bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of mRNA Levels for the MHC Class I‐Like Molecules CD1 and FcRn in Preimplantation Mouse Embryos
Authors: WARNER CAROL M. , PASCHETTO MIRIAM G. 
Published: December 2003

PROBLEM: Major histocompatibility complex (MHC) antigens expressed on preimplantation embryos are important for the control of development, reproduction, and allo‐recognition of the embryo by the mother. Four types of MHC class I and MHC class I‐like antigens have recently been defined: class Ia, class Ib, class Ic, and class Id, based on their similar three‐dimensional protein structures. Class Ia and class Ib antigens are encoded in the MHC, whereas class Ic and class Id antigens are encoded by genes on other chromosomes. Both class Ia and class Ib MHC antigens are expressed on preimplantation mouse embryos. The function of the class Ia antigens on embryos is unknown, but the function of one class Ib antigen, Qa‐2, the product of the Ped gene, has been
found to control the rate of early cleavage division and subsequent
embryo survival. The expression of class Ic and class Id antigens on
preimplantation embryos has not yet been evaluated. In the present study,
we report the analysis of mRNA expression of two class Id genes,
CD1 and FcRn, in preimplantation mouse embryos.
METHOD OF STUDY: A reverse transcription‐polymerase chain reaction
(RT‐PCR) assay was performed to analyze mRNA levels for CD1 and
FcRn in 1‐cell, 2‐cell, 8‐cell, and blastocyst stage embryos from
C57BL/6 mice.
RESULTS: No expression of CD1 mRNA was found in any of the
preimplantation embryos tested. As a by‐product of this study, we found a
mistake in the published sequence of the mouse CD1 gene:
nucleotide 746 in the cDNA is a G not a C. This base change is in a site
recognized by the restriction enzyme PstI, thereby eliminating a
PstI cleavage site. Expression of mRNA for FcRn was found
in all preimplantation stages tested. Higher levels of mRNA for
FcRn were detectable in 2‐cell and 8‐cell embryos compared to
1‐cell and blastocyst stage embryos.
CONCLUSION: This study shows that mRNA for FcRn but not for
CD1 is found in preimplantation mouse embryos.
Fas‐Mediated Apoptosis of Mouse Embryo Stem Cells: Its Role During Embryonic Development
Authors: ZOU GANG‐MING , REZNIKOFF‐ETIÉVANT MARIE‐FRANÇOISE , MILLIEZ JACQUES , LÉON ANNE , VERGÉ VÉRONIQUE , HIRSCH FRANÇOIS 
Published: December 2003

PROBLEM: Fas antigen (APO‐1/CD95) can regulate the activity of various cells during adulthood. This study aimed at determining whether Fas may also be involved in the regulation of very early events such as the embryo preimplantation stage.  METHOD OF STUDY: We used mouse embryo stem (ES) cell line as a model for testing the effect of Fas crosslinking upon anti‐Fas monoclonal antibody (MoAb) treatment. In addition, this treatment was also applied to in‐vitro mouse‐embryo culture.
RESULTS: Flow‐cytometry analysis of cultured ES cells demonstrated an
increase in Fas expression, unchanged in the presence of mouse
interleukin‐2, while greatly upregulated in the presence of
lipopolysaccharide (LPS). As determined by various means, ES cells may
undergo a Fas‐mediated apoptosis, slightly but significantly intensified
by the addition of LPS to cell cultures. We also report that anti‐Fas
MoAb directly inhibited two‐cell stage mouse‐embryo (preimplantation)
development in in‐vitro culture conditions.  CONCLUSION: These data suggest a novel mechanism controlling the regulation of physiological cell turnover as well as blastocyst implantation in early embryo development.
Increased Apoptosis in Human Amnion is Associated with Labor at Term
Authors: HSU CHAUR‐DONG , MEADDOUGH ERIKA , BASHERRA HASAN , HARIRAH HASSAN , LU LI‐CHENG 
Published: December 2003

