bluuebunny/crossref_metadata_2025 huggingface dataset:
Enrichment of blood from embryonic stem cells in vitro
Authors: Perkins Andrew C.
Published: November 2003

Murine embryonic stem (ES) cells are pluripotent. When injected into
blastocysts they can give rise to every cell type of a derived chimeric mouse
including germ cells. Embryonic stem cells also possess remarkable
in vitro differentiation potential. When removed from
stromal support and leukaemia inhibitory factor (LIF), ES cells differentiate
into structures known as embryoid bodies (EBs), in which all three germ layers
develop and interact. As ES cells from humans become available there is
increasing interest in the potential for EBs to provide an unlimited supply of
stem cells for somatic transplantation therapies. Realisation of this
potential will require greater understanding of the molecular determinants of
cell fate within EBs. Also, culture techniques for selective expansion of cell
lineages of interest will reduce the risks associated with transplantation of
EB-derived cells. In this paper the kinetics of expression of mRNA and protein
for early mesoderm markers within EBs is reported. In addition, a three-step
culture system incorporating co-cultivation on the bone marrow derived stromal
cell line, MC3T3-G2/PA6 (PA6), is explored as a way to select for
haematopoietic progenitor cells (HPCs) and against undifferentiated ES cells.
A system like this could enhance purification of haematopoietic stem cells
(HSCs) from ES cells for bone marrow transplantation.
Activity of platelet-activating-factor-acetylhydrolase and the nitric oxide metabolite level in the plasma of pregnant women who develop transient hypertension during later pregnancy
Authors: Okumura Kyoko K. , Sagawa Norimasa , Kobayashi Fuminori , Nanno Hidetaka , Matsumoto Tsunekazu , Itoh Hiroaki , Korita Daizo , Tanada Shozo , Mori Takahide
Published: November 2003

It is reported that plasma platelet-activating-factor-acetylhydrolase (PAF-AH)
is elevated in patients with essential hypertension. In this study, plasma
PAF-AH activity was measured during pregnancy and after delivery to examine
the relationship between plasma PAF-AH activity and the development of
transient hypertension (TH) during pregnancy. Moreover, in order to examine
the involvement of endothelial injury in TH, the plasma level of nitric oxide
metabolite (NOx; NO2+NO3) was
measured. The plasma PAF-AH activity in 51 pregnant women was consecutively
measured in the 1st, 2nd and 3rd trimesters of gestation, and after delivery.
Forty-one cases were normal pregnancies and 10 cases were complicated by TH
later during pregnancy. The PAF-AH activity in the normal pregnancy group
decreased in the 2nd trimester of gestation compared with the 1st trimester,
but was elevated in the TH group. The incidence of elevation of PAF-AH in the
TH group was significantly (7/10; 70.0%;
P<0.01, Chi-squared test) higher than in the normal
pregnancy group (9/41; 22.0%). The plasma NOx levels in the 2nd
trimester were higher than those in the 1st trimester in both the normotensive
and TH group (P<0.05 for both comparisons). The 51
patients were classified into two groups according to the change in the PAF-AH
in the 2nd trimester: group A consisted of 35 patients whose PAF-AH activity
did not increase, and group B consisted of 16 patients whose PAF-AH activity
increased. The incidence of development of TH during later pregnancy in group
B was significantly (7/16; 43.8%; P<0.01,
Chi-squared test) higher than in group A (3/35; 8.6%). Hypertension
developed after 36 weeks’ gestation in all patients in the TH group. The
results of the present study suggest that changes in PAF metabolism may relate
to regulation of blood pressure in pregnant women whose pregnancy is
complicated with TH, whereas NO metabolism does not differ between women with
TH and those having a normal pregnancy.
Extra keyword: PAF-acetylhydrolase
Immunoelectronmicroscopic imaging of spermadhesin AWN epitopes on boar spermatozoa bound in vivo to the zona pellucida
Authors: Rodríguez-Martínez H. , Iborra A. , Martínez P. , Calvete J. J.
Published: November 2003

Spermadhesin AWN is a major protein of boar seminal plasma and a sperm
surface-associated lectin. AWN binds to β-galactosides and to porcine
zona pellucida glycoproteins, suggesting a role for this protein in primary
gamete interaction. However, because capacitation induces remodelling of the
sperm surface and AWN is peripherally bound to the plasma membrane, the
present study sought to investigate whether AWN is present or absent in the
subpopulation of spermatozoa that reaches the ovulated oocyte at the period of
fertilization in vivo. Therefore, tubal tissues and
oocytes from sows mated with a fertile boar were collected 6–8 h after
ovulation. Tissues and oocyte–sperm complexes were fixed, immunolabelled
with anti-AWN monoclonal antibodies, and examined by means of light and
scanning electron microscopy. The results show that spermadhesin AWN is
present in spermatozoa seen along the genital tract of the natural mated sow
as well as on plasmalemmal remnants of spermatozoa bound to the zona pellucida
in vivo.
Seasonal variation in ovarian response to pregnant mares serum gonadotrophin in the brushtail possum (Trichosurus vulpecula)
Authors: Glazier A. M.
Published: November 2003

