tended to be better in the more advanced stages, and was higher for CEA with GICA than for CEA alone or GICA alone. During follow-up, serum CEA gave information about the presence or absence of active disease analogous to serum GICA in 78% of the patients: true-positive in 30%, true-negative in 36% and false-negative in 12%; less information in 9% and more in 13%. Neither test gave any false-positive indications. Sensitivity of the test rose from 67% for CEA alone and 60% for GICA alone to 81% for CEA with GICA. Serum AFP was elevated only preoperatively in 2% of patients. We conclude that joint application of CEA and GICA tests gave only slightly better preoperative sensitivity than CEA alone or GICA alone but proved fairly sensitive for postoperative follow-up of the patients. AFP was of little value for either purpose.

CRP, TNF-alpha, IL-1ra, IL-6, IL-8 and IL-10 in blood serum of colorectal cancer patients.

Blood serum cytokines: TNFalpha, IL-1ra, IL-6, IL-8, IL-10 as well as CRP were investigated in patients with colorectal cancer, prior treatment and 1, 10 and 42 days after surgery. There was an increase of the levels of CRP, IL-6 and IL-10 in most patients 24 hours after surgery. The levels of IL-1ra were elevated in patients in stage C and in several patients in stage B of the disease and there was a decrease of circulating TNFalpha in stage B patients. On day 10 and 42 after surgery, the levels of cytokines followed various patterns.

Mechanism of carnitine transport catalyzed by carnitine carrier from rat brain mitochondria.

The transport mechanism of reconstituted carnitine carrier purified from rat brain mitochondria was studied kinetically. Short and medium chain acyl carnitine derivatives had much higher affinity to the carnitine carrier in comparison with long chain acyl carnitine derivatives, therefore both homologous (carnitine/carnitine) and heterologous (carnitine/acetylcarnitine) antiports were analysed. A complete set of half-saturation constants was established for various substrate concentrations on both the external and the internal side of the membrane. Bisubstrate initial velocity analyses of the exchange reaction resulted in a kinetic pattern which is consistent with a sequential antiport mechanism. This type of mechanism implies formation of a ternary complex of the carrier with one internal and one external substrate molecule before the transport reaction occurs.

Usefulness of determination of CA-125 in monitoring patients with ovarian carcinoma.

CA-125 was determined in 298 patients with ovarian carcinoma both initially, and during and after treatment. The upper limit of normal for CA-125 in our laboratory is 65 units/ml. Postoperatively, serum CA-125 antigen showed three distinct patterns: 20% of patients in whom levels fluctuate within the normal range have active disease; 100% of patients with fluctuations above 65 U/ml are found to have active disease; 100% of patients in whom levels of CA-125 increase progressively from any nadir, have active disease. An elevated concentration of CA-125 antigen indicates presence of tumor and allows avoidance of 2nd-look surgery. Serum CA-125 antigen fluctuating within the normal range does not indicate eradication of the viable tumor cells.

Cytokines and endometriosis.

INTRODUCTION: Abnormal expression of cytokines is considered to be involved in the pathogenesis of endometriosis. MATERIAL AND METHODS: In 63 women with various stages of endometriosis preoperative levels of cytokines, including IL-6, IL-8, BVEGF, b-FGF, of TNF RI and TNF RII were analysed employing ELISA tests (R&D kits). RESULTS: In the sera of endometriosis patients significantly augmented levels of IL-6 and of b-FGF were detected as well as a trend for elevated IL-8 levels. CONCLUSION: Elevated serum levels of cytokines may show poor correspondence to the localized pathological process. Endometriosis would find a stricter reflection in cytokine levels in fluids or tissues of the pelvis minor.

[Frequency of human papillomavirus-herpes simplex-cytomegalovirus and Chlamydia trachomatis infection in genital tracts of sexual partners]. / Czestosc wystepowania infekcji wirusowych brodawczaka ludzkiego, opryszczki plciowej, cytomegalii oraz chlamydia trachomatis w narzadach plciowych partnerów seksualnych.

OBJECTIVES: Much attention is given nowadays to the role of Human Papillomavirus, Herpes simplex virus, Cytomegalovirus, and Chlamydia trachomatis--infections in cervical carcinogenesis. Cytomegalovirus, Herpes simplex and Chlamydia trachomatis are now thought to be teratogenic to humans. DESIGN: We investigated the prevalence of HPV, HSV, CMV and Chlamydia trachomatis in genital tracts of sexual partners. MATERIALS AND METHODS: 90 sexual partners were qualified for the research. Examination smears were taken with the dacron swab from the vaginal part of the uterine cervix, cervical canal, the lower vagina from women and from fossa navicularis penis in men. In the group of 67 men we have investigated semen as well. HPV, HSV, CMV and Chlamydia trachomatis were identified using PCR (Polymerse Chain Reaction)--method. RESULTS AND CONCLUSIONS: In 48% of investigated sexual partners we proved the presence of Human Papillomavirus, in 2.2% of women and 2.9% of men--Cytomegalovirus and in 11.1% of women and 14.9% of men--Chlamydia trachomatis. In the investigated biological material we did not find any HSV infection.

Assessment of the musculoskeletal load of the trapezius and deltoid muscles during hand activity.

The purpose of the study was to analyse the influence of the precision of a task on tension and fatigue of the trapezius and deltoid muscles. Ten young men took part in experiments. Different levels of force and different frequencies of pressing a button defined the precision of the task. Surface electromyography (EMG) was used. Muscle tension and fatigue were reflected by 2 parameters of the EMG signal: the Root Mean Square amplitude related to the maximum value and changes in the Median Power Frequency. The results showed that hand activities influence the descending part of the trapezius muscle and do not influence the deltoid muscle, and that the precision of work can influence the examined muscles of the arm and shoulder even during work in which only the hand is involved in a performed task.

Practical application of ergonomic settings of typical computerised workstations.

The goal of the study was to check, with regard to ergonomics, workstations equipped with visual display terminals in selected enterprises. Over 180 workstations were tested in 3 enterprises. Most workstations were equipped with computers. The ergonomics of both the parameters of the basic components of the workstation (i.e., a chair and a desk, and the position of the computer at the workstation and its screen with respect to windows) and lighting fittings were analysed. Typical mistakes in the layout of a workstation were chairs inappropriate for computer work, as well as broken chair adjustment mechanisms, which qualified chairs for repair or replacement. Wrong positioning of monitors on the desk and with regard to windows and lighting fittings was also noted.