PROBLEM: To characterize whether increased apoptosis in human amnion was associated with labor at term.  METHOD OF STUDY: Human amnion were obtained from term patients with vaginal delivery (n=5) or who underwent elective Cesarean section
(C/S) without labor (n=5). Apoptosis was performed by the TUNEL
(Terminal dUTP Nuclear End Labeling) assay. All nucleated cells stained
with propidium iodide in the amnion epithelial cells were identified in
red fluorescence. TUNEL positive apoptotic nuclei were identified in
green fluorescence. Five random fields of each specimen were blindly
counted by investigators. The percentage of apoptotic nuclei of total
nuclei (apoptotic index) was calculated and compared between the two
groups (25 microscopic fields for each group, respectively).
RESULTS: Patients with term labor had a significantly higher mean
apoptotic index in amnion epithelial cells than that with elective C/S
without labor (27.3±4.1% versus 3.6±1.6%,
P<0.001).  CONCLUSIONS: Our data indicate that apoptosis in human amnion is significantly increased and associated with labor at term.
Fetal Endothelial Cells Express Vascular Cell Adhesion Molecule in the Setting of Chorioamnionitis
Authors: CRAVEN CATHERINE M. , WARD KEN , WARD KEN 
Published: December 2003

PROBLEM: In intrauterine infection, inflammatory mediators may be released into the fetal circulation prior to fetal infection. We hypothesize that, in chorioamnionitis, inflammation alters fetal blood vessels. To test this, fetal endothelial cells were examined for vascular cell adhesion molecule (VCAM).  METHOD OF STUDY: Umbilical cords (n=9) from placentas with
chorioamnionitis were immunostained for VCAM. Controls from preterm
preeclamptic pregnancies (n=7) without histologic inflammation
were selected, and matched for gestational age and method of delivery.
VCAM sections were reviewed by a pathologist blinded to clinical
diagnoses.
RESULTS: All endothelial cells from each of the nine cords from placentas
with chorioamnionitis had strong VCAM staining. Two of nine samples also
had acute cord vasculitis. No cord endothelial cells from preeclamptic
placentas demonstrated similar VCAM staining (p<0.01).  CONCLUSION: Histologic chorioamnionitis was associated with VCAM expression of the umbilical cord vessels. In chorioamnionitis, inflammatory mediators may have entered the fetal circulation to activate endothelial cells. Intrauterine inflammation was not restricted to the chorioamnion, but also involved the fetal circulation.
Production of Protein Hormones by Cultured Trophoblast Cells Isolated from Term and Early Placentae
Authors: MALEK ANTOINE , SAGER RUTH , WILLI AMARA , MÜLLER JÜRG , HÄNGGI WILLY , BERSINGER NIKLAUS , LEISER RUDOLF 
Published: December 2003

PROBLEM: To compare the capacity of de novo hormone synthesis
by cultured trophoblast cells isolated from early and term placenta as
cytotrophoblast, and to determine the ability of these cells to
proliferate in culture.
METHOD OF STUDY: Cytotrophoblast cells were isolated from term (TP,
38–42 weeks) and early placentae (EP, 8–13 weeks) by
enzymatic digestion and subsequent purification on a percoll gradient.
The net synthesis of the hormones human placental lactogen (hPL) and
human chorionic gonadotropin (hCG) was determined as the release during
culture+cell content after culture−cell content before culture.
Proliferation was determined using a dedicated colorimetric reagent
(CellTiter 96TM).
RESULTS: Using a percoll gradient we were able to isolate three cell
bands with densities of 1.051, 1.058, and 1.063 g/mL, which were
predominantly cytotrophoblast cells as shown by immunocytochemical
analysis. The cytotrophoblast cells with the highest density (1.063 g/mL)
were used because they were found to release the highest amount of
hormones and have shown the lowest rate of cell death after 6 days in
culture. Both hCG and hPL showed different patterns of release during the
first 2–3 days of culture between TP and EP. While the release by
EP cytotrophoblast cells continued during 6 days of culture (n=4),
the concentrations for TP cytotrophoblast (n=4) reached a plateau
between 4 and 6 days. Net de novo synthesis calculated for
3×104 TP trophoblast cells cultured for 6 days
(mean±SD, n=4) was 8.65±9.05 mU for hCG and
0.95±0.45 ng for hPL. For EP, it was 395.5±265.5 mU for hCG
and 148.8±84.2 ng for hPL. Net synthesis of hCG was>10‐fold (TP) and>70‐fold (EP) higher than the initial cell content. While at term, hPL synthesis was only a fraction of the initial cell content, production by EP cytotrophoblast was 106 times the initial cell content. The extent of cell death after 6 days in culture was significantly (
P<0.02) higher for term (30–40%) than for early
trophoblast (10–20%). Using a proliferation detection agent during
the first 3 days of culture with first trimester cytotrophoblast cells,
we did not find any changes in the proliferative activity.
CONCLUSIONS: There are differences in the functional activity between
trophoblast cells obtained from first and third trimester. The in
vitro findings are difficult to reconcile with the different patterns
of plasma concentrations of the two hormones observed in vivo during the course of pregnancy.
Does Leptin Exhibit Cytokine‐Like Properties in Tissues of Pregnancy?
Authors: SOH E.B.E. , MITCHELL M.D. , KEELAN J.A. 
Published: December 2003

