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Biomedical subjects

M Kurpisz

Publications and source records attributed to M Kurpisz.

At least 19 recordsLinked to original sources
PubMed1997

Analysis of HLA class Ib gene expression in male gametogenic cells.

We have investigated mRNA expression for nonclassical MHC class I genes (HLA-E,-F,-G) in human gametogenic cells. Testicular tissue was treated by collagenase and the resulting cell suspension was further purified by fractionation on Percoll gradients in a two-step procedure. Three gametogenic cell fractions were analyzed: purified heterogenous suspension of gametogenic cells, fraction of round spermatids and fraction of elongated spermatids. Total RNA isolated from each cell population was subjected to both reverse transcriptase/polymerase chain reaction and Northern blot analysis using oligonucleotides specific for HLA-E, -F and -G. Both method gave similar results. We have found a considerable level of HLA-E mRNA, very low amounts of reamplified cDNA for HLA-F and both a complete lack of mRNA and reamplified cDNA for the HLA-G gene in the analyzed gametogenic cell fractions. Additionally, we have localized HLA-E molecules on the cells of the adluminal compartment within seminiferous tubules using immunostaining with monoclonal antibodies specific for HLA-E heavy chain followed by confocal microscopy analysis. The unique expression pattern of HLA class I antigens in the male gonad could play an important role in an efficient protection against an autoimmunological attack toward germ cells.

Blotting, Northern↗
PubMed1997

Protection from acetaminophen-induced liver damage by the synergistic action of low doses of the poly(ADP-ribose) polymerase-inhibitor nicotinamide and the antioxidant N-acetylcysteine or the amino acid L-methionine.

1. An array of therapeutically used analgetic and antirheumatic drugs cause severe liver damage. The present study investigates the hepatoprotective effects of inhibitors of NAD-dependent adenoribosylation reactions and of antioxidants in analgesic-induced hepatic injury. 2. Male NMRI mice were treated PO with 500 mg/kg of acetaminophen, and the activities of both glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) were determined in serum. 3. The acetaminophen-induced release of both GOT and GPT from injured liver cells could be inhibited in a dose-dependent manner, when mice were injected additionally either with increasing amounts (from (25 mg/kg to 100 mg/kg i.p.) of the PARP-inhibitor nicotinamide, with increasing amounts (from 25 mg/kg to 100 mg/kg i.p.) of the antioxidant N-acetylcysteine, or with increasing amounts (from 50 mg/kg to 300 mg/kg i.p.) of the amino acid L-methionine. 4. A combination of both nicotinamide and N-acetylcysteine (at the low dose of 12.5 mg/kg i.p. each) results in a complete protection from acetaminophen-induced release of GOT and GPT from injured liver cells. 5. A combination of both L-methionine and N-acetylcysteine or nicotinamide (at the low dose of 12.5 mg/kg IP each) resulted also in complete protection from acetaminophen-induced release of GOT and GPT.

Acetaminophen↗
PubMed1997

Chromosomal anomalies as a predisposing factor for male infertility.

A "genetic factor" can be the reason for about 20% of male infertility cases. This could be due to chromosomal aberrations in sperm with immediate effect on conception or may result in recurrent spontaneous abortions of paternal origin. Techniques for the analysis of sperm chromosomal aberrations are still far from ideal. Sperm penetration to zona-free hamster oocytes and random in situ hybridization with sperm DNA using a range of fluorescent probes--are currently under investigation. Sperm chromosome analysis was performed in three distinct subgroups, (a) healthy, fertile individuals, (b) healthy but infertile subjects, and (c) carriers of chromosomal aberrations in lymphocytes. Sperm chromosomal aberrations occur in normal and fertile population in range from 1.9% to 15.8%. The aberrations are concerned both with number and structure of chromosomes. In infertile individuals, the main chromosome aberrations observed were translocations and pericentric inversions. Several hypotheses are also described indicating possible reasons for subfertility due to chromosomal anomalies. There are few gene families that can be involved in the regulation of spermatogenesis, mainly located on the Y-chromosome. Identification of defective gene(s) may lead to novel therapeutic strategies for infertility treatment.

Animals↗
PubMed1996

Influence of diet free of NAD-precursors on acetaminophen hepatotoxicity in mice.