This study examined the seasonal variation in ovarian response to the
exogenous hormone pregnant mares serum gonadotrophin (PMSG) in the brushtail
possum (Trichosurus vulpecula). Ovarian stimulation was
achieved by administration of a single intramuscular injection of 15 IU PMSG.
Animals (n = 165) responded to this treatment in
a variable manner depending on the month of the year. The maximum response
(measured by the total number of large (>2 mm) follicles) was 15.8 ±
1.5 follicles per animal (n = 12) in May and the
minimum response was 4.9 ± 1.6 follicles per animal
(n = 7) in January. The response throughout the
rest of the year closely paralleled the birth seasonal distribution observed
in wild possums in published reports. In the summer months (December, January
and February) when few females carried pouch young, 55%, 42% and
17% of PMSG-primed animals, respectively, exhibited a nil response to
PMSG (i.e. no follicles >2 mm were found on the ovaries of treated
animals). In addition, when wild possums were in lactational anoestrus after
the autumnal breeding peak, possums with a nil response to PMSG accounted for
17% (July) and 6% (August) of those treated, but the other
animals gave near maximal numbers of large follicles (July, 5–21;
August, 2–25). Whether the ovaries of non-responders are downregulated
in some manner outside the breeding season or they become insensitive to
gonadotrophin remains to be investigated. These observations pinpoint times of
the year when the highest productivity can be expected following PMSG
treatment, and together with information on the timing of ovulation provide
critical data for the development of artificial breeding strategies for the
possum.
Minimum sperm trajectory length for reliable determination of the fractal dimension
Authors: Mortimer Sharon T.
Published: November 2003

The aim of this study was to determine the minimum track length required for
the reliable calculation of the fractal dimension of trajectories of
capacitating human spermatozoa. Manually reconstructed trajectories classified
previously as hyperactivated or non-hyperactivated were re-analyzed. The
trajectories were reconstructed at 60 Hz, and each comprised 61 points
(corresponding to 1 s movement). The trajectories were ‘split’ to
give the equivalent of 2 × 31 point, 3 × 21 point, 4 × 16
point and 6 × 11 point track segments and the fractal dimension
determined for each. The fractal dimensions of each track segment within each
trajectory were compared using paired t-tests. No
significant difference was observed between the fractal dimensions of track
segments of equal length, irrespective of the motility pattern. However,
significant differences in fractal dimension values were observed when
segments of different lengths were compared (P<0.01).
For the non-hyperactivated tracks, the fractal dimension was consistently
below the hyperactivation threshold level for only the 31 point segments. The
hyperactivated tracks consistently had fractal dimension values above the
threshold level when segments of 16 points or longer were analysed. Therefore,
the minimum track length for the determination of reliable fractal dimension
values was 31 points, corresponding to an image sampling time of 0.5 s at 60
Hz, although it could be used as a screening method for the identification of
hyperactivated motility for track segments of greater than 11 points
(corresponding to 0.17 s movement).
Effects of pre- and post-mating nutritional status on hepatic function, progesterone concentration, uterine protein secretion and embryo survival in Meishan pigs
Authors: Ashworth C. J. , Beattie L. , Antipatis C.
Published: November 2003

This experiment examined whether the pre- or the post-mating diet had greater
impact on embryo survival in Meishan gilts. Gilts received either a
maintenance (1.15 kg day −1 ;
n=12) or a high (3.5 kg day
−1 ; n=12) diet
during the oestrous cycle preceding mating. After mating, half the animals in
each group received either the maintenance or the high diet until slaughter on
Day 12. Gilts fed the high pre-mating diet had more corpora lutea (22.7
v.19.0, SED = 0.98;
P<0.001), increased embryo survival (95.5%
v.74.8%, SED=7.58;
P<0.01) and heavier corpora lutea (−0.71 log g
v.−0.90 log g, SED=0.09;
P=0.07) compared with gilts fed the maintenance
diet prior to mating. The post-mating diet had no effect on embryo survival.
There were no treatment effects on blastocyst developmental stage, luteal
surface area or progesterone release. Gilts receiving the high post-mating
diet had heavier livers than those fed the maintenance post-mating diet (1.45
v.1.08% of total bodyweight, SED=0.07;
P<0.001), suggesting that these gilts have a greater
capacity to metabolize progesterone. Pre-mating nutritional status therefore
appears to be a greater determinant of embryo numbers and survival than the
post-mating diet.
The use of DNA fingerprinting to assess monozygotic twinning in Meishan and Landrace × Large White pigs
Authors: Ashworth C. J. , Ross A. W. , Barrett P.
Published: November 2003

The extent of embryo mortality is usually estimated by the difference between
the numbers of corpora lutea and embryos in the same animal, which assumes
that each corpora lutea represents one potential embryo. Recent observations
from the authors' laboratory reveal situations in which the number of
embryos exceeds the number of corpora lutea, implicating the presence of
identical twins. The objective of the present study was to establish DNA
fingerprinting techniques to investigate the prevalence of monozygotic
twinning in two breeds of pig. DNA fingerprints of every fetus carried by 6
Meishan (MS) and 6 Landrace x Large White (LxLW) gilts on Day 29
± 2 of pregnancy were obtained. Five LxLW and 3 MS litters
carried no identical fetuses. The remaining gilts carried a pair of fetuses
with indistinguishable DNA profiles. No pairs of fetuses were monochorionic.
These results suggest that monozygotic twinning in the pig occurs during
embryo cleavage or blastocyst development and may be more prevalent in MS
compared with indigenous breeds.
Carboxylesterase overexpression in the male reproductive tract: a universal safeguarding mechanism?
Authors: Mikhailov A. T. , Torrado M.
Published: November 2003