PROBLEM: To determine whether leptin exhibits cytokine‐like properties in gestational tissues in light of its homologies with the class I family of cytokines.  METHOD OF STUDY: WISH and JEG3 cells, and amnion and choriodecidua explants, were treated inflammatory modulators (interleukin‐1β [IL‐1β], tumor necrosis factor‐α [TNF‐α] and bacterial lipopolysaccharide [LPS]) and leptin production was measured by immunoassay. Other agents known to regulate adipocyte leptin production were also tested for comparative purposes. In addition, WISH cells, JAR cells and placental explants were treated with leptin to assess its effects on production of IL‐8, IL‐6 and prostaglandin E2
(PGE2).
RESULTS: Leptin production by all cells and tissues studied was
unaffected by treatment with IL‐1β (2.5 ng/mL), TNF‐α (25
ng/mL) and LPS (2.5 μg/mL). Dexamethasone stimulated leptin production
over two‐fold by WISH and JEG3 cells, whereas insulin also stimulated a
two‐fold increase in leptin production in JEG3 cells. IL‐6 production by
JAR cells and placental explants was stimulated (two‐ to three‐fold) by
leptin (300 ng/mL). PGE2 production was unaffected.  CONCLUSIONS: Leptin derived from gestational tissues is unlikely to play a role in inflammatory reactions within the placenta, but may regulate placental cytokine production. The physiological significance of amnion‐derived leptin remains to be established.
Acute Intrauterine Infection Results in an Imbalance between Pro‐ and Anti‐Inflammatory Cytokines in the Pregnant Rabbit
Authors: K. LESLIE KIMBERLY , L. LEE SCOTT , M. WOODCOCK SUSAN , K. DAVIES JILL , L. ESKENS JOAN , S. GIBBS RONALD , HIRSCH EMMET , P. SHERMAN MICHAEL , S. MCDUFFIE JR ROBERT 
Published: December 2003

PROBLEM: Intrauterine infection results in an increase in cytokines. This study compared the time courses for the pro‐ and anti‐inflammatory cytokine responses in 33 pregnant rabbits at 70% gestation. Pro‐inflammatory markers were activated nuclear factor‐kappa B (NF‐κB) in placenta and tumor necrosis factor‐alpha (TNF‐α) in amniotic fluid. These were compared to the anti‐inflammatory cytokine, interleukin‐1 receptor antagonist (IL‐1ra), in placenta and uterus.  METHOD OF STUDY: Does were endoscopically inoculated with Escherichia
coli through their cervices and sacrificed at six intervals between 0
and 30 hr post‐inoculation.
RESULTS: Activated NF‐κB, determined by electromobility gel shift
assay, increased significantly 16 hr after bacterial inoculation (P≤0.05). This was directly mirrored by TNF‐α concentrations, determined by bioassay, in the amniotic fluid. However, IL‐1ra levels, determined by enzyme‐linked immunosorbent assay, did not increase in response to infection.  CONCLUSION: Intrauterine infection results in an imbalance between pro‐ and anti‐inflammatory cytokines that may potentiate infection‐induced preterm delivery.
Mammary Glands of Aly Mice: Developmental Changes and Lactation‐Related Expression of Specific Proteins, α‐Casein, GlyCAM‐1 and MAdCAM‐1
Authors: NISHIMURA TOSHIHIDE , KOIKE RYUJI , MIYASAKA MASAYUKI 
Published: December 2003

