Volume 10 (2005) pp 3-13 |
Title |
THE EFFECT OF AGE ON THE MORPHOMETRIC SPERM TRAITS
OF DOMESTIC PIGS (Sus scrofa domestica) |
Authors |
Stanisław Kondracki*, Dorota Banaszewska
and Cecylia Mielnicka |
Abstract |
The experiment was conducted using 20 male domestic pigs, which
were in 2 (equal-sized) age groups: under 14 months old and over 18 months
old. At least 5 ejaculates from each male were taken, and in each ejaculate,
morphometric measurements of 50 spermatozoa were made. The measured
parameters were head area, head length and width, and flagellum length. For
each ejaculate, the basic physical traits and the frequency of occurrence of
developmental anomalies of the spermatozoa were examined. It was found that
sperm dimensions tended to increase along with the boars' age. Considerable and
statistical differences in head area and flagellum length were proved.
Spermatozoa collected from older boars (above 18 months of age) had a head
area larger by 0.49 µm2 (P≤0.01) and a flagellum longer by 0.67 μm (P≤0.01)
than spermatozoa collected from younger boars (under 14 months of age). The
differences in head length and width between the spermatozoa of the tested
boars were considerably smaller and were not statistically provable. Significant
correlations between the head area of spermatozoa and the head length (r = 0.56,
P≤0.05) and head width (r = 0.36, P≤0.05) were found. However, the head
length was not significantly directly correlated with its width (r = 0.18, P≤0.05),
and flagellum length was negatively correlated with spermatozoan head width (r
= -0.71, P≤0.05). Slight correlations between the morphometric traits of the
sperm and the physical traits of the ejaculates (r = -0.11 to 0.16) were found,
although in most cases, the correlations were not statistically provable. |
Address and Contact Information |
Department of Bioengineering and Animal Breeding, University of Podlasie,
B. Prusa 14, 08-110 Siedlce, Poland *Corresponding author; e-mail: sk@ap.siedlce.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 15-21 |
Title |
APOPTOSIS AND CYTOTOXICITY CAUSED BY ETHOXYQUIN AND
TWO OF ITS SALTS |
Authors |
Alina Błaszczyk1* and Janusz Skolimowski2 |
Abstract |
In our study, we analyzed the cytotoxicity of ethoxyquin (EQ) and its
two salts, ethoxyquin hydrochloride (EQ-HCL) and ethoxyquin phosphate (EQP).
It was shown that EQ was the most cytotoxic compound (IC50 = 0.09 mM),
while the lowest cytotoxic effect was observed for EQ-P (IC50 = 0.8 mM). The
properties of ethoxyquin and its salts were also analyzed with the TUNEL
method, which evaluates their ability to induce apoptosis. It was shown that EQ
induced apoptosis in cultured human lymphocytes, especially at concentrations
of 0.25 and 0.5 mM. |
Address and Contact Information |
1Department of Cytogenetics and Plant Molecular Biology, University of Łódź,
Banacha 12/16, 90-237 Łódź, Poland, 2Department of Organic Chemistry,
University of Łódź, Narutowicza 68, 90-136 Łódź, Poland *Corresponding author; e-mail: ablasz@biol.uni.lodz.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 23-35 |
Title |
THE LOADING OF HUMAN ERYTHROCYTES WITH SMALL
MOLECULES BY ELECTROPORATION |
Authors |
Gintautas Saulis* |
Abstract |
The technology for loading the cell with membrane-impermeable
substances by means of electroporation consists of the following three stages: (i)
the creation of pores permeable for the desired substance; (ii) the introduction of
a substance into the cell cytosol; and (iii) the restoration of the membrane barrier
function.
