Contribution
of stem cells to skeletal muscle regeneration
2Department of Cytology, Faculty of
Biology, Warsaw University;
3Department of Experimental
Hematology, Maria Skłodowska-Curie Memorial Cancer Center and Institute
of
Oncology, Warsaw, Poland
Abstract: Stem cells
for skeletal muscle originate from
dermomyotome of the embryo. The early marker of these cells is
expression of
both transcription factors Pax3 and Pax7 (Pax3+/Pax7+
cells). The skeletal muscles in the adult organism have a remarkable
ability to
regenerate. Skeletal muscle damage induces degenerative phase, followed
by
activation of inflammatory and satellite cells. The satellite
cells are quiescent myogenic
precursor cells located between the basal
membrane and the sarcolemma of myofiber and they are characterized by Pax7
expression. Activation
of the satellite cells is regulated by muscle growth and chemokines.
Apart from
the satellite cells, a population of adult stem cells (muscle side
population - mSP) exists in the skeletal muscles. Moreover, the cells
trafficking from different tissues may be involved in the regeneration
of
damaged muscle. Trafficking of cells in the process of damaged muscle
regeneration may be traced in the SCID mice.
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text
1Institute of Experimental Morphology
and Anthropology, Bulgarian Academy of Sciences, Sofia,
Bulgaria;
2Department of Anatomy and Histology,
Higher Medical Institute, Plovdiv, Bulgaria;
3Institute of Anatomy 1, UKE,
University of Hamburg, Germany
Abstract: Androgens are especially important
for the maintenance of spermatogenesis in adulthood and the
experimental
withdrawal of testosterone (T) production by ethane dimenthanesulfonate
(EDS)
is a valuable tool for studying androgen-dependent events of
spermatogenesis.
The aim of the present study was to investigate the specific changes in
immunoexpression of androgen receptor (AR) in the testis in relation to
degeneration and regeneration of Leydig cell (LC) population and
seminiferous
epithelium. Immunohistochemistry for AR and 3β-hydroxysteroid dehydrogenase (3β-HSD) as well as TUNEL assay for
apoptosis were performed on testicular sections of control and
EDS-treated rats.
Serum LH and T levels were measured by RIA. Our results revealed a
total loss
of AR immunoexpression from the nuclei of Sertoli (SCs), LCs and
peritubular
cells during the first week after EDS administration and that coincided
with
severe drop in T levels. Two weeks after EDS administration, the AR
expression
was recovered in these cells but normal stage-specificity in SCs was
replaced
by uniform intensity of AR immunostaining at all the stages of the
spermatogenic cycle. The stage-specific pattern of androgen expression
in SCs
with a maximum at stages VII-VIII appeared 5 weeks after treatment. LC
immunoreactivity for 3β-HSD
at different time points after EDS administration correlated with
values of T
concentration. The maximal germ cell apoptosis on day 7 was followed by
total
loss of elongated spermatids 2 weeks after EDS treatment. Regeneration
of
seminiferous epithelium 3 weeks after EDS administration and onwards
occurred
in tandem with the development of new LC population indicated by the
appearance
of 3β-HSD-positive
cells and gradual increase in T production. The specific changes in AR
after
EDS including their loss and recovery in Sertoli cells paralleled with
degenerative and regenerative events in Leydig and germ cell
populations,
confirming close functional relationship between Sertoli, Leydig and
germ
cells.
Author’s
e-mail: ninaatanassova@yahoo.com
N.
Negić, N. Nestorović, M. Manojlović-Stojanoski, B. Filipović, B.
©oąić-Jurjević, V. Milosević and M. Sekulić
Institute
for Biological Research Siniąa Stanković, Belgrade, Serbia and
Montenegro
Abstract:
Exposure to
glucocorticoids leads to numerous changes in various biological systems
including the reproductive system. The aim of the present work was to
find out
whether dexamethasone (Dx) treatment of adult female rats would
influence the
histological and morphometric characteristics of the pituitary
gonadotrophic
cells (luteinizing - LH cells and follicle stimulating - FSH cells).
