bluuebunny/crossref_metadata_2025 huggingface dataset:
Risk of bias of animal studies on regenerative procedures for periodontal and peri‐implant bone defects – a systematic review
Authors: Faggion Clovis Mariano , Giannakopoulos Nikolaos Nikitas , Listl Stefan
Published: September 2011

Objectives

(1) To assess the risk of bias of studies in which animal models were used for investigating regenerative therapies for periodontal and peri‐implant defects. (2) To investigate changes in risk of bias by comparing samples drawn from two different publication periods.

Material & Methods

We searched the PubMed and LILACS electronic databases, independently and in duplicate, for randomized and controlled trials published from 1998 to 2000 and from 2008 to 2010. Hand searching included search of 10 dental journals, in the issues published between August 2008 and August 2010. Studies on non‐human primates and canines were included. We assessed independently and in triplicate the risk of bias with reference to a six‐item checklist based on the Cochrane Collaboration's tool for assessing the risk of bias and information about formal sample size calculation.

Results

One hundred and seven studies were included in the review. Checklist items were poorly reported in the studies selected, and therefore for most of the studies, the risk of bias was unclear.

Conclusion

As a result of the unclear risk of bias of animal studies in periodontal and peri‐implant treatments, it is difficult to determine the accuracy of treatment effect estimates. There is a need for standardization of reporting procedures on animal experiments.
Importance of life‐history and landscape characteristics for genetic structure and genetic diversity of brown trout (Salmo truttaL.)
Authors: Östergren Johan , Nilsson Jan
Published: September 2011

Abstract – Investigating the influence of evolutionary forces on the genetic structure and genetic diversity remains a major challenge. Yet, it is of considerable interest for conservation and management of a species. This study investigates the influence of life‐history and landscape features, such as altitude, connectivity and habitat size, on genetic diversity and genetic structure of brown trout (Salmo truttaL.) with stream‐resident, lake‐dwelling and sea‐migrating life‐history in two river systems in northern Sweden. Using regression tree analysis including ecological and landscape characteristics, we show that life history is the most important variable explaining genetic diversity and population differentiation. Sea‐migrating populations show high diversity and low differentiation, and lake‐ and stream‐resident populations show low diversity and high population differentiation, among all samples. No overall genetic correlation with geographical distance was noted; however, among sea‐migrating populations within the River Vindelälven drainage, this pattern was observed. This study illustrates that life‐history and landscape features help to explain genetic structure and genetic variation. The information is important for conservation and management actions, such as fisheries regulations, habitat restorations, stocking of hatchery fish, defining management units and introducing genetic monitoring programmes.
Anti‐human gonadotropin antibodies generated during in vitro fertilization (IVF)‐related cycles: Effect on fertility of rhesus macaques
Authors: Iliff Susan A. , Molskness Theodore A. , Stouffer Richard L.
Published: September 2011

Administration of human gonadotropins such as hFSH, hLH, and hCG to rhesus macaques can result in formation of anti‐human gonadotropin antibodies. To determine whether the presence of these antibodies interferes with subsequent fertility, sixteen female rhesus macaques (Macaca mulatta) with known antibody levels were bred with male rhesus macaques. The presence of antibodies did not interfere with conception or maintenance of pregnancy. Furthermore, antibody titers did not increase during gestation or following the resolution of pregnancy.
Normal serum biochemical and hematological values of the Sulawesi macaques
Authors: Goodrich J.A. , Ward G.S. , Swindle M.M.
Published: September 2011

