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Biomedical subjects

M Kurpisz

Publications and source records attributed to M Kurpisz.

At least 19 recordsLinked to original sources

Interleukin-1 superfamily genes expression in normal or impaired human spermatogenesis.

Interleukin-1 (IL-1) is a pleiotropic cytokine that may play a role in contributing to the specific immune environment of mammalian testis and in regulating cell differentiation. We have determined the transcription activity of the IL-1 gene family (using real-time polymerase chain reaction (PCR)) in two main functional testicular compartments (interstitial and intratubular ones), and in tissue homogenates obtained from patients with fertility disorders (spermatogenic arrest and testicular tumors). We observed the prominent expression of gene coding for IL-1 receptor antagonist (IL-1RA) in a purified fraction of gametogenic cells (normal gonad). Caspase-1 (ICE - IL-1beta-converting enzyme) was highly expressed (on mRNA level) in interstitial compartments as well in testicular tumors (immune enhancement?). In addition we found, that the activity of IL-1RA gene decreased along spermatogenic alteration in an inversely related manner with IL-1alpha (from normal gonad through spermatogenic arrest to Sertoli cell only syndrome). Therefore, the quotient value of IL-1alpha/IL-1RA could potentially serve as the diagnostic molecular probe for spermatogenesis assessment. The precise level of mRNA for IL-1-IL-18 cytokines and their receptors, and specifically of the receptor antagonist in immune privileged gonad, could be one of the main factors responsible for maintaining testicular homeostasis, thus enabling generation of the mature spermatozoa.

Caspase 1↗

Bone marrow stem cell imaging after intracoronary administration.

Although feasibility and safety of autologous stem cells administration to the post-infarction heart has been proven it is not known what proportion of cells effectively do home at the damaged site. Therefore, we have labeled autologous bone marrow cells (ABMC's) by radioactive Indium and single photon emission computed tomography (SPECT) tissue distribution has been analyzed. It was detected that up to 10% of the cells were retained within the myocardium while their majority migrated or has been anchored at the spleen and liver. Comparing the number of homed cells to the total number of cells delivered one may postulate the indirect role for few hundred thousands ABMC's at heart regeneration.

Aged↗

Differential effect of IFN-tau on proliferation and distribution of lymphocyte subsets in one-way mixed lymphocyte reaction in cows and heifers.

IFN-tau is a signaling protein secreted by the bovine conceptus during the peri-implantation period and responsible for pregnancy recognition. Its main role is the prevention of pulsatile release of luteolytic PGF2alpha, but it also exerts immunomodulatory activities characteristic for other type I interferons. The aim of the study was to examine the effect of IFN-tau on the proliferation and distribution of peripheral blood lymphocyte subsets during one-way mixed lymphocyte reaction (MLR) in cows and heifers. IFN-tau inhibited the proliferative response of lymphocytes in MLR both in cows and heifers in a dose-dependent manner, but cow lymphocytes were less susceptible than those ones from heifers. It was also showed that IFN-tau differentially changed lymphocyte subsets distribution in MLR in cows and heifers. In cows, the relative percentage of CD8(+) cells after MRL in the presence of IFN-tau was significantly lower than in heifers. Differential effect of rIFN-tau on proliferation and lymphocyte subsets distribution in a one-way MRL in cows and heifers indicated that the age of the mother is an important factor in immunomodulatory effect towards developing bovine embryo.

Analysis of Variance↗

Bacterial infection and semen quality.

We have analyzed two infertile male cohorts with (n=39) and without genital tract infection (n=14) comparing their selected seminological parameters with healthy controls (n=30). Genital tract infection (GTI) has been defined by the presence of leukocytes and pathological bacterial strains identified with Bio-Merieux tests. We have found statistically significant deteriorated semen volume, sperm concentration, motility, morphology and vitality in ejaculated samples of patients with genital tract infection in comparison to healthy controls. Statistically significant negative influence towards sperm reproductive potential has been revealed in case of Escherichia coli, Ureaplasma urealyticum and Staphylococcus aureus.

Adult↗

Predictive value of selected sperm parameters for classical in vitro fertilization procedure of oocyte fertilization.