In this paper, the experimental data on the loading of human erythrocytes with
small molecules (molecular weight < 500 Da) is presented. The results obtained
show that increasing the intensity of the electric field pulse increases the fraction
of electroporated cells. The pores through which the molecules of ascorbic acid
and mannitol (radius < 0.5 nm) can enter the erythrocytes appear when the field
strength exceeds 2.5 kV/cm. The concentration of ascorbic acid inside the cells
increases linearly. At 4oC, the rate of ascorbic acid influx was constant for at
least 4 hours. The original permeability of most of the cells towards ascorbic
acid and mannitol was restored after about 6-7 min at 37oC, and the
characteristic time for complete resealing was about 20-40 min. The procedure
described here can be used for loading cells with membrane-impermeable
substances. |
Address and Contact Information |
Biophysical Research Group, Department of Biology, Faculty of Nature Science,
Vytautas Magnus University, 28 Daukanto str., Kaunas 44246, Lithuania tel: +370612-82773, fax: +370-37-406572, e-mail: sg@kaunas.omnitel.net |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 37-47 |
Title |
REPEATED INJECTIONS OF PEG-PE LIPOSOMES GENERATE
ANTI-PEG ANTIBODIES |
Authors |
Kamila Środa1,3*, Janusz Rydlewski2, Marek Langner1,
Arkadiusz Kozubek3, Michał Grzybek3 and Aleksander F.
Sikorski1,3** |
Abstract |
Liposomes containing the polyethylene glycol (PEG) derivative of
phosphatidyl ethanolamine (PE) have recently been found to be promising drug
carriers, as they facilitate controlled and target-oriented release of therapeutics.
They also reduce the side effects of many drugs. Here, we present the results of a
study on antiliposomal properties of rabbit sera obtained after weekly injections
of small liposomes containing 20% PEG-PE. The effect was analysed as the
level of induced carboxyfluorescein release from these liposomes in vitro. The
incubation of liposomes with rabbit serum taken after the injections induced the
release of carboxyfluorescein at a higher level than was seen for incubation with
untreated animal’s serum. The strongest effect was observed for serum obtained
after the second injection, i.e. during the second week of the study. The effect
was much smaller after the serum samples were preheated at 56oC. The binding of serum proteins by PEGylated liposomes was analysed via gel filtration and
via the immunoblot technique using goat anti-rabbit IgG; this revealed that the
serum protein which bound to the liposomes in vitro had a molecular weight of
~55 kD and reacted with the anti-IgG antibody. Competition with PEG or lipids
indicate that this IgG has an anti-PEG activity. We therefore assume that these
antibodies are responsible for the activation of complement and leakage
induction of PEG-liposomes. Such antibodies could be responsible for increased
phagocytosis by RES macrophages (in particular liver macrophages) and
decreased circulation time. |
Address and Contact Information |
1Academic Centre for the Biotechnology of Lipid Aggregates,
Przybyszewskiego 63/77, 51-148 Wrocław, Poland, 2Department and Clinic of
Internal Veterinary Medicine, Agricultural University of Wrocław, pl.
Grunwaldzki 47, Wrocław, Poland, 3Institute of Biochemistry and Molecular
Biology, University of Wrocław, Przybyszewskiego 63/77, 51-148 Wrocław,
Poland, 4Institute of Physics, Wrocław University of Technology, Wybrzeże
Wyspiałskiego 27, 50-370 Wrocław, Poland *Present address: Department of Biophysics, Wrocław Medical University,
Chałubiłskiego 10, 50-368 Wrocław, Poland
**Corresponding author; tel: +4871 375 6233, fax: +4871 375 6208, e-mail:
afsbc@ibmb.uni.wroc.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 49-59 |
Title |
DISTRIBUTION OF TIGHT DNA-PROTEIN COMPLEXES ALONG THE
BARLEY CHROMOSOME 1H, AS REVEALED BY MICROSATELLITE
MARKER ANALYSIS. |
Authors |
Tatjana Sjakste1,2, Marion Röder1, Danute Labeikytė3, Lida
Bagdoniene3, Anna Levina4, Benediktas Juodka3
and Nikolajs Sjakste4 |
Abstract |
The distribution of DNA complexes with proteins resistant to routine
deproteinisation procedures (tightly bound proteins, TBP) was studied on the
barley chromosome 1H by means of microsatellite analysis. The polypeptide
spectrum of the barley shoot TBP was similar to that formerly described for other
organisms. In order to reveal developmental changes in the distribution of the
TBP, DNA was extracted from dry grains, coleoptiles, root tips, and young and
old leaves. In the seeds, all the studied DNA sites were evenly distributed
between free DNA and DNA containing the tight DNA-protein complexes.