Recently, we demonstrated the hepatoprotective effects of nicotinic acid amide, a selective inhibitor of poly(ADP-ribose) polymerase (PARP; EC 2.4.2.30) on mice suffering from acetaminophen (AAP)-hepatitis, suggesting that the AAP-induced liver injury involves a step which depends on adenoribosylation. The present study investigates the effects of a diet free of precursors of NAD, the substrate on which PARP acts, in female NMRI mice with AAP hepatitis and evaluates the influence of simultaneous ethanol consumption in these animals. Liver injuries were quantified as serum activities of glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT). While AAP caused a 117-fold elevation of serum transaminase activities in mice kept on a standard laboratory diet, which was significantly exacerbated by ethanol and inhibited by nicotinic acid amide (NAA), adverse effects were noted in animals fed a diet free of precursors of NAD. In these animals, only minor increases of serum transaminase activities were measured in the presence of AAP, and unlike the exacerbation caused by ethanol in mice on a standard diet, the liver damage was inhibited by 50% by ethanol. A further 64% reduction of hepatitis was observed, when NAA was given to ethanol/AAP-mice. Our results provide evidence that the AAP-induced hepatitis and its exacerbation by ethanol can either be reduced by end-product inhibition of PARP by NAA or by dietary depletion of the enzyme's substrate NAD. We see the main application of NAA as for the combinational use in pharmaceutical preparations of acetaminophen in order to avoid hepatic damage in patients treated with this widely used analgesic.

Acetaminophen↗
PubMed1996

Suppression of inflammatory arthritis by simultaneous inhibition of nitric oxide synthase and NADPH oxidase.

TH1-type proinflammatory cytokines induce the expression of phagocytic nitric oxide synthase (NOS) and prime the membrane-bound NADPH oxidase of neutrophils and monocytes of mice so as to attain an activated state, which upon a second stimulus releases up to 6-fold increased levels of reactive oxygen species (ROS) than do unprimed phagocytes. Enhanced levels of ROS and NO deregulate inflammatory signal transduction pathways, which play a crucial role in the pathogenesis of arthritis. The antiarthritic reactivity of diphenylene iodoniumchloride (DPI), an irreversible inhibitor of NADPH oxidase and NOS, was tested in male DBA/1xB10A(4R) hybrid mice suffering from potassium peroxochromate-induced arthritis. Daily doses of 2.8 mu mol/kg of DPI sufficed to inhibit the arthritis by 50%. A complete inhibition was obtained with 10 mu mol/kg of DPI. The reduction of overt arthritic symptoms correlated well with both the reduced levels of ROS and NO in plasma of DPI-treated mice. Our data support the hypothesis that oxidative stress and nitric oxides play a pivotal role in the pathology of arthritis, which can be therapeutically targetted by NADPH oxidase- and NO synthase-inhibitors.

Animals↗
PubMed1996

The easy formation of antisperm antibodies in prepubertal boys and the difficult humoral response in severe-combined immunodeficiency mice.

OBJECTIVE: To investigate and compare mechanisms controlling the development of antisperm antibodies in situations with different status of active immunosuppression. DESIGN: Antibody activity against human sperm was analyzed by immunosorbent assays (ELISA) in sera samples of prepubertal boys and in circulation of sensitized immunocompromised mice. SETTING: Procedures were performed in a university laboratory. PATIENT(S): Prepubertal healthy boys and patients with testicular failures. INTERVENTION(S): Blood was collected from healthy individuals and from patients with diagnosed testicular pathology; antisperm antibodies were rechecked after surgical intervention (orchidopexia or removal of the pathological gonad). MAIN OUTCOME MEASURE(S): Antisperm antibodies in quantitative ELISA. RESULT(S): Significant antibody activity to finally differentiated human sperm was detected in sera samples from prepubertal boys with testicular failures (cryptorchid or mobile testis), especially in individuals with both pathological gonads. "Naive" human lymphocytes deposited to peritoneal cavity of immunocompromised mice did not respond to in situ challenge with sperm antigens. CONCLUSION(S): High antisperm antibody levels in prepubertal boys may suggest low immunosuppressive activity at this age. This situation may influence future fertility status of these individuals. Immunocompromised mice effectively prevented humoral response to sperm antigens suggesting different mechanism than T suppressor cell activity.

