J. Kawiak, A. Łukaszyk: Professor Piet van Duijn.
M. Gacko, A. Minarowska, A. Karwowska, Ł. Minarowski: Cathepsin D inhibitors.
A. Sulewska, W. Niklińska, M. Kozłowski, Ł. Minarowski, J. Nikliński, K. D±browska, L. Chyczewski: Detection of DNA methylation in eucaryotic cells. D. Bogolyubov, I. Stepanova: Interchromatin granule clusters in vitellogenic oocytes of the fleshfly, Sarcophaga sp.
Professor Piet van Duijn
(1921 - 2007)
Jerzy Kawiak and Andrzej Lukaszyk
Full text
The ImageStream System: a key step to a new era in imaging
Ewa K. Zuba-Surma1, Magdalena Kucia1, Ahmed Abdel-Latif3,
James W. Lillard Jr.2, and Mariusz Z. Ratajczak1,4
1Stem Cell Biology Program, University of Louisville, Louisville, KY, USA
2Brown Cancer Center, University of Louisville, Louisville, KY, USA
3Institute of Molecular Cardiology, University of Louisville, Louisville, KY, USA
4Department of Physiopathology, Pomeranian Medical University, Szczecin, Poland
Abstract: The aim of this article is to provide a brief review about the ImageStream system a novel tool for multiparameter
cell analysis in flow. The instrument integrates the features of flow cytometry and fluorescence microscopy combined
with a modern methodology for image analysis. Similar to flow cytometry, ImageStream allows analysis of a large number
of cells based on their fluorescence features and provides statistical analysis of these features. Additionally, ImageStream
allows detailed morphometric cellular analysis based on acquired cellular images integrating various morphometric and photometric
features of the examined cells. Simply stated, ImageStream system is an advanced flow cytometer acquiring both
integrated fluorescence signals as well as high quality fluorescence images and allowing muliparameter analysis. The innovative
features of the instrument offer new analytical capabilities and allow for a multitude of possible applications beyond
the current means of flow cytometry. While this article summarizes basic information about the features of ImageStream
and its applications based on the available literature and it also describes our own experience.
Marek Gacko1, Alina Minarowska2, Alicja Karwowska4, Łukasz Minarowski3
1Department of Vassal Surgery, Medical University of Bialystok, Poland
23rd Department of Children's Diseases, Medical University of Bialystok, Poland
3Department of Clinical Molecular Biology, Medical University of Bialystok, Poland
4Department of Instrumental Analysis, Medical University of Bialystok, Poland
Abstract: Inhibitors of cathepsin D belong to chemical compounds that estrify carboxyl groups of the Asp33 and Asp231
residues of its catalytic site, penta-peptides containing statin, i.e. the amino acid similar in structure to the tetraedric indirect
product, and polypeptides found in the spare organs of many plants and forming permanent noncovalent complexes with
cathepsin. Cathepsin D activity is also inhibited by alpha2-macroglobulin and antibodies directed against this enzyme.
Methods used to determine the activity and concentration of these inhibitors and their analytical, preparative and therapeutic
applications are discussed.
Detection of DNA methylation in eucaryotic cells
Anetta Sulewska1, Wieslawa Niklinska2, Miroslaw Kozlowski1, Lukasz Minarowski3 Wojciech Naumnik4, Jacek Niklinski1, Katarzyna Dabrowska3 and Lech Chyczewski3
Departments of: 1Thoracic Surgery, 2Histology and Embryology, 3Clinical Molecular Biology,
4Pneumonology and Tuberculosis, Medical University of Bialystok, Poland
Abstract: The methods of molecular biology allow for analyzing the methylation pattern in the whole genome and in particular
genes. We differentiate methylated sequences from unmethylated ones by means of cutting the genomic template with
methylation-sensitive restriction enzymes or by sodium bisulfite DNA modification. Chemical modification precedes most
quantitative and qualitative PCR techniques: MS-PCR, MS-nested PCR, Real-Time PCR, QAMA, HeavyMethyl, MSHRM.
Restriction enzymes, on the other hand, may be used together with PCR or hybridisation methods (Southern blot and
microarrays). PCRs are conducted with primers specific for methylated and unmethylated sequences and sometimes, similarly
to hybridisation techniques, with specifically labeled probes or dyes intercalating to double-stranded nucleic acids. The
most advanced methylation detection techniques (MALDI-TOF MS and HPLC) significantly reduce the amount of biological
material used for tests, but they require specialist equipment.