Sulawesi macaque (SM) species are currently of interest to primatologists for genetic and behavioral studies world wide. However, there are no published reference hematological and serum biochemical profile values for several of the seven SM species. Twenty‐five clinically healthy, outdoor housed, ketamine sedated Macaca tonkeana and Macaca maurus were serially sampled (two to four times) over a 16‐month period (N = 68). Normal ranges were defined to include two standard deviations and 2.5–97.5 percentile. Significant differences in single sample date comparisons (P < 0.05) of species (serum Ca, MCHC), age (BUN, BUN/creatinine, alkaline phosphatase, phosphorus), gender (total protein, RBC, hemoglobin, HCT) and pregnancy (phosphorus and neutrophil count) groups were found. This normative data facilitates clinical evaluation of SM species and allows comparison to other macaque species.
Induction of immunodeficiency virus‐specific immune responses in rhesus monkeys following gene gun‐mediated DNA vaccination
Authors: Fuller Deborah Heydenburg , Murphey‐Corb Michael , Clements Janice , Barnett Susan , Haynes Joel R.
Published: September 2011

Abstract: The Accell® gene delivery system (gene gun) was used to deliver gold particles coated with HIV‐1LAI and SIVmac239 expression constructs into the epidermis of rhesus macaques, resulting in the elicitation of env‐ and gag‐specific humoral responses. One microgram of vector DNA per dose was sufficient to induce immune responses in monkeys using SIVmac239 gp160 and gp120 vectors driven by the CMV‐intron A promoter. Several parameters, including the identity of the vector, the length of the rest period between immunizations, the number of immunizations, and the amount of DNA per immunization, are all important in designing an optimal DNA immunization regimen. In addition, gene gun‐based DNA immunization using low efficiency expression vectors is an effective means of priming for the induction of vigorous antibody responses in macaques following boosting with recombinant subunits.
Safety and immunogenicity of intramuscular and intravaginal delivery of HIV‐1 DNA constructs to infant chimpanzees
Authors: Bagarazzi Mark L. , Boyer Jean D. , Javadian M. AM , Chattergoon Michael , Dang Kesen , Kim Grace , Shah Jyoti , Wang Bin , Weiner David B.
Published: September 2011

Abstract: Any global strategy for controlling the human immunodeficiency virus (HIV) epidemic is likely to rely heavily on immunization of infants and children. Given the well‐documented differences in children's responses to traditional vaccines, we initiated this study to extend our findings on DNA vaccination of adult chimpanzees to immunologically immature infant chimpanzees. Our findings were consistent with our previous work in adults as we observed that the DNA vaccines used here were both well tolerated and immunogenic within weeks of the initial vaccination.
Mechanisms of protection induced by attenuated simian immunodeficiency virus. IV. Protection against challenge with virus grown in autologous simian cells
Authors: Almond N. , Corcoran T. , Hull R. , Walker B. , Rose J. , Sangster R. , Silvera K. , Silvera R , Cranage M. , Rud E. , Stott E.J.
Published: September 2011

Abstract: Attenuated simian immunodeficiency virus (SIV) induces potent protection against infection with wild‐type virus, but the mechanism of this immunity remains obscure. Allogeneic antibodies, which arise within animals as a result of SIV infection, might protect against challenge with exogenous SIV grown in allogeneic cells. To test this hypothesis, eight macaques were infected with attenuated SIV and subsequently challenged with wild‐type SIV grown in autologous cells or heterologous cells.

The results clearly demonstrated that animals infected with attenuated SIV are protected against wild‐type SIV grown in autologous or heterologous cells. Thus, the hypothesis that live attenuated SIV protects by the induction of allogeneic antibodies is not tenable.
Chimeric SHIV that causes CD4+ T cell loss and AIDS in rhesus macaques
Authors: Joag Sanjay V. , Li Zhuang , Wang Chunyang , Jia Fenglan , Foresman Larry , Adany Istvan , Pinson David M. , Stephens Edward B. , Narayan Opendra
Published: September 2011