Data on expression patterns of carboxylesterases in the male reproductive
tract of different animal groups (i.e. bivalve mollusks, fruitflies and
rodents) are summarized to highlight some particularly interesting questions
in the context of sperm differentiation, maturation and function. The male
reproduc-tive system, in spite of extreme variation in the
anatomical/morphological organization in different species, is
characterized by similar patterns of male-dependent carboxylesterase
overexpression. The phenomenon of conserved carboxylesterase overexpression
indicates similar male sex-associated functions of the enzymes. There is
possible evidence of carboxylesterase recruitment by male reproductive-tract
tissues indi-cating that it could be adaptive for spermatogenesis, sperm
maturation and sperm use. Moreover, this idea can be extended to include a
sperm cell lineage protection. This issue is discussed in the light of recent
data on environmental reproductive xenobiotics that can provide a basis for a
hypothetical explanation of car-boxylesterase overexpression in the male
reproductive tract. Based on a well-known role of car-boxylesterases in
detoxification of environmental chemicals such as organophosphate pesticides,
it is proposed that various male genital tract carboxylesterases may be
characterized by a similar physiological function to protect the male
reproductive system against xenobiotic influences that could provoke its
dys-function, thus altering sperm differentiation and maturation.
Effects of unilateral ovariectomy with compensatory hypertrophy on ovarian blood flow, oxygen consumption and progestin secretion in the 16-day pregnant rat
Authors: Bruce N. W. , Massa H. M.
Published: November 2003

Stimulated ovulation with resultant multiple corpora lutea (CL) can result in
lower proges-terone levels than expected from the increased luteal tissue
mass. Unilateral ovariectomy (ULO) was used to increase CL number in rats and
determine whether this would compromise luteal tissue blood flow, oxygen
consumption and progestin secretion. All investigations were performed
in vivo, using a venous outflow technique on Day 16 of
gestation, when progesterone secretion is maximal. ULO, performed before
pregnancy, doubled CL number and total CL mass in the remaining ovary of six
treated compared to five control rats. Growth of CL was not affected. The rate
of ovarian blood flow (µL min−1 mg
CL−1) fell to 47% of control levels in ULO
animals and progesterone secretion (µg h−1
mg CL−1) to 68%. Secretion of the minor
progestin, 20α-hydroxypregn-4-en-one was not affected. Tissue oxygen
consumption was maintained despite the reduction in blood flow by an increase
in oxygen extraction from arterial blood. These results suggest that
overcrowding of CL in ULO-stimulated rat ovaries compromises luteal tissue
blood flow and subsequently progesterone secretion.
Luteinizing hormone receptors in the bovine corpus luteum during the oestrous cycle and pregnancy
Authors: Okuda K. , Uenoyama Y. , Naito C. , Sakabe Y. , Kawate N.
Published: November 2003

The concentration and affinity of luteinizing hormone (LH) receptors in bovine
luteal tissues during the oestrous cycle and pregnancy were investigated by
Scatchard analysis of the binding of 125I-labeled human
chorionic gonadotropin. Corpora lutea (CL) were classified into five stages of
the oestrous cycle and three stages of pregnancy. The concentration of LH
receptors sharply increased from the early I stage of the oestrous cycle (Days
2–3; 3.09 fmol mg-1 protein) to the early II stage
(Days 5–6; 9.44 fmol mg-1 protein) and then
remained constant until the late luteal stage (Days 15–17;
8.14–9.56 fmol mg-1 protein). The LH receptors
could not be analysed in the regressed luteal tissue due to the small amounts
of binding. There was no significant difference in the concentrations of LH
receptors (5.63–9.64 fmol mg-1 protein) among the
three stages of pregnancy. Moreover, the concentrations of the receptors in
the CL of pregnancy were comparable to those in the mid-cycle CL. The binding
affinity did not change significantly during the oestrous cycle and pregnancy.
Based on these results, it is assumed that the luteal function during the
entire period of pregnancy might be regulated, at least in part, by LH, which
is mediated via its specific receptors, and that the luteal function during
pregnancy seems not to be regulated by changes in the binding capacity and
affinity of LH receptors. To understand the physiological roles of LH in
regulating luteal function in pregnant cows, further studies are required.
Effects of hyperthermia on uterine blood flow and shunting through uterine arteriovenous anastomoses in the late-pregnant ewe
Authors: Miller Suzanne L. , Jenkin Graham , Walker David W.
Published: November 2003

The effect of maternal hyperthermia on uterine
blood flow (UBF) through the two main uterine arteries and on the proportion
of UBF shunted through uterine arteriovenous anastomoses (AVAs) was
investigated. Eight late-pregnant ewes were exposed to normothermic
(22–23˚C) or hyperthermic (approx-imately 39˚C) ambient
conditions for 8 h. UBF was measured in the left and right uterine arteries
using flow probes and microspheres were injected into the uterine artery
before, during and after the experimental period. The distribution of
microspheres between the uterus and lungs was determined to calculate changes
in capillary and AVA blood flows. Hyperthermia produced a significant
(P<0.05) increase in maternal core temperature
(+1.5˚C), increase in maternal blood pH (+0.21;
P<0.05) and decrease in maternal pCO
2 (–16.2 mmHg; P<0.05).
Blood flow to the uterine horn ipsilateral to the corpus luteum (CL) remained
unchanged during hyperthermia, whereas total UBF and blood flow to the
contralateral uterine horn were significantly decreased
(P<0.05), by 23.1% and 20.8%,
respectively, of pre-heat control values. The proportion of UBF shunted
through uterine AVAs during hyperthermia was not significantly different from
values observed in normothermic ewes (21.9 0.7%). Mild to moderate
hyperthermia in late-pregnant sheep induces respiratory alkalosis and
decreases total blood flow to the uterus, brought about by a decrease in blood
flow to the uterine horn contralateral, but not ipsilateral to the CL. Heat
treatment does not alter the proportion of UBF traversing uterine AVAs.
Cyclopiazonic acid, an inhibitor of calcium-dependent ATPases, induces exit from metaphase I arrest in growing pig oocytes
Authors: Petr J. , Rozinek J. , Vanourková Z. , Jílek F.
Published: November 2003

