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Biomedical subjects

A Sobek

Publications and source records attributed to A Sobek.

At least 19 recordsLinked to original sources

Observations of the PCB distribution within and in-between ice, snow, ice-rafted debris, ice-interstitial water, and seawater in the Barents Sea marginal ice zone and the North Pole area.

To evaluate the two hypotheses of locally elevated exposure of persistent organic pollutants (POPs) in ice-associated microenvironments and ice as a key carrier for long-range transport of POPs to the Arctic marginal ice zone (MIZ), dissolved and particulate polychlorinated biphenyls (PCBs) were analyzed in ice, snow, ice-interstitial water (IIW), seawater in the melt layer underlying the ice, and in ice-rafted sediment (IRS) from the Barents Sea MIZ to the high Arctic in the summer of 2001. Ultra-clean sampling equipment and protocols were specially developed for this expedition, including construction of a permanent clean room facility and a stainless steel seawater intake system on the I/B ODEN as well as two mobile 370 l ice-melting systems. Similar concentrations were found in several ice-associated compartments. For instance, the concentration of one of the most abundant congeners, PCB 52, was typically on the order of 0.1-0.3 pg l(-1) in the dissolved (melted) phase of the ice, snow, IIW, and underlying seawater while its particulate organic-carbon (POC) normalized concentrations were around 1-3 ng gPOC(-1) in the ice, snow, IIW, and IRS. The solid-water distribution of PCBs in ice was well correlated with and predictable from K(ow) (ice log K(oc)-log K(ow) regressions: p<0.05, r2=0.78-0.98, n=9), indicating near-equilibrium partitioning of PCBs within each local ice system. These results do generally not evidence the existence of physical microenvironments with locally elevated POP exposures. However, there were some indications that the ice-associated system had harbored local environments with higher exposure levels earlier/before the melting/vegetative season, as a few samples had PCB concentrations elevated by factors of 5-10 relative to the typical values, and the elevated levels were predominantly found at the station where melting had putatively progressed the least. The very low PCB concentrations and absence of any significant concentration gradients, both in-between different matrices and over the Eurasian Arctic basin scale, suggest that ice is not an important long-range transport purveyor of POPs to the Arctic MIZ ecosystem.

Journal Article↗

[Importance of genetic testing in couples with reproductive disorders].

OBJECTIVE: To determine the prevalence of chromosomal aberrations in infertile couples undergoing in vitro fertilization (IVF). DESIGN: Cytogenetic analysis of peripheral blood lymphocytes in the group of patients undergoing IVF. Detection of chromosomal aberrations in the fetuses after IVF. SETTING: Department of Medical Genetics and Fetal Medicine, Medical Faculty, Palacký University and the University Hospital, Olomouc. METHODS: Cultivation of peripheral blood lymphocytes or fibroblasts of amniotic fluid. Using fluorescent in situ hybridization in cases of mosaicism. RESULTS: Out of 638 patients undergoing treatment for male or female infertility, 595 had normal karyotype and 43 (6.8%) had abnormal karyotype. There were detected 9 (1.4%) cases of balanced chromosomal rearrangements, 2 (0.31%) cases of deletion of Y chromosome, 2 (0.31%) cases of inversion, 2 (0.31%) cases of marker chromosome, 5 (0.78%) cases of gonosomal aneuploidy (47,XXY) and 23 (3.65%) cases of gonosomal mosaicism--out of the 22 (3.5%) cases of low-level mosaicism. In the small group of pregnant patients after IVF investigated for the risk of genetic disorders included in our study (n = 60) the frequency of chromosomal abnormalities was 9 (15%). CONCLUSIONS: Our data show that a high number of infertile couples is affected by chromosomal aberrations which occur more frequently in females than in males. It is caused by high frequency of low-level gonosomal mosaicism in the group of infertile women. Chromosomal analyses are highly recommended before each IVF procedure.

Chromosome Aberrations↗

[Chromosome Y microdeletions in Czech men with severe reproductive disorders].

BACKGROUND: The Y chromosome microdeletions belong to the frequent genetical causes of male infertility. The aim of our study was to introduce reliable molecular genetic diagnosis of Y chromosomal microdeletions and to determine the prevalence of Y chromosomal microdeletions in Czech males with serious reproductive disorders. METHODS AND RESULTS: The Y chromosome microdeletions were screened in 198 Czech men with serious reproductive disorders with decreased sperm count. The Y chromosome microdeletions were disclosed in 8/198 (4.0%) examined males. The AZFc deletion type was revealed in 62.5% (5/8) and the combined AZFc + b microdeletion in 37.5% (3/8) of cases. Neither isolated AZFb nor AZFa microdeletion were found in any subject of the investigated group. CONCLUSIONS: Incidence of individual types of Y chromosomal microdeletions in Czech males with serious reproductive disorders was assessed. The standardised molecular genetic diagnosis of Y chromosomal microdeletions was introduced into the practice.