Abstract: By animal to animal passage in rhesus and pig‐tailed macaques, we developed a rhesus model of HIV‐1 disease in humans. Rhesus macaques infected with a cell‐free stock of SHIVKU‐2 developed CD4+ T cell loss, primary lentiviral encephalitis and pneumonia, and AIDS. Six of nine rhesus macaques died within eight months post‐inoculation, while the remaining three are at five, five, and eight months post‐inoculation, respectively. Animals infected by either mucosal or parenteral routes of infection had a similar course of infection.
In vitro infection of primate PBMC with simian/human immunodeficiency virus, SHIVSF33A: Correlation to in vivo outcome
Authors: Harouse Janet M. , Tan Rei Chin How , Gettie Agegnehu , Dailey Peter , Marx Preston A. , Luciw Paul A. , Cheng‐Mayer Cecilia
Published: September 2011

Abstract: The macaque/SIV animal system is an important model for studying AIDS pathogenesis and for evaluating the efficacy of vaccines and anti‐viral therapeutics. However, differences between HIV‐1 and SIV envelope proteins exist that render the SIV/macaque model of limited value when examining envelope determinants of retroviral pathogenesis. To overcome this problem, we utilized a chimeric virus, SHIVSF33, containing the env gene from HIV‐1SF33 in the context of the molecular clone SIVmac239, in the macaque animal model. In this study SHIVSF33A, a pathogenic virus that evolved in vivo from a rhesus macaque infected intravenously with the molecular clone SHIVSF33 was used in both in vitro and in vivo studies. By using a cell culture system, we examined the biological properties of our parental and animal‐adapted chimeric viruses and compared in vitro susceptibility to in vivo studies.
Opposing effects of ultraviolet B irradiation on interleukin‐1 receptor antagonist and interleukin‐1α messenger RNA expression in a human epidermoid carcinoma cell line
Authors: Lew W. , Lee S. H. , Park Y. K.
Published: September 2011

Interleukin‐1 receptor antagonist (IL‐1RA) is a cytokine that acts to antagonize IL‐1 activity without agonist function. The expression of IL‐1RA has been reported in many cell types, including the keratinocyte that covers the outer most part of the skin. However the modulation of IL‐1RA by ultraviolet B (UVB), which is the most biologically active UV, has not been reported yet. We therefore selected a keratinocyte cell line with a cytokine‐producing profile similar to that of keratinocytes and tested the effect of UVB on its ability to produce IL‐1RA mRNA. IL‐1RA mRNA was constitutively expressed in the cell line and began to be suppressed by 3 h after the UVB irradiation with 100 mJ/cm2. The level of IL‐1RA expression became lowest by 16 h after the irradiation with 100 mJ/cm2. Simultaneously, IL‐1α mRNA started to increase by 1 h and peaked by 3–16 h after the irradiation with 10–100 mJ/cm2. The differential expression of IL‐1α and IL‐1RA mRNA following exposure to a high dose (100 mJ/cm2) of UVB may markedly potentiate the role of IL‐1 in UV‐induced inflammation.
Neoplastic transformation of neonatal human fibroblasts exposed in vitro to radiation from a quartz‐halogen lamp
Authors: West R. W. , Rowland K. L. , Miller S. A. , Beer J. Z.
Published: September 2011

The use of unfiltered quartz‐halogen lamps exposes human skin to radiation that spans much of the ultraviolet (UV) spectrum. Reports indicate that exposure to quartz‐halogen lamps is erythemogenic, mutagenic, and carcinogenic. To compare the carcinogenic potential of quartz‐halogen lamps with that of other UV sources, we determined the dose dependence for cytotoxicity and neoplastic transformation in neonatal human fibroblasts exposed in vitro to: a 15 W germicidal lamp (primarily 254 nm radiation), a 15 W Cool White fluorescent lamp, and an unfiltered 20 W quartz‐halogen lamp. Fluence‐survival relationships were multiphasic with linear dose response below about 40% survival, and all three sources produced fluence‐dependent transformation as indicated by induction of anchorage‐independent growth. Maximum transformation frequencies were observed at fluences of 5–8 J/m2 for the germicidal lamp, 6.3 kJ/m2 for the fluorescent lamp, and 300 J/m2 for the quartz‐halogen lamp. These data confirm the carcinogenic potential of the quartz‐halogen lamp.
Long‐wave ultraviolet radiation causes increase of membrane‐bound fraction of protein kinase C in rat myeloid leukemia cells
Authors: Leszczynski D. , Leszczynski K. , Servomaa K.
Published: September 2011