Calcium plays an important role in the
regulation of meiotic maturation in mammalian oocytes. In the present study,
mycotoxin cyclopiazonic acid (CPA), an inhibitor of calcium-dependent ATPases,
was used to mobilize intracellular calcium deposits in growing pig oocytes,
which had not attained full meiotic competence and in which maturation is thus
spontaneously blocked at the metaphase I stage. CPA treatment significantly
increased the ratio of growing oocytes that are able to overcome the
spontaneously occurring metaphase I block to complete their maturation at the
metaphase II stage. CPA treatment of a least 2 hours’ duration is
necessary to overcome the metaphase I block in growing oocytes. A similar
effect upon release from the spontaneous meiotic block at the metaphase I
stage was observed after treatment of growing pig oocytes with thapsigargin,
another inhibitor of endogenous calcium-dependent ATPases. Numerous calcium
deposits in vacuoles, the mitochondria and on the surface of yolk granules in
growing pig oocytes were observed. CPA treatment is able to mobilize calcium
from the mitochondria, but deposits in vacuoles and deposits on the surface of
yolk granules seem to remain intact after CPA treatment. A microinjection of
heparin, which is known to bind with the inositol trisphosphate receptors,
significantly decreased the ratio of CPA-treated growing oocytes overcoming
the block at the metaphase I stage. This indicates that CPA might mobilize
calcium in growing pig oocytes through inositol trisphosphate receptors. On
the other hand, a microinjection of procaine or a microinjection of ruthenium
red, both inhibitors of ryanodine receptors, did not prevent the overcoming of
the metaphase I block, induced by CPA treatment. The calcium channel blocker,
verapamil, significantly reduces the proportion of CPA-treated growing oocytes
that overcome the metaphase I block. This indicates that the influx of calcium
from extracellular sources is necessary to over-come the metaphase I block.
The calmodulin inhibitors ophiobolin A and W7 also reduce the proportion of
CPA-treated growing oocytes overcoming the metaphase I block.
Reduction in wool follicles prior to birth in Merino sheep
Authors: Edwards J. E. Hocking
Published: November 2003

This study was undertaken to identify whether
all secondary follicles that are initiated are present at birth in the Merino
fetus, and if not, when does net initiation of secondary follicles cease. Skin
was sampled from fetal lambs at 36, 26 and 16 days before the estimated date
of parturition and from lambs at birth and 2, 4, 6, 8 and 13 weeks after
birth. The ratio of secondary to primary follicles (S/P) reached a maximum
16 days before birth and was significantly lower at birth
(P<0.002) and at all postnatal ages
(P<0.05). There was no difference between S/P at
birth and S/P at later ages. Postnatal primary follicle density, secondary
follicle density and the percentage of fibre-producing follicles followed
similar patterns to those reported by others. This is the first conclusive
demonstration that secondary follicle initiation is completed several weeks
prior to birth and that there are less secondary follicles at birth than at
134 days of gestation.
Chronic cold exposure has no effect on brown adipose tissue in newborn lambs born to well-fed ewes
Authors: Gate J. J. , Clarke L. , Lomax M. A. , Symonds M. E.
Published: November 2003

It has been previously shown in twin-bearing ewes fed only 60% of their
metabolizable energy requirements for late pregnancy that chronic cold
exposure induced by winter shearing of the ewes results in larger lambs with
more brown adipose tissue. This effect appears to be primarily due to
prevention of a decline in fetal body and tissue weights between 145
days’ gestation and 2 h after birth (i.e. 147 days’ gestation) in
lambs born to underfed shorn ewes. The present study therefore examined the
impact, in ewes that were well fed (i.e. received 100% of their
metabolizable energy requirements) during the final month of gestation, of
chronic cold exposure induced by winter shearing on lamb birthweight and
perirenal adipose tissue composition as measured 2 h after birth. Perirenal
adipose tissue was analysed for its thermogenic activity (i.e. GDP binding to
mitochondria) and catecholamine content. These observations were combined with
similar measurements made in near-term (i.e. 145 days’ gestation)
fetuses sampled from well-fed unshorn ewes. There was no difference between
lambs born to shorn or unshorn ewes with respect to birth-weight or perirenal
adipose tissue weight and composition. Perirenal adipose tissue weight was
higher in lambs born to unshorn ewes than in fetuses. The thermogenic activity
of adipose tissue was 2-fold higher in lambs born to unshorn ewes compared
with 145-day-old fetuses. Epinephrine was detectable only at very low levels
in fetal perirenal adipose tissue, increasing 10-fold after birth, with no
difference between lambs born to shorn or unshorn ewes. In newborn lambs,
plasma growth hormone concentration was lower and insulin concentration higher
in shorn compared with unshorn groups. In conclusion, chronic cold exposure
induced by winter shearing had no effect on brown adipose tissue development
in lambs born to well-fed ewes.
An ultrasound-guided procedure to administer a label of DNA synthesis into fetal sheep
Authors: Greenwood Paul L. , Slepetis Ramona M. , Bell Alan W. , Hermanson John W.
Published: November 2003