Adolescent↗
PubMed1996

Seminal plasma can be a predictive factor for male infertility.

Seminal plasma from ejaculates of 10 healthy, fertile volunteers and 63 infertile males was analysed for superoxide dismutase (SOD) and xanthine oxidase (XO) activities using a chemiluminometer. There was not statistically significant difference in the activity of either enzyme between control and infertile populations (113 +/- 74 IU/ml for SOD and 1.17 +/- 0.52 IU/ml for XO) in samples from normozoospermic ejaculates. Sperm progressive motility was positively correlated with SOD activity in seminal plasma of corresponding ejaculates (P < 0.05) and negatively with XO activity (P < 0.001). An 'oxido-sensitive' index was defined as the SOD/XO ratio and was found to be inversely related to sperm progressive motility samples (P < 0.01). Analysing this index among all tested samples of semen including those with pathological spermiograms, as well as normospermic (N) samples we found statistically significant (elevated) differences in oligoasthenoteratospermia (OAT) in comparison with N (P <0.05); OAT samples were also significantly different from oligospermic (O) and oligoteratospermic (OT) samples (P < 0.05). This suggests that the 'oxido-sensitive' index of seminal plasma may be a simple diagnostic factor, useful in the determination of male infertility.

Clinical Enzyme Tests↗
PubMed1996

[Immunologic response against spermatozoa in the human genital tract].

Spermatozoa are immunogenic and may induce an immune response in reproductive compartments. There have been controversies concerning the effect of naturally existing antisperm antibodies on fertility of males and females. It is known, that men or women suffering from unexplained infertility have often antisperm antibodies in their urogenital secretions and/or serum. Systemic and local immune responses to sperm antigens differ between each other and they may differently affect the fertilisation process. Physiologically, females are protected from an immune response against "foreign", spermatozoal antigens. Nevertheless under local pathological conditions antisperm antibodies can be induced and may interfere with fertilisation.

Adult↗
PubMed1995

Antiinflammatory effects of NADPH oxidase inhibitors.

Proinflammatory cytokines prime the membrane-bound NADPH oxidase of neutrophils and monocytes of mice suffering from experimental arthritis so as to attain an activated state, which, upon a second stimulus, releases 6-fold increased levels of reactive oxygen species (ROS) than do unprimed phagocytes. Enhanced NADPH oxidase activity deregulates ROS-dependent signal transduction pathways of inflammation, which play a crucial role in the pathogenesis of arthritis. The antiarthritic reactivity of two inhibitors of NADPH oxidase, diphenylene iodoniumchloride (DPI) and staurosporine, was tested in male DBA/1 x B10A(4R) hybrid mice suffering from potassium peroxochromate arthritis. Daily doses of 2.8 mumol/kg of DPI or 30 nmol/kg of staurosporine sufficed to inhibit the arthritis by 50%. A complete inhibition was obtained with 10 mumol/kg of DPI, and 100 nmol/kg of staurosporine suppressed the arthritis by 85%. The onset, progression, and remission of arthritis correlated to both the activity of phagocytic NADPH oxidase (r = 0.750) and to overt disease symptoms as judged by the arthritis index. Our data support the hypothesis that oxidative stress plays a pivotal role in the pathology of arthritis, which can be therapeutically targeted by NADPH oxidase inhibitors.

Alkaloids↗
PubMed1995

Modulation of inflammatory arthritis by inhibition of poly(ADP ribose) polymerase.

Poly(ADPR) polymerase (PARP; EC 2.4.2.30) is a nuclear enzyme, which, when activated by oxygen- and nitrogen-radical-induced DNA strand breaks, transfers ADP ribose units to nuclear proteins and initiates apoptosis by depletion of cellular NAD and ATP pools. The present study investigates whether the oxidative stress-dependent activation of PARP plays a role in the etiopathogenesis of arthritis. The antiarthritic reactivity of the biogenic PARP inhibitor nicotinamide was tested in DBA/1 x B10A(4R) mice suffering from potassium peroxochromate-induced arthritis. Daily doses of 4 mmol/kg of NA suppressed the arthritis by 35% and inhibited the phagocytic generation of reactive oxygen species, which increases sixfold during the development of arthritis. The onset, progression, and remission of arthritis correlated positively to the phorbolester-activated respiratory burst of neutrophils and monocytes, and a dose-dependent inhibition of NADPH oxidase activity was determined with human phagocytes. Our data support the hypothesis that oxidative stress-induced alterations in cellular signal transduction pathways play a pivotal role in the development of arthritis, which can be suppressed by the simultaneous inhibition of poly(ADPR) polymerase and NADPH oxidase.