We examined the effect of long‐wave ultraviolet radiation (UVA) on protein kinase C (PKC) and on the proliferation of rat myeloid leukemia cell line (ChL). Exposure of cells to a single dose of UVA (8 J/cm2 at 372± 10 nm) caused a rapid increase in the quantity of the membrane‐bound PKC, as assessed by 3H‐phorbol ester (3H‐PMA) binding assay (performed at 4°C). Within 2 h of UVA irradation, three peaks of increased 3H‐PMA binding to the ChL cells (by 70–100%) were observed at ca. 20, 60 and 95 min post‐irradiation. The exposure of ChL to UVA caused also a rapid, but transient, decline in the cell proliferation rate (by 18% within 24 h). However, the statistically significant decrease in cell numbers was observed only 3 days later (down by 22%). The inhibition of ChL proliferation was not due to alteration of cell viability as determined by trypan blue exclussion assay, and neither was it caused by cell cycle arrest or apoptosis, as determined by flow cytometry analysis of propidium iodide‐labelled cells and cell morphology in May‐Grünvald‐Giemsa‐stained cell smears. Phorbol‐ester‐induced activation of PKC (performed at 37°C) caused inhibition of ChL proliferation similar to that caused by UVA. This suggests that a UVA‐induced increase of the membrane‐bound fraction of PKC may be responsible for the UVA‐induced inhibition of ChL proliferation.
Photohemolytic potency of oral antidiabetic drugs in vitro: effects of antioxidants and a nitrogen atmosphere
Authors: Selvaag E.
Published: September 2011

The sulphonamide‐derived oral antidiabetic drugs carbutamide, chlorpropamide, glibenclamide, glibornuride, gliclazide, glipizide, gliquidone, glisoxepide, glymidine, tolazamide and tolbutamide were investigated for photohemolytic properties in vitro. Irradiation with a SOL 3 apparatus (solar simulating irradiation) revealed hemolysis in the presence of chlorpropamide, glipizide, gliquidone, glymidine and tolbutamide (all in the concentration 10−3 mol/l). Except for glymidine, which exerted photohemolysis in the concentration 10−4 mol/l, no hemolytic effects were seen in the concentration of 10−4 mol/l or 10−5 mol/l. Irradiation with TL 12 light bulbs (UVB), a UVASUN 5000 apparatus (UVA) or an experimental lamp (visible light) did not induce phototoxic hemolysis with either of the test substances. Addition of the antioxidants ascorbic acid, alpha‐tocopherol or superoxide dismutase significantly inhibited the phototoxic hemolysis. Investigations carried out in a nitrogen‐rich atmosphere reduced the hemolysis as well. These findings indicate an involvement of reactive oxygen species in the mechanism of action of the hemolytic process in the presence of oral antidiabetic drugs.
Effects of low concentrations of cis‐ and trans‐urocanic acid on cytokine elaboration by keratinocytes
Authors: Redondo P. , García‐Foncillas J. , Cuevillas F. , España A. , Quintanilla E.
Published: September 2011