A novel technique was developed to deliver a
bolus dose of a DNA label into the peritoneal cavity of fetal sheep at
85–130 days gestation. Use of markers to identify the site of injection
in fetuses from litters up to quadruplets, and immunohistochemistry to detect
the DNA label, 5-bromo-2¢-deoxyuridine (BrdU), confirmed the procedure
was successful in 85% of cases. Duration of the procedure was (mean SD)
44 16 min, and recovery from anaesthesia was rapid and uneventful in all
cases. Fetal weight was estimated with a high degree of accuracy (residual
standard deviation (RSD) = 297 g and r
2 = 0.93, P<0.001) and
the dose of label administered (110 33 mg BrdU/kg fetal weight) was
adequate in all cases. BrdU detected in fetal nuclei following injection into
amniotic fluid highlights the need for positive identification of the
injection site in timed, short-term studies, and suggests potential to further
develop the technique to investigate cellular events in fetal sheep younger
than 85 days of gestation. The results demonstrate that the procedure can be
used to determine in vivo whether or not nuclei have
entered the S-phase of the cell cycle.
Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro
Authors: Harayama Hiroshi , Miyake Masashi , Magargee Susan F. , Kunze Elaine , Kato Seishiro , Shidara Osamu , Iwamoto Eiji , Arikawa Satoko , Hammerstedt Roy H.
Published: November 2003

This study is a detailed investigation of
changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa
during an incubation designed to promote capacitation
in vitro. Ejaculated spermatozoa were collected from six
mature boars, washed, and incubated to promote capacitation. Sperm samples
were subjected to Western blotting-densitometric analyses, flow cytometry
after immunostaining and immunocytochemical observation by indirect
immunofluorescence. An antiserum to anti-agglutinin was raised in a rabbit by
subcutaneous injection of a purified antigen, as described previously
(Harayama et al. 1999). Western blotting-densitometric
analyses revealed an approximate halving of the amount of sperm-bound
anti-agglutinin during the first 45-min incubation, followed by a gradual
decrease thereafter. Comparison between immunostained sperm samples by flow
cytometry before and after incubation confirmed this decrease in sperm-bound
anti-agglutinin during the incubation. Microscopic characterization
established that this decrease occurred mainly on the acrosome.
Supplementation with seminal plasma (5% or 10%, v/v)
attenuated the decrease. These findings are consistent with the conclusion
that a large portion of the anti-agglutinin bound to sperm acrosomes is
released at an early stage of the capacitation process
in vitro.
Immunocytochemical localization of oestrogen receptors in perichondrium of antlers in red deer (Cervus elaphus)
Authors: Barrell G. K. , Davies R. , Bailey C. I.
Published: November 2003

Remodelling of cancellous bone of antlers to compact bone can be stimulated by
administra-tion of oestrogens and previous work has demonstrated the presence
of specific oestrogen binding in this tissue. In this study the presence of
oestrogen receptors (ER) in antler tissue from red deer males was exam-ined by
immunocytochemistry using a monoclonal mouse anti-human ER serum. Strong
positive staining was detected in the tip regions of immature antlers and was
confined primarily to the cells forming a fibrous layer of the perichondrium.
This finding indicates that the effects of oestrogens on remodelling of
cancel-lous bone of the antler may be mediated indirectly by the surrounding
connective tissue layers.
Expression of intercellular adhesion molecule-1 (ICAM-1) in choriodecidua with labour and delivery at term and preterm
Authors: Marvin Keith W. , Keelan Jeffrey A. , Sato Timothy A. , Coleman Matthew A. G. , McCowan Lesley M. E. , Mitchell Murray D.
Published: November 2003

To evaluate the association between
intercellular adhesion molecule-1 (ICAM-1) in the chorio-decidua and preterm
labour and delivery, ICAM-1 mRNA abundance was assessed by northern analysis,
and protein levels by ELISA, in samples of this tissue after term and preterm
delivery. The median ICAM-1 mRNA expression following preterm delivery (PTD)
was 4.8 and 3.8 times (P<0.05), respectively, those
following elective Caesarean section prior to labour at term (CST) and
following vaginal delivery after spontaneous labour at term (SLT). The
concentration of ICAM-1 protein in the PTD samples was 2.2 and 3.0 times
(P<0.05) those in CST and SLT samples, respectively.
The differences between the term groups were not significant. The results were
substantially the same when a preterm spontaneous labour (PTL) subgroup,
exclusive of deliveries complicated by pre-eclampsia or intrauterine growth
restriction, was compared with the term groups. Choriodecidual ICAM-1 mRNA
expression, but not ICAM-1 protein concentration, significantly correlated to
the degree of leukocyte infiltration of the PTD gestational membranes. Neither
correlated significantly to clinical indications of intrauterine or neonatal
infection. These findings indicate that ICAM-1 is expressed by the human
choriodecidua and that this expression is elevated with preterm labour and
delivery, particularly with increased leukocyte infiltration.
Timing and regulatory aspects of oocyte maturation in vitro in the tammar wallaby (Macropus eugenii)
Authors: Mate Karen E. , Buist Janine M.
Published: November 2003