One group
of female Wistar rats received Dx injections on three consecutive days
in doses
1.0, 0.5 and 0.5 mg/kg b.w. respectively, while the control rats were
treated
with equivalent volumes of saline. Experimental and control animals
were
sacrificed 24 h and 72 h after the last injection. The
peroxidase-antiperoxidase (PAP) immunocytochemical procedure was used
to study
the LH and FSH cells. The stereological and morphometric analyses
showed that
multiple Dx treatments of female rats significantly decreased the
volume of LH
cells and the volume of their nuclei 24 h and 72 h after the last Dx
injection
in comparison with control values. At 24 h after Dx treatment, the
volume
density of LH cells was significantly increased, but at 72 h
differences
between the experimental and control groups were insignificant. The
increase in
number of LH cells per unit area (mm2) was significant at
both
timepoints (24 h and 72 h). Stereologic and morphometric
characteristics of FSH
cells was changed after Dx treatment in the same manner
as that of LH cells, except for the volume density, where a
significant
increase was established 24 h and 72 h after the last Dx application.
These
results clearly demonstrate that 24 h and 72 h after the last of three
Dx
injections there were changes in the immunocytochemical and
morphometric
features of gonadotrophic cells.
Author’s
e-mail:
negicn@ibiss.bg.ac.yu
NK
cell
depletion and recovery in SCID mice treated with anti-NK1.1 antibody
Joanna Kulesza, Grażyna Hoser, Danuta
Wasilewska and
Jerzy Kawiak
Department
of Clinical Cytology, Medical Center of Postgraduate Education,
Warszawa,
Poland
Abstract: The anti-NK1.1 antibody produced by
PK136 hybridoma cell line administered subcutaneously to SCID mice
effectively
decreased the level of peripheral blood NK cells and weight of the
spleen for
3-4 days. The antibody treatment did not harm the general state of the
animal,
and may be practically applied in xenograft experiments.
Author’s
e-mail:
jkawiak@cmkp.edu.pl
Mobilization
of human hematopoietic stem/progenitor-enriched CD34+ cells
into peripheral blood during stress related to ischemic stroke
2Clinic of Neurology, Pomeranian
Medical University, Szczecin;
3Department of Transplantology,
Children’s Hospital, Jagiellonian
University Medical College, Kraków, Poland
Abstract:
The
bone marrow-derived stem/progenitor cells were demonstrated to play an
important role in a regeneration of damaged tissue. Based on these
observations
we asked whether the stroke-related stress triggers mobilization of
stem/progenitor cells from the bone marrow into the peripheral blood,
which
subsequently could contribute to regeneration of damaged organs. To
address
this issue, the peripheral blood samples were harvested from patients
with
ischemic stroke during the first 24 hrs as well as after the 48 (2nd
day) and
144 hrs (6th day) since the manifestation of symptoms. In these
patients we
evaluated the percentage of hematopoietic stem/progenitor-enriched CD34+
cells by employing flow cytometry and the number of hematopoietic
progenitor
cells for the granulocyto-monocytic (CFU-GM) and erythroid
(BFU-E)-lineages
circulating in peripheral blood. We concluded that stress related to
ischemic
stroke triggers the mobilization of hematopoietic stem/progenitor cells
from
the bone marrow into peripheral blood. These circulating
stem/progenitor cells
may play an important role in the process of regeneration of the
ischemic
tissue.
Interleukin-2
(IL-2) expression in livers of patients with chronic hepatitis C virus
(HCV)
infection
Aldona Kasprzak1, Jolanta Seidel1,
Agnieszka Adamek2, Wiesława Biczysko3, Jacek
Wysocki4,
Rafał Spachacz1, Jacek Juszczyk5 and Maciej Zabel1,6
2Ward of Infectious Diseases, J.