The urocanic acid cis isomer (cis‐UCA) is a possible cutaneous photoreceptor for the immunomodulatory phenomena that follow ultraviolet B irradiation. Several experiments in animals show an inhibitory action of cis‐UCA on cellular immunity. However, the action of cis‐UCA on the synthesis of cytokines in keratinocytes remains unknown. Long‐term cultures of normal human keratinocytes were prepared in a serum‐free medium, and stimulated with 1 μg/ml of phorbol 12‐myristate 13‐acetate (TPA) and UCA or UVB‐UCA (10–100 μg/ml). Synthesis of the following cytokines was measured using ELISA and Northern blot techniques: TNF‐α, IL‐1α, IL‐1β, IL‐6, IL‐8 and TGF‐β1. TPA increased TNF‐α protein levels in culture supernatants. No changes in IL‐1α and IL‐1β protein levels were detected in basal culture supernatant after TPA stimulus. TPA augmented RNA expression for TNF‐α, IL‐1α, IL‐1β and TGF‐β1. UCA isomers did not induce cytokine changes in protein synthesis. Expression of IL‐1α and IL‐1β genes was increased after exposure to 100 μg/ml UVB‐UCA (70 μg/ml cis‐UCA). A slight increase in TNF‐α RNA expression was detected when the dose of UVB‐UCA reached 100 μg/ml. No effects on cytokine synthesis were found after UCA stimulus. These results suggest that low doses of cis‐UCA do not effect cytokine synthesis by keratinocytes.
Survey of the variation in ultraviolet outputs from ultraviolet A sunbeds in Bradford
Authors: Wright A. L. , Hart G. C. , Kernohan E. , Twentyman G.
Published: September 2011

Concerns have been expressed for some time regarding the growth of the cosmetic suntanning industry and the potential harmful effects resulting from these exposures. Recently published work has appeared to confirm a link between sunbed use and skin cancer. A previous survey in Oxford some years ago demonstrated significant output variations, and we have attempted to extend and update that work. Ultraviolet A, UVB and blue‐light output measurements were made on 50 sunbeds using a radiometer fitted with broad‐band filters and detectors. A number of irradiance measurements were made on each sunbed within each waveband so that the uniformity of the output could also be assessed. UVA outputs varied by a factor of 3, with a mean of 13.5 mW/cm2; UVB outputs varied by a factor of 60, with a mean of 19.2 μW/cm2; and blue‐light outputs varied by a factor of 2.5, with a mean of 2.5 mW/cm2. Outputs fall on average to 80% of the central value at either end of the sunbed. Facial units in sunbeds ranged in output between 18 and 45 mW/cm2. Output uniformity shows wide variation, with 16% of the sunbeds having an axial coefficient of variation > 10%. UVB output is highly tube‐specific. Eyewear used in sunbeds should also protect against blue light.
Psoralen‐fatty acid adducts activate melanocyte protein kinase C: a proposed mechanism for melanogenesis induced by 8‐methoxypsoralen and ultraviolet A light
Authors: Anthony F. A. , Laboda H. M. , Costlow M. E.
Published: September 2011

Diacylglycerol, a protein kinase C activator, induces and enhances melanogenesis in vitro and in vivo, providing evidence that melanogenesis may be a protein kinase C‐mediated process. Melanogenesis is also induced by ultraviolet A radiation and potentiated by a combination of 8‐methoxypsoralen and ultraviolet A radiation. We incubated cultured normal human melanocytes with 8‐methoxypsoralen, irradiated the cells with ultraviolet A radiation, and detected formation of 8‐methoxypsoralen‐phospholipid photoadducts. The 8‐methoxypsoralen‐phospholipid photoadducts isolated from melanocytes were substrates for phospholipase A2 to generate 8‐methoxypsoralen‐fatty acid adducts. We found that 8‐methoxypsoralen‐fatty acid photoadducts prepared in vitro could be substituted for diacylglycerol to activate protein kinase C in a cell‐free system. We propose that 8‐methoxypsoralen‐fatty acid adducts activate protein kinase C to potentiate ultraviolet A radiation‐induced melanogenesis. This proposal links melanogenesis mediated by protein kinase C with that induced by a combination of 8‐methoxypsoralen and ultraviolet A radiation.
What is actinic prurigo in Britain?
Authors: Grabczynska S. A. , Hawk J. L. M.
Published: September 2011