Germination made the interaction between TBP and chromosomal loci specific.
In coleoptile DNA, sites containing microsatellites located in the distal part of the
long arm of the chromosome were not bound to the TBP anymore, however, the
centromeric markers were found exclusively in the tight DNA-protein
complexes. A similar but not identical distribution of markers was observed in
the root tips and young leaves. Leaf senescence was accompanied by a loss in
interaction specificity between chromosomal loci and tightly bound proteins.
These results are considered to reflect changes in chromatin domain interaction
with the nuclear matrix during plant development. |
Address and Contact Information |
1Institute for Plant Genetics and Crop Research, Correnstrasse 3, Gatersleben,
Germany, 2Laboratory of Plant Genetics, Institute of Biology, University of
Latvia, Miera iela 3, Salaspils LV2169, Latvia, 3Department of Biochemistry and
Biophysics, Vilnius University, M. K. Čiurlionio 21, LT-2009 Vilnius, Lithuania,
4Faculty of Medicine, University of Latvia, Šarlotes iela 1a, Riga LV-1001, Latvia |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 61-72 |
Title |
SPECTROSCOPIC STUDIES ON THE PROTECTIVE EFFECT OF A
SPECIFIC SUGAR ON CONCANAVALIN A AT ACIDIC, NEUTRAL
AND ALKALINE PH. |
Authors |
Rizwan Hasan Khan1*, Aabgeena Naeem1 and Masroor Alam
Baig2 |
Abstract |
A Systematic investigation of the effect of pH on concanavalin A in
the presence of specific and non-specific sugars is made using CD (circular
dichroism) and fluorescence. The specific and non-specific sugars for
concanavalin A were methyl α-D-glucopyranoside and methyl α-Dgalactopyranoside
respectively. Far-UV CD showed changes in the MRE value
at 217 nm in the presence of the above-mentioned sugars. At pH 7, the CD and
fluorescence spectra obtained in the presence of methyl α-D-glucopyranoside
were slightly different from those for the native state and a significant difference
was obtained in the presence of methyl α-D-galactopyranoside. Near-UV CD
spectra showed the retention of a native-like tertiary structure in the presence of
the specific sugar upon pH denaturation. Tryptophan fluorescence studies
indicated a change in the tryptophan enviornment. The results obtained from our
CD data are consistent with those obtained from fluorescence studies.
Upon pH exposure of concanavalin A in the presence of methyl α-Dglucopyranoside
and methyl α-D-galactopyranoside, the former acted as a
protector preventing conformational alteration at different pH while the presence
of latter induced a different stable conformational state and this state persists
over the pH range from 2 to 10. |
Address and Contact Information |
1Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh-
202002, India, 2Jamia Hamdard University, Hamdard Nagar, New Delhi
110 062, India *Corresponding author; fax: 91-571-2701766, tel: 91-571-27020388, E-mail:
rizwanhkhan@hotmail.com, rizwanhkhan1@yahoo.com |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp73-89 |
Title |
A SUPPORT VECTOR MACHINE APPROACH TO THE
IDENTIFICATION OF PHOSPHORYLATION SITES. |
Authors |
Dariusz Plewczyński1,2*, Adrian Tkacz3, Adam Godzik4,5
and Leszek Rychlewski1 |
Abstract |
We describe a bioinformatics tool that can be used to predict the
position of phosphorylation sites in proteins based only on sequence
information. The method uses the support vector machine (SVM) statistical
learning theory. The statistical models for phosphorylation by various types of
kinases are built using a dataset of short (9-amino acid long) sequence
fragments. The sequence segments are dissected around post-translationally
modified sites of proteins that are on the current release of the Swiss-Prot
database, and that were experimentally confirmed to be phosphorylated by any
kinase. We represent them as vectors in a multidimensional abstract space of
short sequence fragments. The prediction method is as follows. First, a given
query protein sequence is dissected into overlapping short segments. All the
fragments are then projected into the multidimensional space of sequence
fragments via a collection of different representations. Those points are
classified with pre-built statistical models (the SVM method with linear,
polynomial and radial kernel functions) either as phosphorylated or inactive
ones. The resulting list of plausible sites for phosphorylation by various types of
kinases in the query protein is returned to the user. The efficiency of the method
for each type of phosphorylation is estimated using leave-one-out tests and
presented here. The sensitivities of the models can reach over 70%, depending
on the type of kinase. The additional information from profile representations of
short sequence fragments helps in gaining a higher degree of accuracy in some phosphorylation types. The further development of an automatic
phosphorylation site annotation predictor based on our algorithm should yield a
significant improvement when using statistical algorithms in order to quantify
the results. |
Address and Contact Information |
1BioInfoBank Institute, ul. Limanowskiego 24A/16, 60-744 Poznań, Poland,
2Interdisciplinary Centre for Mathematical and Computational Modeling,
University of Warsaw, ul. Pawiłskiego 5a, 02-106 Warsaw, Poland,
3Bioinformatics Unit, Department of Physics, Adam Mickiewicz University, ul.