The localization of estrogen receptor ? and its function
in the ovaries of postmenopausal women
Agnieszka Brodowska1, Maria Laszczynska2, Andrzej Starczewski1,
Beata Karakiewicz3, Jacek Brodowski3
1Department of Reproduction and Gynaecology, 2Laboratory of Embryology, 3Laboratory of Family Nursing,
Pomeranian Medical University, Szczecin, Poland
Abstract: The localization of estrogen receptor ? (ER?) in the ovaries of postmenopausal women is a very up-to-date topic
in the aspect of using estrogens therapy in the clinical situations of different type. In ovaries of reproductive age women
ER? is present in ovary stroma, theca and granulosa cells, ovary surface epithelium (OSE) and in corpus luteum. The ovaries
of postmenopausal women are smaller than those of women at the reproductive age, the division into cortex and medulla
gets blurred, the ovaries have no follicles any longer, and the stroma is mainly composed of fibrous connective tissue, corpora
albicantia, nerves, and blood and lymphatic vessels. The aim of our study was to investigate the immunolocalization
and immunoexpression of ER? in the ovaries of postmenopausal women. The study involved 50 postmenopausal women
who had their ovaries removed by laparotomy due to non-neoplastic diseases of the uterus. The women were divided into 3
groups (A, B, and C) depending on the time that had passed since the last menstruation. Group A consisted of women who
had their last menstruation no more than 5 years earlier, in group B menopause occurred 5 to 10 years earlier, group C was
composed of patients who had the last menstruation over 10 years earlier. In all the patients concentrations of follicle stimulating
hormone (FSH), luteinizing stimulating hormone (LH), estradiol (E2), testosterone (T), androstendione (A) and
dehydroepiandrosterone sulphate (DHEAS) in blood plasma were measured. Ovarian tissue was obtained during surgery.
For morphological studies, ovaries were fixed in Bouin`s solution and 4% formalin and embedded in paraffin. Morphological
analysis was carried out after hematoxylin-eosin (HE) staining. Comparing to groups A and B, the ovaries in group C
contained a small number of corpora albicantia located in the medullary part as well as thinned blood vessels and few lymphatic
vessels and nerves. For immunoohistochemical expression of ER? paraffin-embedded specimens fixed in 4%
buffered formalin were used. The sections were next incubated with monoclonal mouse anti-human ER? antibody (N 1575
Dako, Denmark). Immunohistochemical nuclear expression of ER? in OSE, in epithelial inclusion cysts, in stroma, and in
group A also cytoplasmic expression of ER? in luteal and paraluteal cells of disappearing corpus luteum were revealed.
Immunohistochemical expression of ER? seems to decrease in the ovaries of women after menopause.
Prevalence of estrogen receptor ? PvuII and XbaI
polymorphism in population of Polish postmenopausal
women
Artur J. Jakimiuk1,2, Malgorzata Nowicka1, Michal Bogusiewicz3, Aneta Adamiak3,
Pawel Skorupski3, Pawel Miotla4, Tomasz Rechberger3, Jozef Haczynski3
1Department of Obstetrics and Gynecology, Central Clinical Hospital of Ministry of Interior
and Administration, Warsaw, Poland;
2Polish Academy of Sciences, Medical Research Center, Warsaw, Poland;
32nd Department of Gynecology, Medical University of Lublin, Poland;
4Chair and Department of Management and Health Protection Economics, Medical University of Lublin,
Poland
Abstract: Numerous data indicate that polymorphism of estrogen receptor ? (ER?) may predict lipid levels, lipid
response to hormone replacement therapy (HRT), myocardial infarction risk, bone fracture risk, bone mineral density
(BMD) and changes in BMD over time. In this study we aimed to evaluate distribution of ER? PvuII and XbaI genotypes
in population of Polish postmenopausal women qualified to different protocols of HRT. Subject of the study were 64 consecutive
postmenopausal women aged from 45 to 65 years (mean 56.6) assigned to HRT. ER? PvuII and XbaI polymorphism
was determined by PCR-restriction fragment length polymorphism (RFLP). The absence of PvuII and XbaI restriction
sites were indicated by "P" and "X" and presence by "p" and "x", respectively. PvuII genotype was distributed as follows:
PP 17.2% (n=11), Pp 50% (n=32), pp 32.83% (n=21). Frequency of XbaI genotype was: XX 6.25% (n=4), Xx
34.4% (n=22), xx 59.4% (n=38). Four haplotypes with following frequencies were recognized: PX 17.3%, px 47.4%, Px
24.4% and pX 10.9%. Prevalence of estrogen receptor ? PvuII and XbaI polymorphisms in Polish women is similar to
previously studied population.