Actinic prurigo (AP) belongs to the group of idiopathic photodermatoses sharing a predilection for occurring more commonly in females, and there is much controversy as to whether it is only a more severe form of polymorphous light eruption (PMLE) or whether it is a distinct entity in its own right. The condition is characterised by intensely itchy papules, plaques and nodules, along with excoriations and scars usually starting before puberty, and predominantly involves the sun‐exposed areas although it may also affect covered sites. Seasonal exacerbations at the beginning of spring with improvement in the fall are typical, although the lesions frequently do not clear completely in the winter. The disorder may run a chronic course and persist into adulthood, but often spontaneous resolution occurs in late adolescence. Diagnosis is predominantly based on the clinical features, cutaneous irradiation tests and histology often being normal or non‐specific. HLA typing has also been performed in both PMLE and AP patients, showing a strong association between HLA‐DR4, in particular with the DRB1*0407 subtype, and AP; no HLA association has been found in PMLE. This HLA association is likely to be of pathogenic significance and strongly suggests a critical role for MHC‐restricted antigen presentation in the development of photosensitivity AP.
Effect of UVB 311 nm irradiation on normal human skin
Authors: Viac J. , Goujon C. , Misery L. , Staniek V. , Faure M. , Schmitt D. , Claudy A.
Published: September 2011

Ultraviolet radiation B (UVB) on the skin induces erythema, inflammation and modifications of the immune system. These changes have been reported after excessive short‐term or long‐term exposure to broad spectrum UVB. In this study, we examined the effects of local repetitive UVB irradiation of 311 nm wavelength on the skin of seven young volunteers. Skin biopsies were taken before and after UVB irradiation, and we immunohistochemically analyzed the expression of CD1a and HLA‐DR antigens of Langerhans cells (LC), the possible infiltration of dermis/epidermis by CD11b macrophages, the modifications or the induction of intercellular adhesion molecule‐1 (ICAM‐1), E‐selectin and vascular cell adhesion molecule‐1 (VCAM‐1) involved in the binding of leukocytes to the endothelial surface and the development of perivascular infiltrates of LFA‐1+ mononuclear cells. We also determined the expression of substance P receptors (SPR) using biotinylated substance P (SPB). Exposure of UVB 311 nm induced a drastic reduction of CD1a+ cells and a moderate increase of HLA‐DR+ dendritic cells in the epidermis without infiltration by CD11b macrophages. An increase of the binding of SPB to upper layer epidermal cells was noted in five of seven biopsies. In the dermis, vessel‐associated ICAM‐1 expression increased and an induction of E‐selectin occurred on nearly 20 to 40% of endothelial cells, but VCAM‐1 expression remained undetectable. The percentage of LFA‐1+ cells did not change significantly after irradiation. These observations may be compatible with a selective role of UVB 311 nm on the skin immune response.
Effects of frequency on gross efficiency and performance in roller ski skating
Authors: Leirdal S. , Sandbakk Ø. , Ettema G.
Published: September 2011

The purpose of the present study was to examine the effect of frequency on efficiency and performance during G3 roller ski skating. Eight well‐trained male cross‐country skiers performed three submaximal 5‐min speeds (10, 13, and 16 km/h) and a time‐to‐exhaustion (TTE) performance (at 20 km/h) using the G3 skating technique using freely chosen, high, and low frequency at all four speeds. All tests were done using roller skis on a large treadmill at 5% incline. Gross efficiency (GE) was calculated as power divided by metabolic rate. Power was calculated as the sum of power against frictional forces and power against gravity. Metabolic rate was calculated from oxygen consumption and blood lactate concentration. Freely chosen frequency increased from 60 to 70 strokes/min as speed increased from 10 to 20 km/h. GE increased with power. At high power (20 km/h performance test), both efficiency and performance were significantly reduced by high frequency. In regard to choice of frequency during G3 roller ski skating, cross‐country skiers seems to be self‐optimized both in relation to energy saving (efficiency) and performance (TTE).
Teratogenic IgG From Sera of Women With Spontaneous Abortions Seem to Induce Anomalies and Yolk Sac Damage in Rat Embryos. A Possible Method to Detect Abortions of Immunologic Origin
Authors: Abir Ronit , Ornoy Asher
Published: September 2011

PROBLEM: Spontaneous abortions due to immunological rejection of the embryo may be avoided by immunotherapy with paternal allogeneic leukocytes but there is no appropriate method to detect and differentiate this group of aborters from other groups.