Oocytes from a marsupial, the tammar wallaby
(Macropus eugenii), resemble those of eutherian mammals
in their ability to resume meiosis in vitro when cultured under suitable
conditions. Culture for 42–48 h in Eagle’s minimum essential
medium (EMEM) supplemented with 10% fetal calf serum, and 10 g mL
–1 porcine luteinizing hormone (pLH) was required
in order for oocytes, collected from the large antral follicles (> 2 mm
diameter) of tammar wallabies (primed with 6 mg of porcine follicle
stimulating hormone twice daily for four days), to proceed to metaphase II
(MII) of meiosis. Under these conditions, chromatin condensation was observed
within 4–8 h of culture in 61% of oocytes; metaphase I (MI)
chromosomes were observed from 18–30 h of culture (66%); and most
oocytes (76%) progressed to MII by 42 h in vitro.
The addition of cycloheximide, a protein synthesis inhibitor, at
concentrations of 1–100 g mL –1 , prevented
maturation of tammar wallaby oocytes in vitro. This
effect was reversible, as oocytes washed free of cycloheximide after 4 h of
incubation were able to progress to MII. The addition of cycloheximide to
wallaby oocytes at MI of meiosis prevented normal progression to MII
suggesting that proteins critical for nuclear maturation are synthesized
throughout the maturation process. Genistein, a protein kinase inhibitor
decreased maturation of wallaby oocytes in a dose dependent manner. However,
the concentration required to significantly inhibit maturation of wallaby
oocytes (60 g mL –1 ) was greater than that
required for eutherian species. Most wallaby oocytes were able to undergo
germinal vesicle breakdown (GVBD) in the presence of high concentrations of
genistein but produced abnormal chromatin configurations and were unable to
progress to MII. Future studies will examine whether cytoplasmic changes occur
in marsupial oocytes in vitro and their temporal
relationship to nuclear maturation.
Antigen-specific systemic and reproductive tract antibodies in foxes immunized with Salmonella typhimuriumexpressing bacterial and sperm proteins
Authors: de Jersey James , Bird Peter H. , Verma Naresh K. , Bradley Mark P.
Published: November 2003

Attenuated
Salmonella typhimurium strains are potential
‘safe’ delivery vectors of an oral immunocontraceptive vaccine for
the European red fox (Vulpes vulpes). In the present
study, model bacterial (Escherichia coli heat-labile
enterotoxin B subunit, LTB) and fox sperm (fSP10) antigens were expressed in
S. typhimurium SL3261 (DaroA)
under the control of the trc promoter. Adult female
foxes were given three oral immunizations with SL3261 containing either LTB
(SL3261/pLTB), fSP10 (SL3261/pFSP10) or a control plasmid (pKK233-2 or
pTrc99A). All foxes raised serum (IgG) and vaginal (IgG and IgA) antibodies
against S. typhimurium lipopolysaccharide (LPS). Each
fox that received SL3261/pLTB raised high titre LTB-specific serum and
vaginal IgG antibodies. However, only one of four foxes immunized with
SL3261/pFSP10 raised an anti-fSP10 immune response, in the form of low
titre serum and vaginal IgG antibodies. No vaginal IgA antibodies were raised
against either LTB or fSP10 in these experiments. The immune responses against
recombinant LTB and fSP10 resulted chiefly from the initial dose of antigen in
the inocula and were minimally influenced by continued
in vivo antigen expression. This study demonstrates for
the first time in the female red fox that oral
Salmonella can elicit specific systemic and reproductive
tract antibodies against heterologous, recombinant proteins.
Maternal behaviour and the survival of lambs in superfine wool sheep
Authors: Kuchel R. C. , Lindsay D. R.
Published: November 2003

Superfine merino lambs from superfine and medium-wool ewes were compared to
test the hypothesis that superfine ewes have poorer maternal capacity than
broader wool ewes. All lambs were sired by one ram using artificial
insemination in the superfine ewes and embryo transfer in the medium-wool
ewes. This ensured a similar genetic background in all lambs. Single born
lambs from the superfine ewes were 1.5 kg lighter than those from medium-wool
ewes (3.3 v. 4.8 kg, P<0.05).
Compared with the medium-wool ewes, in the first hour after birth the
superfine ewes spent less time grooming their lambs (8.7
v. 25.2 min, P<0.05) and more
time separated from them (21.3 v. 0.0 min,
P<0.05). The lambs of superfine ewes also stood and
attempted to suck for less time (17.7 and 2.4 min v.
32.4 and 10.5 min, P<0.05) than those of medium-wool
ewes. Twenty-nine per cent of lambs from superfine ewes died in the first 4
weeks after birth compared with only 4% of lambs from medium-wool ewes.
It was concluded that superfine ewes lost more lambs than medium-wool ewes and
that this was associated with their lambs being smaller at birth and with the
ewes being poorer mothers.
Effects of luteectomy on the maintenance of pregnancy, circulating progesterone concentrations and lambing performance in sheep
Authors: Al-Gubory K. H. , Solari A. , Mirman B.
Published: November 2003