A proportion of fertilized oocytes during classical in vitro fertilization (IVF) procedure was analysed depending on the following factors: number of mature oocytes, seminological criteria such as sperm morphology in raw semen and after its selection in a density gradient (six structural defects of a male gamete were taken into consideration), sperm concentration, motility parameters according to World Health Organization criteria and the functional tests: hypo-osmotic swelling assay and acrosomal reaction induced by calcium ionophore. Evaluation of DNA content in sperm by image cytometry and determination of malonyldialdehydes in seminal plasma were also performed. Seventy-nine semen samples from patients undergoing IVF were assessed. Apart from significant correlations obtained for selected semen parameters and proportion of fertilized eggs, logistic regression analysis showed that the best predictive factors for oocyte fertilization were normal morphology of sperm before and after gradient selection, grade B and C of sperm movement in raw semen, and DNA content after density gradient centrifugation, which all accounted for 76.7% of fertilization predictive value.

Fertilization in Vitro↗

Complex nature of the human antisperm antibody response in SCID mice.

Human peripheral blood mononuclear (PBMs) cells were introduced into the peritoneal cavity of severely-combined immunodeficient (SCID) mice in concentrations of 2.5-4.0 x 10(7) cells per mouse. Whole mononuclear cell suspensions were used either unstimulated or following primary in vitro culture with human spermatozoa. In some experiments, immunodepletion of CD8(+) cells was carried out prior to grafting. Lymphocytes were obtained from nonsensitized (to antigen) human subjects or from individuals who were primed in vivo (vasectomized individuals in case of sperm antigens). An enzyme-linked immunosorbent assay was employed to assess total human immunoglobulin (G or M) levels as well as the specificity of the antibodies generated. We have been successful by generating primary and secondary immune responses with 'naïve' human lymphocytes, challenged with chlamydia or ovalbumin but without adjuvant or CD8(+) immunodepletion; however, we were unable to induce specific antibodies to spermatozoa under this regime in SCID male mice. We then employed female SCID mice, treated with sperm antigen extracts (glycosylated or deglycosylated) encapsulated in liposomes and human lymphocytes obtained from 'naïve' or pre-sensitized in vivo subjects. It was found that the most pronounced humoral response to sperm antigens was obtained with deglycosylated antigens and PBMs from vasectomized (in vivo pre-primed to spermatozoa) individuals. A presented SCID mice model can be helpful at understanding of antisperm antibody development and the molecular nature of generated antibodies to modified sperm antigenic entities.

Adult↗

Quantitative mRNA analysis of IL-1 gene system in human testis.

PROBLEM: There is a growing body of evidence that interleukins exhibit modulatory activity on development of reproductive cells. In this context, there appears to be a role for IL-1, which is also produced in human testis. We have analysed transcripts of IL-1 gene system (IL-1alpha, IL-1beta, IL-1RI, IL-1RII and IL-1RA) to evaluate the possible link between the level of gene(s) transcription and their function. METHOD OF STUDY: To determine the activity of gene transcription, a quantitative PCR with isotopic and/or nonisotopic detection was applied. RESULTS AND CONCLUSIONS: We have detected differential expression of IL-1alpha and IL-1beta genes in separate functional compartments of a male gonad. A strong expression of IL-1alpha gene in an intratubular cell fraction was shown, while the IL-1beta expression seemed to be dominant in extratubular compartment of the male gonad. Abundant amounts of IL-1RA mRNA in gametogenic cells fraction slightly higher than in interstitium have also been found. IL-1RA is the most important regulatory molecule in IL-1 system, which down-regulates activity of both interleukins. Looking more closely at gene(s) differential expression it appears that IL-1alpha can be preferentially down-regulated by IL-1RA gene in intratubular fraction while the IL-1beta, through the "false" IL-1RII receptor in the interstitium. Genes coding for both receptors (IL-1RI and IL-1RII) showed, however, relatively low levels of transcription in both studied compartments. IL-1 genes system creates a complex intragonadal environment and the function of these genes is reflected by their respective distribution in the two main functional compartments of the testis.

Base Sequence↗

Antizona and antisperm antibodies in women with endometriosis and/or infertility.