Animals↗
PubMed1995

Analysis of mRNA expression for interleukin-1 genes on human testicular cells.

We have investigated mRNA expression for IL-1 alpha and IL-1 beta gene on fractionated human testicular cells. Using RT-PCR and Northern blot hybridization technique we detected the presence of IL-1 alpha transcripts, predominantly in the intratubular compartment of the testis, comprising gametogenic and Sertoli cells. We were also able to detect mRNA for IL-1 alpha on the testicular interstitium, but at significantly lower levels. The intertubular compartment of the testis, mainly consisting of macrophages and Leydig cells, appeared however, to be a site for IL-1 beta gene expression. Our experimental data confirm previous results obtained in animal models indicating that the testis is capable of producing interleukin-1 under physiological conditions. Testicular IL-1 may function as a tissue-specific factor modulating both spermato- and steroidogenic activity of human testis.

Base Sequence↗
PubMed1995

Hepatoprotective reactivity of a copper-di-Schiffbase active centre analogue of Cu2Zn2 superoxide dismutase.

1. The anti-inflammatory and hepatoprotective efficacy of CuPu(Py)2 ((N,N'-bis(2-pyridyl-methylene)-1,4-butanediamine) (N,N',N",N")-Cu2+), a serum-stable, copper-di-Schiffbase active centre analogue of Cu2Zn2 superoxide dismutase was tested in male NMNR mice suffering from endotoxin/galactosamine-induced hepatitis. 2. Parameters including the activities of serum transaminases and sorbitol-dehydrogenase as well as the levels of reactive oxygen and nitrogen intermediates which were used to quantify the disease activity. 3. A dose-dependent inhibition of hepatic enzyme release was noted in the presence of 0.1-10 mg/kg of CuPu(Py)2. 4. The release of transaminases from damaged liver cells was reduced by 68% and paralleled the reduction of serum levels of nitric oxides. 5. Elevated levels of reactive oxygen species were normalized to those healthy controls. 6. The copper-free apochelate Pu(Py)2, which is unable to dismutate superoxide, did not display any anti-inflammatory reactivity.

Alanine Transaminase↗
PubMed1995

Induction of arthritis in mice and rats by potassium peroxochromate and assessment of disease activity by whole blood chemiluminescence and 99mpertechnetate-imaging.

Arthritis develops in DBA/1xB10A(4R) mice and Wistar rats upon intraplantar injection of potassium peroxochromate (K3CrO8), and is here quantified by whole blood chemiluminescence (CL) and 99mpertechnetate-imaging (99mTcO4-), and related to overt disease symptoms (the arthritis index). During the aqueous decay of K3CrO8 to chromate (VI), the chromium(V)-bound oxygen is released as superoxide, hydroxyl radicals, singlet oxygen and hydrogen peroxide, the same reactants, which are produced by activated phagocytes during inflammation. Reactive oxygen species (ROS) trigger the breakdown of the sulfhydryl-dependent antioxidant defence system and induce the nuclear factor kappa B-dependent expression of pro-inflammatory cytokines, which prime phagocytic NADPH oxidases to the enhanced production of ROS. During both the acute inflammatory response and the onset of the secondary response in non-injected paws, the phorbolester-stimulated ROS production of phagocytes was significantly enhanced (p < 0.001) and correlated well to the arthritis index (r = 0.797) and the uptake of 99mTcO4- into inflamed joints. Chromate(VI), formed during the decay of K3CrO8, contributes to the progression of arthritis by inhibition of glutathione reductase, thereby increasing intracellular H2O2 concentrations. In addition, Cr(VI) reduced to Cr(V) by ascorbate, catalyzes hydroxyl radical production in the presence of hydrogen peroxide. A stable loop forms, in which ROS, continuously produced by Cr(VI)/Cr(V) redox-cycling, drive the primary response into chronic self-perpetuating inflammation. We see the main application of K3CrO8-induced arthritis and its assessment by both 99mTcO4- imaging and chemiluminescent immunosensoring of phagocytic activity in unseparated blood as for the rapid screening of novel anti-rheumatic drugs and treatments.