Glucocorticoid receptor beta splice variant expression
in patients with high and low activity of systemic lupus
erythematosus
Piotr Piotrowski1,3, Michal Burzynski1, Margarita Lianeri1, Magdalena Mostowska1,
Mariusz Wudarski2, Hanna Chwalinska-Sadowska2, Pawel P. Jagodzinski1
1Department of Biochemistry and Molecular Biology, Poznan University of Medical Sciences, Poznan,
Poland;
2Institute of Rheumatology, Warsaw, Poland;
3Medical Research Center, Polish Academy of Sciences, Warsaw, Poland
Abstract: The glucocorticoid receptor (GR) occurs mainly in two alternative splice variants encoding GR? and GR?. The
GR? variant does not contain a GC binding domain and cannot mediate anti-inflammatory GC effects. Peripheral blood
mononuclear cells (PBMCs) were isolated from venous whole blood of twelve patients with SLE. Ten of the SLE patients
exhibited low disease activity while two patients displayed highly active stage of the disease. The quantitative analysis of
GR? and GR? transcripts in PBMC was performed by reverse transcription and real-time quantitative PCR SYBR Green I
system. The protein level of GR? and GR? isoforms in PBMCs was determined by western blotting analysis. We found that
the two SLE patients with high disease activity exhibited significantly elevated GR? transcript levels and corresponding protein
levels in PBMCs. These preliminary findings suggest that increased expression of GR? isoform may be associated with
relatively more severe clinical presentation of SLE syndrome.
Ex vivo measurement of calpain activation in human
peripheral blood lymphocytes by detection
of immunoreactive products of calpastatin degradation
Anna Mikosik1, Anna Zaremba1, Zofia Puchalska1, Agnieszka Daca1,
Zaneta Smolenska2, Paulina Lopatniuk1, Andrzej Mital3, Andrzej Hellmann3,
Ewa Bryl1 and Jacek M. Witkowski1
Departments of: 1Pathophysiology, 2Family Medicine, 3Hematology, Medical University of Gdansk,
Poland
Abstract: Limited proteolysis of multiple intracellular proteins by endogenous Ca-dependent cysteine proteases - calpains
- is an important regulatory mechanism for cell proliferation, apoptosis etc. Its importance for cellular functions is stressed
by existence of endogenous calpain inhibitors - calpastatins. The calpain-calpastatin system within living cells is in a fragile
balance, which depends on both partners. The interdependence of calpain - a protease - and calpastatin - an endogenous
inhibitor and at the same time a substrate for this enzyme makes any assessment of actual activity of this enzyme in the cells
very difficult. In this work we made an attempt to estimate and compare the activity of calpain in human peripheral blood
lymphocytes by assessing the levels of limited proteolysis of calpastatin in these cells by western blot, while at the same
time the levels of calpain protein inside these cells was measured by flow cytometry. Our results indicate that it is possible
to compare (semi-quantitatively) the activities of calpain in peripheral blood CD4+ and CD19+ lymphocytes from various
donors that way. Preliminary results showed that calpain activity is increased in the CD4+ T cells isolated from peripheral
blood of rheumatoid arthritis patients as compared to control lymphocytes. Extremely high intrinsic activity of calpain was
detected in chronic lymphocytic leukemia (CD19+) cells. All this confirms the detection of immunoreactive products of calpastatin
as a good maker of endogenous calpain activity.