Umultowska 85, 61-614 Poznań, Poland, 4The Burnham Institute, La Jolla, CA,
USA, 5Bioinformatics Core JCSG, University of California San Diego, La Jolla,
CA, USA *Corresponding author; tel: +48-61-8653520, fax: +48-61-8643350, e-mail:
darman@bioinfo.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 91-99 |
Title |
THE SHAPE OF CELLS ADHERING TO SULFONATED COPOLYMER
SURFACES. |
Authors |
Hanna M. Kowalczyńska*, Małgorzata Nowak-Wyrzykowska,
Marcin Inkielman, Liliana Stołowska
and Ewa Marciniak |
Abstract |
We studied the shape of L1210 leukaemia cells adhering in a proteinfree
medium to sulfonated (styrene/methyl methacrylate) copolymer surfaces of
two sulfonic group densities, and thus of differing wettability. The use of our
image analysis method and the mathematical procedure [1] allowed us to
calculate the values of the so-called shape parameter, which quantitatively
determines the three-dimensional cell shape. Here, we show that the values of
the shape parameter of the adhering cells and the F-actin concentration, in the
region near the cell-substratum interface, depend on the density of sulfonic
groups present on the substratum surface.. |
Address and Contact Information |
Department of Biophysics and Biomathematics, Medical Centre for Postgraduate
Education, ul. Marymoncka 99, 01-813 Warszawa, Poland *Corresponding author; fax: +48 22 8640834, e-mail: hmkowal@cmkp.edu.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 101-121 |
Title |
THE MYSTERY OF PHOSPHOLIPID FLIP-FLOP IN BIOGENIC
MEMBRANES. |
Authors |
Sathyanarayana N. Gummadi* and Krishna S. Kumar |
Abstract |
Phospholipid flip-flop is required for bilayer assembly and the
maintenance of biogenic (self-synthesizing) membranes such as the eukaryotic
endoplasmic reticulum and the bacterial cytoplasmic membrane. Due to the
membrane topology of phospholipid biosynthesis, newly synthesized
phospholipids are initially located in the cytoplasmic leaflet of biogenic
membranes and must be translocated to the exoplasmic leaflet to give uniform
bilayer growth. It is clear from many studies that phospholipid flip-flop in
biogenic membranes occurs very rapidly, within a period of a few minutes.