Animals↗
PubMed1995

Carbohydrate moieties on sperm surface: physiological relevance.

OBJECTIVES: To study the cross-reactions between mouse monoclonal antisperm antibodies and somatic cells or bacteria, to identify the antigenic determinants responsible for such cross-reactions, and to correlate between the antibody function and determinant recognition. DESIGN: Activities of monoclonal antibodies (mAbs) were characterized by immunosorbent assay of RIA technique; sperm epitopic characterization was performed in lectin-blocking and sugar competitive assays and correlated with functional assays. SETTING: Procedures were performed in a university laboratory. RESULTS: The extensive cross-reactivity between antigenic determinants of sperm, erythrocytes, and bacteria (but not bacterial deglycosylated lipopolysaccharides) was observed. The analytic procedures indicated predominant mAb reactions to carbohydrates such as fucose, galactose, mannose, N-acetylglucosamine and N-acetylgalactosamine. Approximately half of the 30 tested mAbs interfered in the functional assays, that is, sperm agglutination, immobilization, and zona-free penetration. CONCLUSIONS: Sperm carbohydrates seem to induce antibody reactions to common antigenic determinant(s) present on gametes, somatic cells, and infectious agents. Thus molecular mimicry between bacteria and sperm can be a major factor inducing antisperm immunological reactions. Obtained antisperm mAbs, reacting to glycosylated epitopes, presented very strong properties in sperm agglutination and/or immobilization. This did not correlate with inhibiting properties of some antibodies in xenogeneic zona-free penetration test, that is, this assay possibly is not based on oligosaccharide mediation.

Animals↗
PubMed1995

[Immunologic reactions in the male and female reproductive system].

This review describes presence and significance of antisperm antibodies (ASA) in male and female reproductive compartments. We described their relevance to infertility in humans. A structure and function of cervical mucus as the environment of females defensive reactions against all the invasive factors is outlined. There are also briefly characterized techniques of ASA detection.

Agglutination Tests↗
PubMed1995

X- and Y-chromosome-bearing sperm selection and detection methods. A review.

The ability to preselect the sex of human offspring would have a significant impact on the preventing of sex chromosome-linked diseases and on human reproduction. Numerous methods have been reviewed including: Sephadex gel filtration, centrifugation in density gradients, separation in albumin gradient, laminar flow, counterstreaming centrifugation, free-flow electrophoresis, galvanic method, two-phase system separation, flow cytometry, immunological exclusion. None of these methods seems to be ideal. Results obtained with the use of the most of them are contradictory, the others still need reliable confirmation. Part of the controversy related to the efficacy of methods aimed at sex selection may be due to the lack of appropriate and widely used techniques of X- and Y-chromosome bearing sperm detection. Fluorescent in situ hybridization appears to be the most suitable among the sperm sex chromosome detection techniques described in this review. Others, like quinacrine staining method, human sperm/hamster oocyte penetration assay, microinterferometry, cytometry, and polymerase chain reaction exhibit more or less disadvantageous features, which limit their application.

Cell Separation↗
PubMed1994

Analysis of mRNA for class I HLA on human gametogenic cells.

We have studied mRNA expression for Class I HLA (human leukocyte antigen) on male germ cells by amplification of gene fragments in PCR technique and by Northern hybridization. RNA was extracted from fractionated gametogenic cells (isolated from testis) and reversely transcribed. Then, cDNA was amplified for specific HLA sequence (HLA, -A, -B, -C). The specificity of this product was confirmed in "nested" PCR of 400 bp gene fragment coding for alpha 2 domain, alpha 3 domain, and the transmembrane portion of Class I HLA. The results indicate minimal expression of classical Class I HLA on gametogenic cells. Northern hybridization with 669 bp cDNA fragment (spanning for alpha 3 domain, transmembrane, cytoplasmic, and 3' untranslated region) resulted in a low intensity signal from gametogenic cell fractions and confirmed our findings obtained by PCR. The minimal expression of classical HLA antigens may create a neutral cover for the male reproductive system, thereby preventing an immunological response during germ cell differentiation.

Blotting, Northern↗