PROBLEM: (1) Aly mouse is an autosomal recessive mutant mouse
that lacks lymph nodes and Peyer's patches. Defects of mammary
glands might explain why these mice fail to nurture their pups. (2)
Expressions of glycosylation‐dependent cell adhesion molecule 1
(GlyCAM‐1) and mucosal addressin cell adhesion molecule‐1 (MAdCAM‐1) in
mammary glands of lactating mice have been suggested, but only scant
information is available.
METHODS: The thickness and weight of mammary glands were measured
directly. The expression of specific proteins, α‐casein, GlyCAM‐1,
and MAdCAM‐1 in mammary glands was detected by reverse
transcription‐polymerase chain reaction (RT‐PCR). MAdCAM‐1 expression was
also confirmed by immunohistochemistry (IHC).
RESULTS: Fat pads of mammary glands of homozygous (aly/aly)
mice were small and weighed less than those of heterozygous
(aly/+) mice. An increase in end‐bud development was evident in
aly/aly mice. During pregnancy, growth of the mammary gland
and alveolar formation were noted in both groups. The expression of
α‐casein, GlyCAM‐1 and MAdCAM‐1 was detected in both groups of mice
by RT‐PCR from late‐pregnancy. Immunohistochemically, MAdCAM‐1 was noted
in small vessels around the mammary globules.
CONCLUSIONS: Our results demonstrated marked changes in the structure of
the mammary glands of aly/aly mice. The expression of
lactation‐related proteins was detected during development, pregnancy,
and lactation in both aly/aly and aly/+ mice.
Pregnancy as a Model of Controlled Invasion Might Be Attributed to the Ratio of CD3/CD8 to CD56
Authors: ARCK P.C. , HAGEN E. , HILDEBRANDT M. , KLAPP B.F. , HERTWIG K. 
Published: December 2003

PROBLEM: Pregnancy can be considered as a model of successfully controlled tissue invasion. Cellular mediated immunity appears to regulate the controlled invasion of fetal trophoblast cells. In endometrium cancer, a dysregulation of invasive malignant cells can be observed. Since immuncompetent cells are known to be involved in recognition and rejection of ‘non‐self’ antigens, we investigated the presence and distribution pattern of CD3, CD8, CD56, and CD68 positive cells in decidua from normal and failing pregancies, compared with malignant and benign endometrium.  METHOD OF STUDY: Decidual tissue from first trimester normal pregancies (NP; n=15) and abortion (AB; n=12), endometrial samples
from premenopausal women (NE; n=8), and endometrioid
adenocarcinoma (EA; n=8) were examined by immunohistochemistry
using monoclonal antibody against large spectrum cytokeratin, and against
the receptors CD3, CD8, CD56 and CD68, respectively.
RESULTS: In NP, we observed 32.5% CD3, 44.7% CD56, and 22.9%
CD68+ cells. In AB, we found 36.9% CD3, 45.3% CD56, and 17.8%
CD68+ cells. The differences in ratio between normal pregnancy
and abortion were not statistically significant. In NE, we counted 39.5%
CD3, 30.2% CD56 and 30.2% CD68+ cells. In EA, we observed
47.9% CD3, 12.4% CD56 and 39.7% CD68+ cells. The decrease of CD56 positive cells in endometrioid adenocarcinoma was statistically significant. Interestingly, we found 4.1% of cells positive for CD8 in NP, 4.9% in AB, 22.7% in NE, and 48.2% in EA.  CONCLUSIONS: The increase of CD8 cells in NE, and particularely in EA, and decrease of CD56 cells, compared with NP or AB, suggests an interaction between CD8 cells and CD56 cells. Studying different pathological situations in the uterus, such as malignancies or ectopic pregnancies, might help us to understand the mechanisms involved in the maintenance of pregnancy.
Blocking Effect of Antisera to Recombinant Zona Pellucida Proteins (r‐ZPA) on In Vitro Fertilization
Authors: HASEGAWA AKIKO , KOYAMA KOJI , TSUBAMOTO HIROSHI , HAMADA YUKARI , KOYAMA KOJI 
Published: December 2003