OBJECTIVE: To measure the levels of antigamete antibodies in serum and peritoneal fluid of women with endometriosis and/or infertility. DESIGN: Antibody activity against human sperm and porcine oocytes was analyzed in selected subgroups of women. SETTING: Clinic of reproduction. PATIENT(S): Women with endometriosis and/or infertility. INTERVENTION(S): No treatment was implemented before peritoneal fluid and blood sample collection. MAIN OUTCOME MEASURE(S): Quantitative ELISA. RESULT(S): Four groups of women (n = 98) were analyzed for the presence of antizona and antisperm antibodies: infertile with endometriosis (n = 30), idiopathic infertility (n = 28), fertile with endometriosis (n = 20), and healthy fertile controls (n = 20). Antibodies were analyzed simultaneously in serum and peritoneal fluid. No statistically significant differences in antibody levels were detected in serum samples among the analyzed groups. The median values for antizona and antisperm antibodies in peritoneal fluid were significantly higher in women with idiopathic infertility than in the control group. In women with unexplained infertility, a high degree of correlation (Spearman) was found between the presence of antizona antibodies in peritoneal fluid and serum (r = 0.579). A positive predictive value of 80% was calculated for the presence of antizona antibodies (>5 ng/oocyte) in the peritoneal fluid of patients with infertility. CONCLUSION(S): Antizona antibodies locally produced in the peritoneal fluid have diagnostic value for infertility status; however, they cannot be treated as a marker or prognostic factor for minimal endometriosis and/or its treatment.

Adult↗

The effect of Ureaplasma diversum activated mononuclear leukocytes on the development and interferon-tau production by bovine IVF-derived embryos.

Ureaplasma diversum is an opportunistic pathogen of the bovine genital tract causing herd outbreaks of granular vulvitis, abortion and infertility. Early embryonic death probably contributes to reduction of the reproductive performance in cows, however, pathogenesis of the disease remains obscure. The aim of the study was to examine whether activation of mononuclear leukocytes by U. diversum may affect embryo development and IFN-tau production. Bovine peripheral blood mononuclear leukocytes were cultured with U.diversum antigen for 24 h. The levels of IL-1, TNF-alpha, NO and GM-CSF in the cell culture supernatants were measured. IVF-derived embryos were cultured in the presence of supernatants from activated leukocytes. The development of embryos until day 6 postinsemination and the rate of morulae/blastocysts were determined. IFN-tau production in supernatants of cultured embryos was examined by inhibition of a virally-induced cytopathic effect. The results showed that U. diversum stimulated mononuclear leukocyte production of IL-1, TNF-alpha and NO. Supernatants from U. diversum-activated cells did not impair the rates of the embryo development and blastocyst formation. The products of activated leukocytes increased the IFN-tau production by cultured blastocysts. This suggest that U. diversum infection provides leukocyte-mediated signals for developing embryos for generation of additional production of cytokine - an important component of innate immunity.

Animals↗

Chromosomal anomalies in human gametes and pre-implantation embryos, and their potential effect on reproduction.

This paper reviews the latest data on chromosomal abnormalities in human gametes and embryos. A close relationship between such anomalies and reproduction failure in humans has been postulated, thereby underscoring the importance of ongoing studies into the mechanisms leading to anomalies. Until recently, knowledge of chromosomal anomalies in human gametes and embryos has been limited. Newly developed strategies (in vitro fertilization combined with micromanipulation techniques followed by multicolour fluorescence in situ hybridization, and PCR analyses) allow precise investigation of this problem. This review of the available information on the etiology of chromosomal anomalies indicates that some of the genetic anomalies in human gametes and early embryos result in reproductive failure.

Chromosome Aberrations↗

Idiopathic infertility in married couples in the light of cytogenetic analysis and sperm penetration assay.

We have selected 47 couples with unexplained infertility in order to analyse a possible link between sperm dysfunction studied in males in in vitro conditions and karyotype analysis of somatic cells. In order to identify so called "idiopathically infertile" couples we had to exclude any change in reproductive organs in both partners or in spermiogram which would qualify any of spouses into known category of infertility. We have revealed chromosome aberrations (translocations and marker chromosomes) in 19% of infertile males and in 6% of infertile females. Idiopathically infertile males had an overall decreased ability of sperm function (measured by proportion of penetrated hamster oocytes by human sperm) in comparison to fertile controls, however, still well placed within physiological range of values. Only sperm from a patient with identified translocation was clearly below the normal level of penetration (20% of penetrated oocytes), however, also the patients with revealed chromosome variant polymorphisms presented statistically lower values of penetration in comparison to fertile controls (39% vs 57%, p<0.05). On the contrary, patients with marker chromosomes did not exhibit affected sperm function. It can be speculated that only particular chromosome aberration in group of idiopathically infertile males may affect sperm functional capability (measured in vitro), however, the intragonadal genetic analysis has to be recommended in order to confirm such a causative link.

Adult↗

[Peroxidation components of sperm lipid membranes in male infertility].