These studies also reveal that phospholipid translocation in biogenic membranes
occurs bi-directionally, independently of the phospholipid head group, via a
facilitated diffusion process in the absence of metabolic energy input, and that
this type of transport requires specific membrane proteins. These translocators
have been termed biogenic membrane flippases, and they differ from metabolic
energy-dependent transporters (ABC transporters and MDR proteins). No
biogenic membrane flippases have been characterized. This review briefly
discusses the importance of biogenic membrane flippases, the various assay
methods used for measuring the rate of phospholipid flip-flop, and the progress
that has been made towards identifying these proteins. |
Address and Contact Information |
Department of Biotechnology, Indian Institute of Technology-Madras, Chennai
600 036, India *Corresponding author, e-mail: gummadi@iitm.ac.in |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 123-134 |
Title |
A PCR-BASED MOLECULAR MARKER APPLICABLE FOR
MARKER-ASSISTED SELECTION FOR ANTHRACNOSE DISEASE
RESISTANCE IN LUPIN BREEDING. |
Authors |
Mingpei You1, Jeffrey G. Boersma2, Bevan J. Buirchell1,2,
Mark W. Sweetingham1,2, Kadambot H. M. Siddique1
and Huaan Yang1,2* |
Abstract |
Selection for anthracnose disease resistance is one of the major
objectives in lupin breeding programs. The aim of this study was to develop
a molecular marker linked to a gene conferring anthracnose resistance in narrowleafed
lupin (Lupinus angustifolius L.), which can be widely used for MAS in
lupin breeding. A F8 derived RIL population from a cross between cultivar
Tanjil (resistant to anthracnose) and Unicrop (susceptible) was used for marker
development. DNA fingerprinting was conducted on 12 representative plants by
combining the AFLP method with primers designed based on conserved
sequences of plant disease resistance genes. A co-dominant candidate marker
was detected from a DNA fingerprint. The candidate marker was cloned,
sequenced, and converted into a sequence-specific, simple PCR based marker.
Linkage analysis based on a segregating population consisting of 184 RILs
suggested that the marker, designated as AntjM2, is located 2.3 cM away from
the R gene conferring anthracnose resistance in L. angustifolius. The marker has
now being implemented for MAS in the Australian national lupin breeding
program. |
Address and Contact Information |
1Centre for Legumes in Mediterranean Agriculture (CLIMA), Faculty of Natural
and Agricultural Sciences, The University of Western Australia, Crawley, WA
6009, Australia, 2Department of Agriculture Western Australia, 3Baron-Hay
Court, South Perth, WA 6151, Australia * Corresponding author, e-mail: hyang@agric.wa.gov.au
|
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 135-149 |
Title |
EXPRESSION OF HUMAN MEMBRANE SKELETON PROTEIN
GENES FOR PROTEIN 4.1 AND bIIÎŁ2-SPECTRIN ASSAYED BY
REAL-TIME RT-PCR # |
Authors |
Pamela M. Taylor-Harris1*, Leanne E. Felkin2*,
Emma J. Birks2, Rodney C.G. Franklin3, Magdi H. Yacoub2,
Anthony J. Baines4, Paul J.R. Barton2 and Jennifer C. Pinder1** |
Abstract |
The proteins, spectrin and 4.1 confer support and resilience to animal
cell membranes, and promote assembly of multimeric, membrane-bound
signalling complexes. Protein 4.1 also plays important roles in tumour
suppression and the regulation of cell proliferation. To assess relative tissue
expression of the four genes encoding human protein 4.1, we measured mRNA
levels using quantitative real-time polymerase chain reaction. We compared 4.1
expression with that of a major splice variant of spectrin, bIIS2 that has
a shortened C-terminus lacking a pleckstrin homology domain. mRNA for 4.1R
is four-fold higher in bone marrow than in tissues with the next highest
prevalence: cerebellum, lung, testis and thymus. 4.1G mRNA is highly
expressed in brain, spinal cord and testis; 4.1N in brain, spinal cord and adrenal
gland; 4.1B in testis, brain, spinal cord, and kidney. Thus, 4.1N, 4.1B and 4.1G
all show high accumulation in nervous tissues. mRNA for bIIÎŁ2-spectrin is
ubiquitous, but most abundant in cardiac and nervous tissues. Comparative transcript abundance was analysed in heart and brain. bIIS2-spectrin was the
most abundant transcript in heart with levels 5 fold greater than 4.1G or 4.1N
and at least 9 fold greater than 4.1B. In brain, 4.1N was the most abundant
transcript, with levels 2.4 fold greater than 4.1B and at least 4 fold greater than
4.1G or bIIS2-spectrin. 4.1R abundance was very low in both tissues. Whilst we
expected that 4.1 mRNAs would feature highly in muscle and nerve, we note
their high abundance in testis, indicating previously unsuspected functions in
reproduction. |
Address and Contact Information |
1Randall Division of Cell and Molecular Biophysics, King's College London,
Guy's Campus, London SE1 1UL, UK, 2Heart Science Centre, Imperial College
London, Harefield, Middlesex UB9 6JH, UK,3Royal Brompton & Harefield
NHS Trust, Harefield, Middlesex UB9 6JH, UK 4Department of Biosciences,
University of Kent, Canterbury, Kent, CT2 7NJ, UK # Contribution to the Conference “Emerging Pathways in Cytoskeletal Communication”,
Umea, Sweden 2004, organised by CytoNET
* These authors made equal contributions to the work
**Corresponding author; fax: +44-20-7848-6435, e-mail: jennifer.fordham@kcl.ac.uk |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 151-161 |
Title |
RAPD ANALYSIS OF THE INTERSPECIFIC SOMATIC HYBRIDS
Solanum bulbocastanum (+) S. tuberosum |
Authors |
Danuta Bołtowicz*, Anna Szczerbakowa
and Bernard Wielgat |
Abstract |
The diploid Mexican species S. bulbocastanum (blb) was used as a
source of late blight resistance in somatic hybridization with the potato (S.