Tissue localization of tumor antigen-loaded mouse
dendritic cells applied as an anti-tumor vaccine
and their influence on immune response
Joanna Rossowska, Elzbieta Pajtasz-Piasecka, Anna Szyda, Natalia Zietara,
Danuta Dus
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences,
Wroclaw, Poland
Abstract: The recognition, internalization and intracellular processing of antigen are the main functions of dendritic cells
(DCs). In the course of these processes, DCs differentiate and acquire the ability to produce cytokines responsible for polarization
of the immunological response. Therefore, vaccination with tumor antigen-loaded DCs is one of the most promising
approaches to induce tumor-specific immune response. The purpose of this study was to analyze the migratory abilities,
from an injection site to tumor-draining lymph nodes (tLN), of DCs applied as an anti-tumor vaccine and their capacity for
immune response activation. Mouse DCs of the established JAWS II cell line transduced with EGFP gene or ex vivo bone
marrow-isolated DCs (BM-DCs) stained with intravital CFDA dye were loaded with MC38 colon carcinoma tumor lysate
(TAg) and then administered peritumorally to MC38 tumor-bearing C57BL/6 mice. On the first, third, fifth and seventh days
after injection the tumors, tLNs and spleens were examined. The TAg-loaded DCs migrated more effectively to the tLNs
than did the unloaded control DCs; however, the majority of them remained in the tumor vicinity. Immunohistological analysis
of the tumor tissues demonstrated that only TAg-loaded DCs activated an immune response. Seven days after DCs vaccine
administration, numerous necrotic areas and some apoptotic bodies were observed in the tumor tissue. However, the
anti-MC38 tumor cytotoxic activity of spleen and tLN cells from mice treated with both TAg-loaded and unloaded DCs
reached a maximum on the fifth day after DC injection. Concluding, TAg-loaded DCs migrated more efficiently to tLNs and
were more effective activators of local (but not systemic) cellular immune response than were unloaded DCs. We hypothesize
that only the application of TAg-loaded DCs to tumor-bearing mice as an adjuvant supporting chemotherapy may activate
a more effective anti-tumor response.
Intracellular expression of the proliferative marker Ki-67
and viral proteins (NS3, NS5A and C) in chronic,
long lasting hepatitis C virus (HCV) infection
Aldona Kasprzak1, Agnieszka Adamek2, Wieslawa Biczysko3, Jolanta Seidel1,
Wieslawa Przybyszewska1, Karolina Olejniczak1, Jacek Juszczyk2, Maciej Zabel1,4
1Department of Histology and Embryology, University of Medical Sciences, Poznan;
2Department of Infectious Diseases, University of Medical Sciences, Poznan;
3Department of Clinical Pathomorphology, University of Medical Sciences, Poznan;
4Department of Histology and Embryology, University of Medical Sciences, Wroclaw, Poland
Abstract: Hepatitis C virus (HCV) continues to represent the main causative agent of the hepatitis, which leads to chronic
transformation of the process in 60-80% individuals. It remains unclear how far cellular expression of HCV proteins in vivo
may represent an index of progression of the disease and of proliferative activity in the liver in chronic hepatitis C. Aim of
the studies included detection and subcellular localization of three HCV proteins (NS3, NS5A and C) in liver biopsies from
adults (n=19) with chronic, long lasting hepatitis C as related to hepatocyte proliferative activity. The immunocytochemical
ABC (avidin biotin-peroxidase complex) technique was applied, alone or associated with the ImmunoMax technique.
Results of the immunocytochemical tests were compared to histological alterations in liver biopsies, proliferation index and
with selected clinical data. Asignificantly higher expression of NS3 protein was noted, as compared to expressions of NS5A
and C proteins. In all the patients, cytoplasmic localization of all proteins dominated over nuclear localization (p<0.05). At
the level of electron microscopy, protein localization in endoplasmic reticulum (ER) membranes, mitochondria, perinuclear
region and/or in hepatocyte cell nucleus was observed. No direct relationships could be demonstrated between expressions
of HCV proteins and of Ki-67 antigen. No correlations could also be demonstrated between cellular expression of any HCV
protein on one hand and grading or staging, alanine transaminase (ALT), serum level of HCV RNA or alpha-fetoprotein
(AFP) on the other. However, positive correlations were disclosed between proliferative activity of hepatocytes on one hand
and patient's age, grading and staging on the other. Advanced hepatic fibrosis correlated also with serum levels of AFP. The
studies were supplemented with data on subcellular localization of HCV proteins. Moreover, they indicated that in HCV
infection grading and staging, proliferative activity of hepatocytes and serum AFP level represent more valuable indices of
the disease progress than those provided by cellular expression of three potentially oncogenic HCV proteins in vivo.