PROBLEM: The zona pellucida is a good target antigen for contraceptive vaccines due to its strong immunogenicity and high tissue specificity. However, this contraceptive effect is inevitably associated with ovarian failure. Therefore, it is necessary to define an epitope of the zona antigen to which the antibody produced inhibits fertilization without any undesirable side effects.  METHOD OF STUDY: The DNA fragment coding for the NH2‐terminal
region of porcine zona pellucida proteins (ZPA) (1–198 amino acids)
and human ZPA (1–206 amino acids) was prepared to produce
recombinant porcine ZPA (r‐ZPA), r‐pZPA1–198 and
r‐hZPA1–206. Using Freund's complete adjuvant,
antisera against these proteins were raised in rabbits.
RESULTS: The resultant antisera to r‐pZPA1–198 and
r‐hZPA1–206 were cross‐reacted with each other on
enzyme‐linked immunosorbent assay and immunofluorescent staining. The
antiserum to r‐pZPA1–198 inhibited in vitro
fertilization in pigs, but not human sperm binding to the zona pellucida,
while the antiserum to r‐hZPA1–206 inhibited the human
sperm‐binding assay.
CONCLUSIONS: Antiserum to r‐ZPA inhibited fertilization in the animal
species possessing a homologous amino‐acid sequence as an immunogen. The
recombinant protein, r‐hZPA1–206 seems to be a feasible candidate for the development of contraceptive vaccines for humans.
Infection of Polarized Primary Epithelial Cells from Rat Uterus with Chlamydia trachomatis: Cell–Cell Interaction and Cytokine Secretion
Authors: KAUSHIC CHARU , GRANT KATIE , CRANE MARDI , WIRA CHARLES R. 
Published: December 2003

PROBLEM: The objective of this study was to examine the susceptibility of rat uterine epithelial cells (UEC) to infection with Chlamydia
trachomatis and to study the epithelial–stromal interactions
following infection.
METHOD OF STUDY: UEC were isolated from adult rats and grown in culture.
Polarized, confluent monolayers of UEC were infected with 106
IFU/well C. trachomatis (MoPn). In order to confirm infection,
MoPn was labeled with a fluorescent tracking dye, PKH‐26, and then used
in epithelial cell infections. Transepithelial resistances were measured
prior to and following infection to test the effect of Chlamydia
on the integrity of the epithelial monolayers. In other experiments,
polarized epithelial cultures were infected in the presence and absence
of stromal cells. Media was collected from the apical and basolateral
compartments of the cultures before and after infection and analyzed for
cytokines IL‐1α and TNF‐α.
RESULTS: Epithelial cell cultures infected with PKH‐26 labeled MoPn were
examined 4–5 days later. Bacterial inclusions were detected inside
epithelial cells indicating infection had occurred. Co‐localization of
PKH‐26 labeled bacteria with FITC‐labelled anti‐Chlamydia antibody
on the epithelial cells confirmed infection. No changes were found in
resistance across the monolayers of epithelial cells in the presence or
absence of infection. ELISA results indicate that UEC secrete IL‐1α
constitutively in vitro. Stromal cells secrete very little
IL‐1α. When stromal cells were co‐incubated with epithelial cells
there was a decrease in the amount of IL‐1α secreted by epithelial
cells 48 hr post‐infection. On the other hand, maximum TNF‐α was
found in stromal cells, both with and without infection. Epithelial
cells, in these studies made very little TNF‐α.
CONCLUSIONS: These results show that primary rat epithelial cells can be
infected with Chlamydia in vitro. Epithelial and stromal cells
from uteri of adult rats make IL‐1α and TNF‐αin vitro
both prior to and following infection with Chlamydia. This system can be used to analyze the role played by epithelial–stromal interactions in providing protection on this mucosal surface.
Autoimmunity in Endometriosis: Relevance to Infertility
Authors: MATHUR SUBBI P. 
Published: December 2003

PROBLEM: Endometriosis is a complicated multi‐factorial disease. Genetic and immunologic factors play a key role in its pathophysiology. We were the first to describe endometrial and ovarian autoimmunity in women with endometriosis in 1981.METHODS OF STUDY: ELISA and Western blot analysis. It is accepted that women with endometriosis have autoimmunity to endometrial, ovarian and nuclear antigens. We have now established that:

The endometrial autoantigens to which the endometriosis patients have autoimmune reactions are endometrial transferrin and alpha 2‐HS glycoprotein.
Levels of antibodies to both these proteins are specifically elevated in women with endometriosis, thus making them important candidates for developing an endometrial antibody assay for a non‐invasive diagnosis of endometriosis.
Levels of transferrin and alpha 2‐HS glycoprotein are significantly elevated in the peritoneal fluid of women with endometriosis.
Antibodies to transferrin and alpha 2‐HS glycoprotein also inhibit in vitro sperm motility.

CONCLUSION: These data strongly suggest the possibility of endometrial autoantibodies playing an important role in the infertility often associated with endometriosis.
Immunobiologic Implication of RANTES in Seminal Plasma of Fertile, Infertile and Immunoinfertile Men
Authors: NAZ RAJESH K. , LESLIE MONICA H. 
Published: December 2003

PROBLEM: To investigate the presence of Regulated on Activation, Normal T Expressed and Secreted (RANTES) in the seminal plasma of fertile, infertile, and immunoinfertile men in order to determine whether it has any role in fertility/infertility and in the modulation of immunogenicity of the sperm cell.  METHOD OF STUDY: The enzyme‐linked immunosorbent assay was performed by using a commercially available immunoassay kit to measure RANTES levels in seminal plasma of fertile (n=28), male‐factor infertile
(n=35), and immunoinfertile (n=21) patients. The levels
were correlated with various seminal parameters.
RESULTS: RANTES was detected in the seminal plasma of fertile,
male‐factor infertile, and immunoinfertile men. Its levels differed
significantly (P<0.05) between the fertile and immunoinfertile
groups, with the immunoinfertile group showing an overall decrease of
19.9% in mean values. When RANTES levels were correlated with sperm
concentration, total sperm number, and motility, only the total number of
the immunoinfertile group correlated (r=0.86) significantly
(P=0.03) with the RANTES levels, and none of the other seminal
parameters of any other group correlated
(r=−0.37–0.30) significantly (P>0.05).  CONCLUSIONS: The findings of this study indicate that RANTES may have a role in immunomodulation of antigenicity of sperm cells in the male genital tract before ejaculation, and perhaps also in the female genital tract after intercourse. The exact physiologic, pathopysiologic, and molecular mechanism(s) involved in these processes require further study.
Abnormal TGFβ Levels in the Amniotic Fluid of Down Syndrome Pregnancies
Authors: BROMAGE STEPHEN J. , LANG ALLISON K. , ATKINSON IAN , SEARLE ROGER F. 
Published: December 2003

PROBLEM: The status of cytokines in amniotic fluid (AF) from chromosomally abnormal pregnancy is largely undefined. TGFβ plays a key role in fetal growth and differentiation and is responsible for the immunoregulatory activity of AF in normal pregnancy, but its status in Down syndrome (DS) pregnancies is unknown. In addition we investigated the IL‐2 status of AF from DS pregnancies.  METHOD OF STUDY: Midtrimester AF from chromosomally normal (n=25)
and abnormal pregnancies with DS (n=15) were assayed for bioactive
and latent TGFβ levels using the mink lung epithelial cell growth
inhibition bioassay and for IL‐2 activity by the CTLL‐2 cell
proliferation bioassay and by ELISA.
RESULTS: Levels of bioactive TGFβ (mean 4.6±0.6 U/mL) were
significantly increased in DS AF compared with the normal samples (mean
2.8±0.3 U/mL, P<0.003) but latent TGFβ levels did
not differ between DS and normal groups. In addition DS AFs showed
reduced IL‐2 like bioactivity compared with normal samples
(P=0.0006) but IL‐2 immunoactivity was undetectable in DS and normal AFs by ELISA.  CONCLUSIONS: DS AFs display increased TGFβ activity and lacked IL‐2 immunoactivity. The reduced ability of DS AFs to stimulate CTLL‐2 cell proliferation is unrelated to the IL‐2 status of AF. Altered TGFβ levels may prove useful as an additional biochemical index for the detection of DS pregnancies.