tuberosum, tbr) dihaploid H-8105. The produced 2x blb (+) 2x tbr H-8105
somatic hybrids did not retain the blb parent's characteristic high resistance to P.
infestans. The revealed aneuploidy of blb (+) tbr H-8105 hybrids indicated a
possible loss of individual blb chromosome(s) carrying the resistance genes.
Four hybrid clones differing in terms of their ploidy, morphology and growth
potential were analysed for the presence of all twelve blb chromosomes (linkage
groups). The RAPD markers assigned to particular chromosomes were selected
on the basis of the linkage map of S. bulbocastanum constructed by Naess et al.,
Mol. Gen. Genom. 265 (2001) 694-704. Of the 86 markers analysed, twelve
(14%) were common for blb and H-8105, while 34 (40%) and 40 (46%) markers
were specific for the blb and H-8105 genome, respectively; this confirms the
differences between the nuclear genomes of the two species. Seventeen markers
(20%) present in one or the other parent were absent in the hybrids, and only one
new marker was found in the hybrids. The poorly growing, aneuploid and
chimeric hybrids had the same band profiles as the well growing,
morphologically normal hybrids, except for two bands that were present only in
normal hybrids. The presence of eleven blb linkage groups in the blb (+) tbr H-
8105 hybrids was confirmed. The markers specific for the second linkage group
(chromosome 2) of blb were not present in the RAPD patterns of the somatic
hybrids analysed, suggesting a loss or rearrangement of this chromosome in the
combined nuclear genome of the hybrids. |
Address and Contact Information |
Institute of Biochemistry and Biophysics, Polish Academy of Sciences,
Pawiłskiego 5A, 02-106 Warsaw, Poland *Corresponding author, e-mail: danabo@ibb.waw.pl |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 163-171 |
Title |
A CAPS MARKER SET FOR MAPPING IN LINKAGE GROUP III OF
PEA (Pisum sativum L.) |
Authors |
Fedor Konovalov, Eugenia Toshchakova
and Sergei Gostimsky |
Abstract |
A set of twelve CAPS markers was mapped for linkage group III of
pea (Pisum sativum L.). New primers were designed to use a polymerase chain
reaction to amplify fragments of sequenced pea genes containing at least one
large intron. Amplification products were tested for polymorphism across three
pea lines (Chi115, Flagman and WL1238) using eleven four-base restriction
endonucleases. Nine STS markers for linkage group III from the literature were
also tested for polymorphism, and five of these were used in this mapping study
as anchor points. All polymorphic loci were located by genetic analysis of the F2
population from the cross Chi115 x WL1238, and a map of linkage group III
consisting of one morphological and twelve CAPS markers was created. The
map covers the full length of the chromosome and is about 162 cM long. All the
CAPS markers in a set were used to test for polymorphism among 10 additional
pea DNA samples extracted from different marker lines and cultivars. |
Address and Contact Information |
Genetics Department, Moscow State University, Vorobjovy Gory, Moscow,
119899, Russia *Corresponding author, e-mail: konovalov@pisumsativum.org |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 173-183 |
Title |
STABILITY AND AGGREGATION STUDIES OF NON-SONICATED
ARSONOLIPID-CONTAINING VESICLES |
Authors |
Dimitrios G. Fatouros1,*, Nikolaos Bouropoulos2,
Panayiotis V. Ioannou3 and Sophia G. Antimisiaris1 |
Abstract |
In this study, we investigated non-sonicated arsonolipid-containing
liposomes (arsonoliposomes) in terms of the influence of lipid composition on
their stability, assessed as membrane integrity and physical stability [size].