Cytochemical and immunocytochemical studies
of the localization of histones and protamine-type
proteins in spermatids of Chara vulgaris
and Chara tomentosa
Katarzyna Poplonska, Agnieszka Wojtczak, Maria Kwiatkowska,
Andrzej Kazmierczak
Department of Cytophysiology, University of Lodz, Poland
Abstract: Spermiogenesis in Chara algae, which has been divided into 10 phases (sp I-X), is similar to spermiogenesis in
animals. The most important process during spermiogenesis in animals is remodeling of chromatin leading to "sleeping
genome", being the result the exchange of histone proteins into protamine-like proteins. Cytochemical studies showed in
both Chara species (C. vulgaris, C. tomentosa) that at spI-IV phases only histones were present, at spV-VIII phases - the
amount of nuclear protamine-type proteins progressively increased and that of histones decreased while at spIX-X only protamine-
type proteins were present. This was also confirmed with capillar electrophoresis. In order to localize more precisely
both histones and protamines the immunocytochemical studies with the use of anti-protamine antibodies (protamine-type
proteins were obtained from C. tomentosa antheridia) and anti-histone H3 antibodies, have been carried out. More specific
immunocytochemical studies confirmed cytochemical results including the exchange of histones into protamine-type during
spermiogenesis (spV-VIII) in both Chara species. At phase V spermiogenesis these strong strand-like anti-protamine
signals were observed in cytoplasm which might suggest that protamine synthesis took place in ER.
Expression and cellular distribution of NADPH-diaphorase
and nitric oxide synthases in the porcine
uterus during early pregnancy
Aneta Andronowska, Marcin Chrusciel
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Olsztyn, Poland
Abstract: Nitric oxide plays a key role in the regulation of various female reproductive processes such as ovulation, implantation
and myometrial relaxation. The aim of the present study was to determine the histochemical activity and cellular localization
of NADPH-d in the porcine uterus during early pregnancy, including the implantation period. Tissue samples collected
from the pig uteri on days 5, 10, 12, 15 and 17 of pregnancy were stained histochemically for NADPH-d activity and
immunohistochemically for NOS isoforms localization. In the luminal epithelium a significant increase of NADPH-d activity
was observed on days 5 - 12 of pregnancy. On day 17 of pregnancy, two different staining patterns were observed: 1) a
significant (p<0.001) decrease in NADPH-d activity at the site of implantation and 2) the high NADPH-d activity at interimplantation
regions. The endometrial glands showed a significant (p<0.001) increase in NADPH-d staining with high activity
in individual glands. The arterial endothelium expressed stronger NADPH-d staining compared with venous vessels.
Immunoreactivity of eNOS was similar to NADPH-d staining but no optical differences in the intensity of staining were
observed. Clear iNOS immunoreactivity was detected in the luminal epithelium, endometrial stroma and individual endometrial
glands. The vascular endothelium displayed weak iNOS staining.
11?-hydroxysteroid dehydrogenase type 2 expression
in the newly formed Leydig cells after ethane
dimethanesulphonate treatment of adult rats
Yvetta Koeva1, Mariana Bakalska2, Nina Atanassova2, Katerina Georgieva3
and Michail Davidoff4
1Department of Anatomy and Histology, Medical University, Plovdiv, Bulgaria;
2Institute of Experimental Morphology and Anthropology with Museum, Bulgarian Academy of Sciences,
Sofia, Bulgaria;
3Department of Physiology, Medical University, Plovdiv, Bulgaria;
4Institute of Anatomy 1, UKE, University of Hamburg, Germany
Abstract: The enzyme 11?-hydroxysteroid dehydrogenase (11?-HSD) catalyzes the reversible conversion of physiologically
active corticosterone to the biologically inert 11?-dehydrocorticosterone in rat testis and protect the Leydig cells (LCs)
against the suppressive effect of glucocorticoids. The developmental pathway of the adult LCs population is accompanied
with an increase in the 11?-HDS activity. Thus, 11?-HDS together with its role in controlling the toxicological effect of glucocorticoids
on LCs can be used as a marker for their functional maturity. Ethane 1,2-dimethanesulphonate (EDS) treatment
of adult rats become unique appropriate model, which enable to answer many questions related to the differentiation of adult
LCs in the prepubertal rat testis. The aim of the present study was to investigate the specific changes in the 11?-HDS type
2 immunoreactivity in tandem with the expression of androgen receptor (AR) during renewal of LCs population after EDS
treatment. In the present study, we observed the first appearance of immunostaining for 11?-HSD2 in new LCs population
on day 14 after EDS administration when the progenitor LCs were detected. Our immunohistochemical analysis revealed
progressive increases in the 11?-HSD2 reaction intensity on 21 days after EDS treatment and reached a maximum on day