Stru¶ Hospital, Poznań
3Department of Clinical
Pathomorphology, University of Medical Sciences, Poznań;
4Department of Health Prophylaxis,
University of Medical Sciences, Poznań;
5Department of Infectious Diseases,
University of Medical Sciences, Poznań;
6Department of Histology and
Embryology, University of Medical Sciences, Wrocław, Poland
Author’s e-mail: akasprza@amp.edu.pl
Stromal
myofibroblasts in breast cancer: relations between their occurrence,
tumour
grade and expression of some tumor markers
Paweł
Surowiak1,2,3, Sławomir Suchocki4, Balazs Györffy2,5,
Tserenchunt Gansukh2,6, Andrzej Wojnar3, Adam
Maciejczyk3,
Marek Pudełko3 and Maciej Zabel1,7
2Institute of Pathology, Charité
Campus Mitte, Berlin, Germany;
3Lower Silesian Centre of Oncology, Wrocław, Poland;
4Department of Obsterics and
Gynaecology, Voivodship Hospital, Walbrzych, Poland;
5Joint Laboratory of the Hungarian
Academy of Sciences and the Semmelweis University, 2nd Department of Internal Medicine, Semmelweis
University, Budapest, Hungary;
6Health Sciences University of
Mongolia, Ulaanbaatar, Mongolia;
7Department of Histology and
Embryology, University School of Medicine, Poznań, Poland
Abstract: It is suggested that tumour stromal
myofibroblasts exert an unfavourable effect on the biology of breast
cancer. We
are aware of only a single study which examined relationships between
manifestation of myofibroblasts in the stroma of breast cancer and
clinicopathological data of the patients. The present study was aimed
at
estimation of the effect exerted by myofibroblasts present in the
tumour stroma
on principal pathological parameters and on expression of Ki67, P53 and
HER-2
proteins in the group of the most frequent breast cancers, the ductal
cancers.
In paraffin sections of 60 ductal breast cancers (20 cases in G1, 20 in
G2 and
20 in G3), immunohistochemical reactions were performed to detect
expression of
smooth muscle actin (SMA) in order to visualize myofibroblasts, Ki67,
P53 and
HER-2. The studies demonstrated that the most numerous myofibroblasts
were
present in G3 cases and they were the least frequent in G1 cases (P=0.02).
Positive correlations were observed between the presence of
myofibroblasts in
tumour stroma and expression of Ki67 and HER-2 in breast cancer cells
in the
entire group (P<0.001 and P=0.001, respectively), in
G2 cases
(P=0.003 and P=0.03) and in G3 cases (P=0.01 and P=0.03).
Considering that the higher grade, Ki67 and HER-2 are thought to
represent
unfavourable prognostic factors, the elevated content of myofibroblasts
in
tumour stroma is probably typical for cases with worse prognosis.
Author’s
e-mail:
pawel.surowiak@interia.pl
Evaluation
of sperm genomic integrity of normozoospermic men: a prospective study
M. Piasecka1, D. G±czarzewicz2
and
M. Laszczyńska3
1Departament of Histology and
Embryology, Pomeranian Medical University; 2Department of
Animal
Reproduction, University of Agriculture; 3Laboratory of
Embryology,
Pomeranian Medical University, Szczecin, Poland
Abstract: The objective of our study was to
evaluate the incidence of spermatozoa with nuclear DNA strand breaks in
patients with normal routine sperm parameters (26 subjects). Sperm DNA
fragmentation was measured using TUNEL test assessed in flow cytometer.
Variable percentages of sperm with damaged DNA (9.42±7.68%; range:
2-36) were
found. Two categories of patients were distinguished: (1) patients (8
out of 26
subjects) with ≤4% of TUNEL-positive sperm and (2) patients (18 out of
26
subjects) with >4% of TUNEL-positive sperm. A significantly lower
percentage
of normal sperm forms was found in patients with >4% of
TUNEL-positive sperm
than in patients with ≤4% of TUNEL-positive sperm. Moreover, a
significant
negative correlation (rs=-0.50) was noted only between a
proportion
of normal sperm forms and a proportion of TUNEL-positive spermatozoa.
In
electron microscope, a large number of spermatozoa with immature
chromatin was
observed more frequently in subjects with >4% of TUNEL-positive
cells (11
out of 18 subjects). Our results suggest that in some patients with
normal
routine sperm parameters, DNA fragmentation may be associated with poor
sperm
morphology. The diminished sperm genomic integrity may result from
molecular
disturbances in nuclear remodeling process during spermiogenesis. TUNEL
assay
is a screening tool that may help to discriminate between fertile and
infertile
men and may help to predict successful in vitro fertilization.