In sheep, there have been few and conflicting
data regarding the necessity of the corpus luteum (CL) for the maintenance of
pregnancy. The aims of the present study were to examine the effect of
luteectomy on and after Day 50 of pregnancy on maternal plasma progesterone
concentrations and the progression of pregnancy, to determine the minimum
placental progesterone support required for the maintenance of pregnancy, and
to evaluate the effect of luteectomy on lambing performance. In Experiment 1,
four ewes luteectomized on Day 50 of pregnancy aborted 2–7 days after
surgery, whereas pregnancy progressed and parturition occurred between Days
143 and 149, with live lambs, in three of four ewes and in four ewes
luteectomized on Days 60 and 70 of pregnancy respectively. The mean ( SEM)
progesterone concentrations on the day before and one day after luteectomy
decreased from 4.87 0.85 to 0.42 0.06 ng mL –1
(P<0.01), from 4.57 0.51 to 0.80 0.12 ng mL
–1 (P<0.02) and from 6.05
0.52 to 1.67 0.11 ng mL –1
(P<0.01), respectively, for the ewes luteectomized on
Days 50, 60 and 70 of pregnancy. The fall in progesterone concentrations was
90%, 80% and 71%, respectively, for the ewes
luteectomized on Days 50, 60 and 70 of pregnancy. In Experiment 2, pregnancy
progressed in four ewes luteectomized on Day 70 and parturition occurred
between Days 146 and 149, with live lambs. The mean progesterone
concentrations declined (P<0.01) from 6.9 0.7 ng mL
–1 on the day before luteectomy to 2.1 0.3 ng mL
–1 the day after surgery. The concentrations of
progesterone in blood collected every 3 h during a 24-h period were stable on
Days 60 and 80 of pregnancy, but they were lower
(P<0.03) on Day 80 than on Day 60 of pregnancy, for
each time period examined. In Experiment 3, the gestation length and
birthweights of single, twin and triplet lambs were not different between the
control intact ewes (n = 111) and the ewes
luteectomized on Days 70–80 of pregnancy (n
= 71). Lamb mortality was not different between the two groups
(7.2% v. 8.4%, control v. luteectomized).
In conclusion, these results showed that (1) the sheep
CL is necessary to maintain pregnancy until at least Day 60,
(2) progesterone withdrawal induced by luteectomy on and
after Day 50 of pregnancy must be of a critical magnitude to provoke abortion,
(3) after Day 60 of pregnancy, the CL and the placenta
together secrete more progesterone than required for pregnancy maintenance,
(4) there is no apparent 24- hour rhythm in maternal
plasma progesterone concentrations before and after luteectomy, and
(5) luteectomy at mid pregnancy has no apparent effect
on gestation length, lamb birthweight or lamb mortality.
Effects of maternal captopril treatment on growth, blood glucose and plasma insulin in the fetal spontaneously hypertensive rat
Authors: Lewis R. M. , Vickers M. H. , Batchelor D. C. , Bassett N. S , Johnston B. M. , Skinner S. J. M.
Published: November 2003

In the spontaneously hypertensive rat (SHR) fetal growth and metabolism are
abnormal. It has been speculated that maternal hypertension may be the cause
of these abnormalities. Captopril treatment, which reduces maternal blood
pressure, during pregnancy and lactation, is reported to have a beneficial
effect postnatally, normalizing the blood pressure of offspring in the SHR. In
the present study, the effects of maternal captopril treatment on fetal growth
and plasma metabolites were investigated in the fetuses of two rat strains
(SHR and Wistar-Kyoto (WKY)), in order to determine whether normalizing
maternal blood pressure also normalized abnormalities in fetal growth and
metabolism. On fetal Day 20, SHR fetuses were lighter and placentae were
heavier than for the corresponding WKY. Captopril had no effect on fetal
weight in the SHR, but decreased it in the WKY. There was no effect of
captopril on placental weight. Fetal plasma insulin levels were higher in the
SHR than in the WKY and were decreased by captopril treatment in both strains.
Fetal blood glucose was elevated and fetal blood lactate was decreased in
captopril-treated litters from both strains. Captopril had no effect on fetal
plasma IGF-1 but fetal plasma IGF-2 levels were lower in the captopril-treated
SHR than in the captopril-treated WKY. These findings suggest that maternal
captopril treatment decreases insulin secretion in the fetal rat. High levels
of fetal plasma insulin suggest that the SHR fetus is insulin resistant. Fetal
insulin levels may contribute to the adverse consequences of gestational
captopril treatment observed in many species. The differences in the effect of
captopril on the two strains suggest that there are underlying endocrine
differences in the SHR.
Characterization of the expression of the alternative splicing of the ED-A, ED-B and V regions of fibronectin mRNA in bovine ovarian follicles and corpora lutea
Authors: De Candia L. M. , Rodgers R. J.
Published: November 2003

Fibronectin is an extracellular matrix
glycoprotein. Alternative splicing of fibronectin mRNA within three specific
regions, the extra domains (ED) A and B and the variable (V) or IIICS region,
result in the production of different isoforms of fibronectin. These isoforms
differentially regulate tissue developmental processes, such as those
occurring during follicular and luteal development. The specific isoforms of
fibronectin present in follicles and corpora lutea have not been identified.
To identify these, primers for reverse transcription polymerase chain reaction
(RT-PCR) were designed to the known bovine amino acid sequence of exons
flanking the ED-A, ED-B and V regions. PCR products from bovine fetal liver
cDNA were determined to be bovine fibronectin by the correct product size and
DNA sequence homology to other species, and to the known bovine amino acid
sequence. Bovine ovarian follicles (0.5–9 mm diameter) and corpora lutea
(cyclic, early to late mid-luteal phase) were shown to express ED-A+,
ED-A–, ED-B+, ED-B–, V+ and V– fibronectin
isoforms, similar to the liver, lung and kidney of fetuses, but generally not
of adult animals. Thus follicles and corpora lutea express isoforms of
fibronectin usually expressed in developing tissues.
Extra keyword: cDNA sequence.
Effects of progesterone on expression of messenger RNA encoding oxytocin-neurophysin, oxytocin receptor and prostaglandin G/H synthase-1 and -2 during the early oestrous cycle in the ovine corpus luteum
Authors: Al-Matubsi H. Y. , Frazer S. , Fairclough R. J. , Jenkin G.
Published: November 2003