35. AR immunoexpression was found in new LCs on day 14 and 21 after EDS injection with an increasing curve of intensity.
The most prominent AR immunostaining in new population LCs was evident by 35 days after EDS and that coincided
with the increased number of LCs and restoration of adult LCs population. Our results demonstrated similar pattern of
immunoreactivity for 11?-HSD2 and AR in new LCs population after EDS treatment and suggested that the changes in 11?-
HSD2 expression can be used for evaluation of adult LCs differentiation in rat testis.
Interleukin-6 is not essential for bone turnover in hypothyroid mice
Janusz Mysliwiec1, Robert Zbucki2, Maria M. Winnicka2, Boguslaw Sawicki3,
Agnieszka Nikolajuk1, Karol Kaminski4, Piotr Mysliwiec5, Wlodzimierz Musial4,
Zofia Bondyra6, Jerzy Walecki6, Maria Gorska1
Departments of: 1Endocrinology, Diabetology and Internal Diseases, 2General and Experimental
Pathology, 3Histology and Embryology, 4Cardiology, 5II Department of General Surgery, 6Radiology,
Medical University of Bialystok, Bialystok, Poland
Abstract: Abstract: Interleukin-6 (IL-6) has been shown to be involved in the pathogenesis of several bone diseases characterized by
an imbalance between bone resorption and formation. The aim of the study was to estimate serum markers of bone turnover:
osteoclast-derived tartrate-resistant acid phosphatase form 5a (TRACP 5b) and osteocalcin in IL-6-deficient mice to assess
the role of IL-6 in bone metabolism in hypothyroidism in mice. C57BL/6J (wild-type; WT) and C57BL/6JIL6-/-Kopf (IL-6
knock-out; IL6KO) mice randomly divided into 4 groups with 10 in each one: 1/ WT mice in hypothyroidism (WT-ht), 2/
WT controls, 3/ IL6KO mice with hypothyroidism (IL6KO-ht) and 4/ IL6KO controls. Experimental model of hypothyroidism
was induced by intraperitoneal injection of propylthiouracyl. The serum levels of TRACP 5b and osteocalcin were
determined by ELISA. Serum concentrations of TRACP 5b (median and interquartile ranges) were significantly decreased
in both groups of mice with hypothyroidism: WT (3.2 (2.5-4.7) U/l) and IL6KO (2.6 (1.8-3.5) U/l) as compared to the
respective controls. Similarly, serum osteocalcin levels were significantly reduced in both groups of mice in experimental
hypothyroidism: WT (25.8 (23.0-28.2) ng/ml) and IL6KO (21.5(19.0-24.6) ng/ml) in comparison to the respective controls.
There were no significant differences in bone turnover markers between IL6KO and WT mice both in hypothyroid and control
animals. The results of the present study suggest that IL-6 does not play an important role in bone turnover in both euthyroid
and hypothyroid mice.
Effect of soil and water environment on typeability
of PowerPlex Y (Promega) in selected tissue samples
Anna Niemcunowicz-Janica1, Witold Pepinski1, Jacek Robert Janica2,
Malgorzata Skawronska1, Jerzy Janica1, Ewa Koc-Zorawska1, Ireneusz Stolyszewski3
1Department of Forensic Medicine, Medical University of Bialystok, Poland
2Department of Radiology, Medical University of Bialystok, Poland
3Department of Criminalistics and Forensic Medicine, University of Warmia and Mazury, Olsztyn, Poland
Abstract: In cases of decomposed bodies Y chromosomal STR markers may be useful in identification of a male relative.