Chromosomes, Human, Y↗

Biochemical findings in the ejaculate of men with non-obstructive azoospermia.

In men with non-obstructive azoospermia, testicle biopsy can provide isolated sperm which can be used for fertilization of an oocyte. The male seminal plasma was examined for adequate biochemical parameters and then tested as potential diagnostic parameters for prediction of sperm presence in non-obstructive azoospermia. Sperm was obtained by preparation of the testicular tissue. Biochemical parameters in men with found sperm were compared to findings obtained from men without sperm in the testicular tissue. The following parameters were analysed: fructose, lactate dehydrogenase, total acid phosphatase, Zn, K, Na, Cl, pH. No statistically significant differences were found in both groups under study. Biochemical examination of the ejaculate cannot provide any prediction of sperm retrieval in non-obstructive azoospermia.

Humans↗

Infertility treatment of men with non-obstructive azoospermia.

Male infertility can be treated by several methods with varying degree of success. We present evidence that "open" testicular biopsy is favorable for men suffering from non-obstructive azoospermia (NOA). Moreover, any NOA patient may be subjected to this treatment even though his past histopathological examinations suggest that it is likely no sperm will be found in the testicular tissue. Thus, we recommend the testicular sperm extraction (TESE) procedure for NOA patients.

Female↗

[Treatment of male infertility after oncologic therapy: preventive autologous sperm preservation versus testicular biopsy and oocyte fertilization with testicular spermatozoa].

The prognosis of oncological therapy of some malignancies is favourable. Unfortunately the majority of young patients remain sterile. Sperm cryopreserved before treatment was successfully used in IVF-ICSI (in vitro fertilization-intracytoplasmic sperm injection) cycles. Testicular biopsy (TESE) was performed after successful oncological treatment in three men without previous sperm cryopreservation. No spermatozoa were found for oocyte fertilization. TESE could not ensure sperm recovery in all patients after oncological treatment. Sperm cryopreservation should be highly recommended to all patients before any treatment is offered.

Cryopreservation↗

[Treatment of infertility in non-obstructive azoospermia using the TESE (testicular sperm extraction) method--clinical study].

Testicular sperm extraction (TESE) was performed in 27 men in 30 cycles. All men were examined for genetics, serum hormonal status, biochemical status of semen samples. All men were examined by an urologist. No prognostic evaluation able to provide information about the prognosis of TESE procedure was found. Even a high FSH level, testicular hypotrophy or previous histological examination cannot exclude any patient from testicular biopsy.

Adult↗

[Treatment of infertility in nonobstructive azoospermia using the TESE method (testicular sperm extraction)--clinical results].

Sperm was found in 56.6% of TESE cycles in 27 men with non-obstructive azoospermia. Using testicular sperm, 30% oocytes were fertilized. Embryos were transferred in 11 cycles. Twins were delivered in the 34th week of pregnancy. Two further pregnancies are ongoing. The pregnancy rate was 27% per ET. Testicular biopsy in all azoospermic men is recommended in special IVF centers not only for histological examination but also for ICSI procedure with testicular sperm which could be provided at the same time.

Biopsy, Needle↗

Involvement of proteasomal subunits zeta and iota in RNA degradation.

We have identified two distinct subunits of 20 S proteasomes that are associated with RNase activity. Proteasome subunits zeta and iota, eluted from two-dimensional Western blots, hydrolysed tobacco mosaic virus RNA, whereas none of the other subunits degraded this substrate under the same conditions. Additionally, proteasomes were dissociated by 6 M urea, and subunit zeta, containing the highest RNase activity, was isolated by anion-exchange chromatography and gel filtration. Purified subunit zeta migrated as a single spot on two-dimensional PAGE with a molecular mass of approx. 28 kDa. Addition of anti-(subunit zeta) antibodies led to the co-precipitation of this proteasome subunit and nuclease activity. This is the first evidence that proteasomal alpha-type subunits are associated with an enzymic activity, and our results provide further evidence that proteasomes may be involved in cellular RNA metabolism.

Animals↗

Subunit arrangement in the human 20S proteasome.

In human 20S proteasomes two copies of each of seven different alpha-type and seven different beta-type subunits are assembled to form a stack of four seven-membered rings, giving the general structure alpha(1-7), beta(1-7), beta(1-7), alpha(1-7). By means of immunoelectron microscopy and chemical crosslinking of neighboring subunits, we have determined the positions of the individual subunits in the proteasome. The topography shows that for the trypsin-like, the chymotrypsin-like, and the postglutamyl cleaving activities, the pairs of beta type subunits, which are thought to form active sites, are nearest neighbors.

Amino Acid Sequence↗

[The current role of intracytoplasmic single sperm injection (ICSI) in assisted reproduction].