We have studied the lipid peroxidation product (malondiadehyde-MDA) levels after thiobarbituric acid reaction, spectrophotometrically and by a high-performance liquid chromatography (HPLC) with UV detection in seminal plasma as well as in the cell fraction. Semen samples were obtained from healthy volunteers and infertile males. Ejaculates were previously classified into studied subgroups according to standard andrological criteria (sperm number, motility, morphology) and defined as: normozoospermia (K), azoospermia (Az), teratozoospermia (T), asthenoteratozoospermia (AT), oligoasthenoteratozoospermia (OAT) and idiopathic infertility (NIF). MDA is a good marker for lipid peroxidation. There were found elevated MDA concentrations (determined by HPLC) in seminal plasma of the all analysed pathological semen samples, especially in patients with OAT. The more pronounced differences between healthy controls and NIF patients were observed in intracellular compartment. The lipid peroxidation of rich in unsaturated fatty acids sperm membranes is considered to be one of the most important effects from ROS-induced cell damage what may lead to persistent infertility.

Adult↗

Sperm antigens recognized by antisperm antibodies present in sera of infertile adults and prepubertal boys with testicular failure.

Immunoblotting of a repertoire of sperm antigens reacting with antisperm antibodies present in sera of infertile adults and prepubertal boys with testicular failure was performed. In the subgroups selected for this study, 55% of examined infertile women, 65% of infertile men and 64% of prepubertal boys with gonadal failure gave positive results by Western blotting with extracted sperm antigens. Sperm antigens with molecular weights of 57, 58, 62, 63 and 66 kDa were the most immunodominant entities recognized by antisperm antibodies from prepubertal boys. No positive reactions were detected by Western blotting in a control population of fertile adults, whereas in a group of prepubertal healthy boys only one sample revealed reactivity against sperm antigens of 58 and 70 kDa.

Adolescent↗

[Molecular analysis of protein 4.1 gene in teratozoospermic and azoospermic patients].

In normal sperm, the 135 kD isoform of protein 4.1 is replaced by the 80 kD variant. Transcript for protein 4.1 loses the 'upstream' initiation codon by a stage-dependent alternative splicing and in mature sperm only 'downstream' initiation codon is active. A mutation in the 'downstream' initiation codon may be a reason for sperm differentiation arrest (azoospermia) or can be associated with the presence of amorphous spermatozoa in ejaculate (teratozoospermia). The aim of the study was the molecular analysis of gene coding for the protein 4.1, carrying a 'downstream' translation initiation codon. We have screened DNA samples obtained from azoospermic (blood) and teratozoospermic (spermatozoa) patients using PCR amplification of gene fragment, AUG containing exon with subsequent digestion (NlaIII) of CATG sequence. The absence of a cleavage site for this restriction enzyme would suggest the presence of mutation in the AUG codon. Analysis of DNA samples obtained from both azoospermic and teratozoospermic patients did not reveal any changes in the 'downstream' translation initiation codon. We concluded therefore that observed by others a defective expression of protein 4.1 in amorphous sperm cells is probably due to the other factor(s) than mutation in the 'downstream' translation initiation codon.

Codon↗

[Reconstruction of humoral response to sperm antigens in scid mice].

OBJECTIVE AND DESIGN: Production of specific human antisperm antibodies by using human-SCID mice model with deposited peripheral blood lymphocytes. MATERIALS AND METHODS: Human peripheral blood lymphocytes (PBL's; CD8(+)-negative cell fraction) were grafted to the peritoneal cavity of severely-combined immunodeficient (SCID) mice at concentration of 20-35 x 10(6) cells per mouse. Lymphocytes were obtained from non-sensitized individual (to sperm antigen) and from in vivo primed males (vasectomized). Two sets of experiments were carried out, with 'native' (glycosylated) and enzymatically deglycosylated sperm antigenic extracts. In all applied variants, sperm antigens were administered with Complete and then with Incomplete Freund adjuvant to improve an immune response. RESULTS AND CONCLUSION: This approach allowed us to obtain better pronounced humoral antisperm response, specific to sperm deglycosylated antigens when PBL's were obtained from individuals in vivo sensitized to sperm (after vasectomy).

Animals↗

[Molecular basis of teratozoospermia].

Teratozoospermia is considered to be one of the most challenging phenomenon of male infertility at proper diagnosis and therapy. In this report, we described controversies regarding current methods of evaluation of sperm morphology and predictive value of this sperm feature in connection to assisted reproduction and/or functional sperm tests in vitro. It is indicated possible molecular background of teratozoospermia in respect to revealed chromosome aberrations and particular genes of which mutations may affect a sperm morphology and function. The own experience in research conducted in this field is also underlined.

DNA Mutational Analysis↗