The authors assessed typeability PowerPlex Y (Promega) loci in tissue material stored in water and soil environment. Tissue
material was collected during autopsies of five persons aged 20-30 years with time of death determined within the limit
of 14 hours. Heart muscle, liver and lung specimens were stored in pond water, sea water, sand and peat soil. DNA was
extracted by organic method from tissue samples collected in 7-day intervals. Liver specimens were typeable in all PowerPlex
Y loci within 100 days of storage in pond water with gradual decline at DYS392 in sea water. Heart muscle specimens
stored in pond water exhibited allelic loss at DYS19, DYS385, DYS389II and DYS392, while all loci were typeable
in sea water stored samples. For lung specimens allelic loss was noted throughout the profile. Storage of liver specimens in
peat soil for more than 14 days resulted in allelic drop-out, and after 21 days no profiles were typeable. Heart muscle specimens
were typeable in all PowerPlex Y systems after 35-day storage in sand, while allelic drop-out and subsequent lack of
profiles were noted after 14 and 35 days respectively. Lung specimens stored in garden soil exhibited allelic drop-out and
subsequent lack of profiles after 7 and 21 days, respectively. All PowerPlex Y loci were typeable in the latter material in
sand up to day 35 with gradual decline of longer amplicons (DYS19, DYS385, DYS389II and DYS392)
Changes of mitochondrial pathway in hypoxia/reoxygenation induced cardiomyocytes apoptosis
Aina He1, Jian-an Wang1, Chun Gui2, Yun Jiang1, Yong Sun1, Tielong Chen1
1Heart Center, Second Affiliated Hospital, College of Medicine, Zhejiang University, 88 Jiefang
Road,Hangzhou, 310009, China;
2Heart Center, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, 3 East Qingchun
Road, Hangzhou, 310016, China
Abstract: The role of mitochondrial apoptotic pathway in cardiomyocytes subjected to hypoxia/reoxygenation(H/R) was
studied. Cultured cardiomyocytes from neonatal Sprague-Dawley rats were exposed to hyoxia/reoxygenation, the apoptotic
cardiomyocytes were stained with Annexin-V-FITC, Hoechst 33342 and TUNEL assay. Mitochondrial transmembrane
potential of cardiomyocytes was assessed by JC-1 under fluorescence microscope, the expressions of bcl-2, bax, cytochrome
c, apoptosis-induced factor (AIF), and caspase-3 were tested by western-blot. Our data showed apoptosis of cardiomyocytes
was significantly increased during H/R, accompanied by translocation of bax to mitochondria, release of cytochrome c and
AIF to cytosol. The results indicate that the mitochondrial-mediated apoptotic pathway is initiated as a result of H/R.
Interchromatin granule clusters in vitellogenic oocytes of the fleshfly, Sarcophaga sp.
Dmitry Bogolyubov, Irina Stepanova
Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia
Abstract: Insect oocyte nuclei contain different extrachromosomal nuclear bodies including Cajal bodies and interchromatin
granule clusters (IGCs). In the present study, we describe IGC equivalents in the vitellogenic oocytes of the fleshfly,
Sarcophaga sp. These structures were found to consist of 20-40-nm granules and also include the fibrillar areas of high and
low electron density. Immunogold labeling electron microscopy revealed IGC marker protein SC35, Sm proteins, and
trimethylguanosine cap of small nuclear (sn) RNAs in these bodies. Antibody against the non-phosphorylated RNA polymerase
II selectively labeled the fibrillar areas of low electron density located inside the IGCs.
The presence of blaIMP genes on plasmids DNA isolated
from multidrug - resistant Pseudomonas aeruginosa
strains at University Hospital in Bialystok (Poland) -
first report
Pawel Sacha1, Marcin Zorawski2, Tomasz Hauschild3, Piotr Wieczorek1,
Jadwiga Jaworowska4, Piotr Jakoniuk1, Elzbieta Tryniszewska1,4
1Department of Microbiological Diagnostics, Medical University of Bialystok, Poland
2Department of Pharmacology, Medical University of Bialystok, Poland
3Department of Microbiology, Institute of Biology, University of Bialystok, Poland
4Department of Microbiological Diagnostics, University Hospital of Bialystok, Poland
Abstract: Resistance to carbapenems is emerging, and it is a great problem to therapeutics. Seven multidrug-resistant
(MDR) of Pseudomonas aeruginosa strains were isolated from urine and bronchial specimens. All isolates showed resistance
to imipenem and meropenem (MIC; >16 mg/L). The resistance to carbapenems in two of seven strains was associated
with the production of a metallo-beta-lactamases. Plasmids DNA probes were used to investigate the presence of genes coding
for IMP-type enzymes. PCR experiments revealed that blaIMP genes were present in two isolates of Pseudomonas aeruginosa
(MIC >32 ug/mL for both carbapenems).