Intracytoplasmic sperm injection (ICSI) plays a unique role in the treatment of male infertility. ICSI results are not influenced by either sperm number, motility or sperm morphology. In our group we studied 90 cycles in which conventional IVF and ICSI were performed on sibling oocytes in couples with borderline semen analysis (more than 50,000 sperm/oocyte). A higher number of A quality embryos was found in the ICSI oocyte group (40%/32%). In 38% of IVF cycles no embryos were found, whereas with ICSI the absence decreased to only 12%. In only 4% of the cycles, the IVF method was more successful than that of ICSI treatment. In only 7% of our combined group there were no embryos for transfer. The positive influence of ICSI on our centre results -32% pregnancy rate/ET and an 18% baby take home rate--are discussed. In our retrospective study, ICSI is more successful than IVF alone and a combination of IVF and ICSI achieves the best results in the borderline sperm analysis group. An increased use of the ICSI technique in assisted reproduction can be expected in future.

Female↗

[The first pregnancy in the Czech Republic achieved by oocyte fertilization using ICSI--intracytoplasmic sperm injection].

ICSI--an intracytoplasmic sperm injection can fertilize an oocyte in vitro with one sperm, using micromanipulation. ICSI was done on an inverted Diaphot 300 microscope with Hoffmann modulation contrast and Narishige micromanipulators. Between March and May 1995 23 ICSI cycles were performed. Of 166 oocytes (7.2 per cycle) 22% were degenerated or immature and 6% damaged during ICSI. 45% of the injected eggs were fertilized. Embryos were transferred in 65% couples. Three pregnancies resulted (20%/ET), all are viable. ICSI is suitable in cases of severe oligospermia, asthenospermia and teratospermia, immunology infertility and low cleavage rate of oocytes in the first IVF cycle.

Adult↗

[Cryopreservation of embryos with the Cryogenetic apparatus].

For cryopreservation of human supernumerary embryos in our IVF programme we use a "Cryogenetic" computerised freezer. 1,2-propandiol and sucrose were used as cryoprotectant in the slow freezing/thawing protocol. In 20 transferred cycles we achieved 3 pregnancies. One is proceeds. Although the number of described cycles is low, the results are comparable with other centres. Our centre is the second one in the Czech Republic, which published data on a pregnancy after the transfer of cryopreserved embryos. Our Cryogenetic freezer is a high quality freezer suitable for gamete and embryo freezing.

Cryopreservation↗

[Evaluation of sperm morphology using the "strict" method].

The asset of evaluation of the morphology of sperm cells for prediction of their fertilizing power is generally accepted. Advances in the sphere of extracorporeal fertilization and the onset of micromanipulation techniques (intracytoplasmic injection of a single sperm cell - IC-SI) called for a stricter evaluation which is the prerequisite for a high standard indication and selection of different methods of extracorporeal fertilization. We obtained the consent of Prof. Gruger and Prof. Menkveld from the Tygerberg University (South African Republic) who elaborated this evaluation, to publish their criteria in our journal.

Humans↗

The human proteasome subunit HsN3 is located in the inner rings of the complex dimer.

Subunit HsN3 of the human proteasome is a beta-type subunit homologous to PRE4 from yeast, X1 beta from Xenopus and RN3 from the rat. Using electron microscopy, the binding sites of a monoclonal antibody with specificity for subunit HsN3 have been located in the two juxtaposed inner rings of the human proteasome. Subunit HsN3 was present in two copies, one in each ring, in accordance with our concept of two identical halves making up the complete human proteasome. The subunit is involved in the trypsin-like as well as the peptidylglutamyl-peptide cleavage activities.

Amino Acid Sequence↗

Characterization of the complete protein disulfide isomerase gene of Schistosoma mansoni and identification of the tissues of its expression.

cDNA and genomic clones of Schistosoma mansoni containing the complete sequence of a homolog of protein disulfide isomerase have been identified. The protein disulfide isomerase gene in schistosomes is a single copy gene having a genomic structure that is very similar to that of man. The C-terminus of the deduced protein is KDEL which in mammals functions as a signal sequence for retention of luminal proteins in the endoplasmic reticulum. Immunohistology and in situ hybridization identify the gastrodermis of the gut, the wall cells of the protonephridia, and the sustentacular cells of the testes to be the major tissues of protein disulfide isomerase gene expression. The protein disulfide isomerase of schistosomes, produced in an expression vector in Escherichia coli, catalyzes disulfide/sulfhydryl isomerization in vitro.

Amino Acid Sequence↗

In vitro activation of the 20S proteasome.

The effect of chemical compounds like sodium dodecyl sulfate (SDS), fatty acid esters of glycerol, carnitine and coenzyme A, phospholipids, histones, polylysines as well as homobifunctional chemical cross-linkers on the various proteolytic activities of mammalian proteasomes have been tested. Most of the reagents enhance these activities, and some, e.g. fatty acid CoA esters, histones and the chemical cross-linkers, exert dual effects, i.e. activation and inhibition at the same time, depending on the activity measured. With optimally activating concentrations of SDS, no structural changes in proteasomes can be detected by electron microscopy. Formation of micelles at supra-optimal detergent concentrations may be a reason for irreversible denaturation of the proteasome.

Amino Acid Sequence↗