Localization
of the DAZ gene expression in seminiferous tubules of patients
with
spermatogenic disorders
Anna Szczerba, Anna Jankowska, Mirosław
Andrusiewicz and
Jerzy B. Warchoł
Department
of Cell Biology, University of Medical Sciences, Poznań, Poland
Abstract: The research on the expression and
mutations of DAZ and its homologues in human and other mammals
suggests
that protein products of these genes can mainly affect development of
germinal
cells. The aim of the present study was to analyze the expression of
the DAZ
gene in seminiferous tubules of six men with spermatogenic disorders
(hypospermatogenesis and spermatogenic arrest). The results based on the RT-PCR IS technique
demonstrated that the DAZ product was present only in some
seminiferous
tubules and the fluorescence intensity was different within individual
tubules.
The most intense fluorescence characterised the spermatogonia,
especially these
organised in small groups inside separate tubules. In the patients with
spermatogenic arrest at the spermatocyte stage, the DAZ gene
transcripts
were also found in primary spermatocytes. However, the fluorescence
intensity
of primary spermatocytes, except the fluorescence of the spermatocytes
localised upon the lumen, was weaker than the fluorescence of
spermatogonia.
The results of our study showed that DAZ gene activity seems to
correspond to the proliferative activity of stem cells of germinal
epithelium.
DNA
damage induced by mutagens in plant and human cell nuclei in acellular
comet
assay
Jolanta Juchimiuk, Agnieszka Gnys and Jolanta
Maluszynska
Department
of Plant Anatomy and Cytology, University of Silesia, Katowice, Poland
Abstract: Higher plant cells have a long
tradition of use in the studies on environmental mutagenesis in situ,
especially in relation to human health risk determination. The studies
on the
response of plant and human cells to physical and chemical mutagens
showed differences
in their sensitivity. The differences in the presence of cell
components in
plants and humans could influence such response. Additionally, the
level of the
organization of the employed material could influence DNA-damaging
effect:
leukocytes are isolated cells and plant – an intact organism. To
preclude these
obstacles, the effects of direct treatment of isolated nuclei with
genotoxic
agents were determined to compare the sensitivity of plant and human
cells. In the
present study, we have determined the DNA-damaging effects of two
chemical
mutagens: maleic acid hydrazide (MH) and N-methyl-N-nitroso-urea (MNU)
applied
to isolated nuclei of both plant and human cells. In order to compare
the
sensitivity of the nuclei of Nicotiana tabacum var. xanthi and
the nuclei of leukocytes, the acellular Comet assay was carried out.
The
results showed higher sensitivity of the nuclei of leukocytes as
compared to
the nuclei of plant cells to mutagenic treatment with the applied doses
of MH
and MNU.
Microtubules
with different diameter, protofilament number and protofilament spacing
in Ornithogalum
umbellatum ovary epidermis cells
Maria Kwiatkowska1, Katarzyna
Popłońska1,
Dariusz Stępiński1 and Zygmunt Hejnowicz2
1Department of Cytophysiology,
University of ŁódĽ;
2Department of Biophysics and Biology
of Cell, Silesian University, Katowice, Poland
Abstract: Microtubules present in the
epidermis of Ornithogalum umbellatum ovary in the area of
lipotubuloids
(i.e. aggregates of lipid bodies surrounded by microtubules) are 25–51
nm in
diameter. They consist mainly of 10 and 11, sometimes 9 and 12
protofilaments.
An average diameter of microtubule consisting of 9 subunits is about 32
nm, of
10 – 35 nm, of 11 – 38 nm and of 12 – 43 nm, however, individual
microtubules
in each category significantly vary in size. These differences result
from
varying distance between protofilaments in microtubule walls and
diameters of
protofilaments: in thin microtubules they are densely packed and
smaller while
in thicker ones they are loosely arranged and bigger.