This study was conducted to determine whether early progesterone treatment
plays a role in the regulation of messenger RNA (mRNA) expression for
oxytocin–neurophysin, oxytocin receptor, prostaglandin G/H synthase
(PGHS)-1 and PGHS-2 in the ovine corpus luteum. The expression of ovarian
oxytocin, oxytocin receptor, PGHS-1 and PGHS-2 mRNA was investigated in
control, progesterone- or RU486-treated ewes. Fifteen ewes were randomly
assigned to three groups to receive intramuscular injections of progesterone
(12.5 mg; n = 5), RU486, (2.5 mg
kg–1 bodyweight; n =
4) or corn oil (1 mL; n =6) twice daily from Day
1 to Day 3 post oestrus. On the morning of Day 4 post oestrus, the corpora
lutea were collected and analysed for oxytocin–neurophysin mRNA by
Northern blot using a labelled cDNA probe, and for the expressions of the
oxytocin receptor, PGHS-1 and PGHS-2 mRNA using the reverse transcription
polymerase chain reaction. Administration of progesterone or suppression of
progesterone activity with RU486 did not affect expression of
oxytocin–neurophysin mRNA (P>0.05).
Pretreatment of the ewes with progesterone resulted in the enhancement of
luteal oxytocin receptor mRNA expression and suppression of PGHS-1 and PGHS-2
mRNA (P<0.001). These results indicate that early
progesterone treatment does not control the expression of
oxytocin–neurophysin mRNA in the ovine ovary but may be involved in the
regulation of ovarian oxytocin receptor and PGHS expression. It is proposed,
on the basis of these results, that progesterone may play a role in premature
corpus luteum regression through an intra-ovarian mechanism involving the
induction of ovarian oxytocin receptor mRNA expression.
The generation and utilization of a cancer-oriented representation of the human transcriptome by using expressed sequence tags
Authors: Brentani Helena , Caballero Otávia L. , Camargo Anamaria A. , da Silva Aline M. , da Silva Wilson Araújo , Neto Emmanuel Dias , Grivet Marco , Gruber Arthur , Guimaraes Pedro Edson Moreira , Hide Winston , Iseli Christian , Jongeneel C. Victor , Kelso Janet , Nagai Maria Aparecida , Ojopi Elida Paula Benquique , Osorio Elisson C. , Reis Eduardo M. R. , Riggins Gregory J. , Simpson Andrew John George , de Souza Sandro , Stevenson Brian J. , Strausberg Robert L. , Tajara Eloiza H. , Verjovski-Almeida Sergio ,   The Human Cancer Genome Project Cancer Genome Anatomy Project Annotation Consortium*,   The Human Cancer Genome Project Sequencing Consortium†
Published: November 2003

Whereas genome sequencing defines the genetic potential of an organism,
transcript sequencing defines the utilization of this potential and links the
genome with most areas of biology. To exploit the information within the human
genome in the fight against cancer, we have deposited some two million
expressed sequence tags (ESTs) from human tumors and their corresponding
normal tissues in the public databases. The data currently define ≈23,500
genes, of which only ≈1,250 are still represented only by ESTs. Examination
of the EST coverage of known cancer-related (CR) genes reveals that <1% do
not have corresponding ESTs, indicating that the representation of genes
associated with commonly studied tumors is high. The careful recording of the
origin of all ESTs we have produced has enabled detailed definition of where
the genes they represent are expressed in the human body. More than 100,000
ESTs are available for seven tissues, indicating a surprising variability of
gene usage that has led to the discovery of a significant number of genes with
restricted expression, and that may thus be therapeutically useful. The ESTs
also reveal novel nonsynonymous germline variants (although the one-pass
nature of the data necessitates careful validation) and many alternatively
spliced transcripts. Although widely exploited by the scientific community,
vindicating our totally open source policy, the EST data generated still
provide extensive information that remains to be systematically explored, and
that may further facilitate progress toward both the understanding and
treatment of human cancers.
An emotion-induced retrograde amnesia in humans is amygdala- and β-adrenergic-dependent
Authors: Strange B. A. , Hurlemann R. , Dolan R. J.
Published: November 2003

The influence of emotion on human memory is associated with two contradictory effects in the form of either emotion-induced enhancements or decrements in memory. In a series of experiments involving single word presentation, we show that enhanced memory for emotional words is strongly coupled to decrements in memory for items preceding the emotional stimulus, an effect that is more pronounced in women. These memory effects would appear to depend on a common neurobiological substrate, in that enhancements and decrements are reversed by propranolol, a β-adrenergic antagonist, and abolished by selective bilateral amygdala damage. Thus, our findings suggest that amygdala-dependent β-adrenergic modulation of episodic encoding has costs as well as benefits.
Robust singular value decomposition analysis of microarray data
Authors: Liu Li , Hawkins Douglas M. , Ghosh Sujoy , Young S. Stanley
Published: November 2003

In microarray data there are a number of biological samples, each assessed for the level of gene expression for a typically large number of genes. There is a need to examine these data with statistical techniques to help discern possible patterns in the data. Our technique applies a combination of mathematical and statistical methods to progressively take the data set apart so that different aspects can be examined for both general patterns and very specific effects. Unfortunately, these data tables are often corrupted with extreme values (outliers), missing values, and non-normal distributions that preclude standard analysis. We develop a robust analysis method to address these problems. The benefits of this robust analysis will be both the understanding of large-scale shifts in gene effects and the isolation of particular sample-by-gene effects that might be either unusual interactions or the result of experimental flaws. Our method requires a single pass and does not resort to complex ”cleaning” or imputation of the data table before analysis. We illustrate the method with a commercial data set.