Vesicles consisting of plain arsonolipids or mixtures of arsonolipids with
cholesterol [Chol] or with distearoyl-phospatidylcholine [DSPC] were studied.
Membrane integrity was evaluated by measuring the retention of incorporated
5-(6)carboxyfluorescein [CF] during incubation of the vesicles in Tris buffer, pH
7.4. Photon correlation spectroscopy was used to investigate the time-dependent
aggregation of arsonoliposomes in the absence and presence of Ca2+ ions.
Vesicles composed of plain C18 (acyl fatty chain) arsonolipids were found to be
unstable, with only 15% of the initially incorporated CF remaining in the
vesicles after 24 hours. The addition of Chol to the membrane (1:1 mol/mol)
significantly increased the stability of arsonoliposomes, while the addition of
DSPC to the lipid bilayer (1:1 mol/mol) increased vesicle stability to a lower
extent. The results of particle size analysis showed that non-sonicated
arsonoliposomes consisting of plain arsonolipid Ars/Stearic are highly and
rapidly aggregated, while calcium-induced aggregation is also significant, but
slower. Aggregation could not be always explained on the basis of zeta potential
changes, indicating that the process is complex. |
Address and Contact Information |
1Laboratory of Pharmaceutical Technology, Department of Pharmacy,
University of Patras, 26504, Greece, 2Department of Materials Science,
University of Patras, 26504, Greece, 3Department of Chemistry, University of
Patras, Patras, 26504, Greece *Corresponding author; current address: Department of Pharmaceutics, The Danish
University of Pharmaceutical Sciences, Universitetsparken 2, 2100, Copenhagen,
Denmark, tel: +45 35306288, fax: +45 35306030, e-mail:df@dfuni.dk |
![[Rozmiar: 1312 bajtółw]](pic/pdf.gif) ![[Rozmiar: 1332 bajtółw]](pic/abstract.gif) |
Volume 10 (2005) pp 185-193 |
Title |
MOLECULAR CLONING AND CHARACTERIZATION OF A NOVEL
HUMAN GENE CONTAINING 4 ANKYRIN REPEAT DOMAINS |
Authors |
Jixi Li, Chaoneng Ji, Huarui Zheng, Xiangwei Fei, Mei Zheng,
Jianfeng Dai, Shaohua Gu, Yi Xie and Yumin Mao* |
Abstract |
Ankyrin repeat, one of the most important protein motifs, plays a
wide variety of roles in protein-protein interactions and in the signal pathways.
Via large-scale sequencing, a novel 941-bp gene was isolated from an 18-week
old human fetal brain cDNA library. It encodes a putative protein of 158 amino
acid residues with four conserved ankyrin repeat domains. It displays a high
degree of homology with rat low-density lipoprotein receptor-related protein 2-
binding protein (Lrp2bp), and was therefore was named hLrp2bp (human
Lrp2bp). The hLrp2bp gene was located in chromosome 4q35 and the conserved
ankyrin repeat domains were located between amino acid residues 10 and 116.
RT-PCR revealed that hLrp2bp was mainly expressed in the human testis, small
intestine, colon and blood leukocytes, and in human pancreatic adenocarcinoma
cells. A HEK293 cell was transfected with the ORF of hLrp2bp, and analyses
showed that the protein was distributed both in the cytoplasm and nucleus. |
Address and Contact Information |
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of
Life Sciences, Fudan University, Shanghai 200433, People's Republic of China *Corresponding author; tel: +86-21-65643573, fax: +86-21-65642502, e-mail:
ymmao@fudan.edu.cn |
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