METHODS: In previous studies we have demonstrated that in about two‐thirds of sera from women with spontaneous abortions the IgG antibodies are responsible (alone or in combination with other factors) for the embryotoxic effects of these sera on cultured rat embryos. We presently cultured 10.5‐day‐old rat embryos on highly teratogenic serum (“high risk” serum that induced anomalies in more than 50% of the embryos) from women with spontaneous abortions, where the IgG fraction was exchanged with IgG from control sera and vice‐versa. We studied by Transmission Electron Microscopy (TEM) the extent of yolk sac damage in comparison to the rate of embryonic anomalies.

RESULTS: In cases where IgG antibodies were teratogenic, embryos cultured in control sera with IgG from “high risk” sera exhibited ultrastructural yolk sac damage as well as embryonic anomalies, and the yolk sacs cultured in “high risk” sera with control IgG were normal. In cases in which the IgG exchange did not change the rate of anomalies, as IgG was not teratogenic, yolk sacs from embryos cultured in “high risk” sera remained damaged, while yolk sacs from embryos cultured in control sera after IgG exchange stayed normal. Although no significant difference in total IgG levels was found between the groups, a higher IgG1 level in sera from women with teratogenic IgG was observed in comparison to control women's sera. The obstetrical history of the women with two or more abortions who took part in our study showed that there were more cases of unknown etiology of the abortion in the women from the “high risk” group.

CONCLUSIONS: The serum and the IgG fraction from women with habitual abortions can be tested in whole embryo culture to evaluate the embryonic and yolk sac damage. On this basis it may be possible to detect the women in whom the habitual abortions result from immunological rejection.
Placental Expression of HLA Class I Genes
Authors: Le Bouteiller Philippe , Rodriguez Anne‐Marie , Mallet Valérie , Girr Maryse , Guillaudeux Thierry , Lenfant Françoise
Published: September 2011

This article presents an overview of the more recent data dealing with the constitutive, transcriptional, and translational expression of classical class la and nonclassical HLA‐E and ‐G class Ib products in the different trophoblast cell subpopulations that constitute the maternofetal interface during human pregnancy. Of particular interest is the expression of alternatively spliced HLA‐G transcriptional isoforms that may be translated in membrane‐bound or soluble protein products. Molecular regulatory mechanisms that may control the differential expression of class la and class lb molecules, according to the cell types, state of differentiation, and stages of gestation are also examined. They may operate at the levels of transcription, translation and/or transport of proteins to the cell surface. Functional significance of the absence of detectable cell surface expression of class Ia molecules in all trophoblast cell sub‐populations, and of the presence of membrane‐bound HLA‐G products in extravillous cytotrophoblast cells is finally questioned.
Effect of Pregnancy on Thymic T Cell Development
Authors: Ruhsinghani Asha G. , Bhatia Sudershan K. , Tygrett Lorraine T. , Waldschmidt Thomas J.
Published: September 2011

PROBLEM: The thymus gland decreases in size during pregnancy. The significance of this alteration is not known.

METHOD: In this report, we examined thymic function by evaluating the development of T lymphocytes in the thymus of pregnant Balb/c mice at 15 and 20 days gestation using multi‐color flow cytometry. Comparative analysis was made with non‐pregnant mice, postpartum lactating mice, and postpartum non‐lactating mice.

RESULTS: Progressive reduction of thymic size and cellularity during pregnancy was observed. All of the CD4 and CD8 defined subsets were reduced, with a disproportionate loss of CD4+, CD8+ double positive cells. Examination of the CD4‐, CD8‐ double negative compartment revealed a predominance of TCR α,β+ double negative cells, and a striking loss of precursor cells. The CD3‐, CD4‐, CD8‐ triple negative thymic subset was composed almost entirely of the earliest population (CD44+, CD25‐), with the remaining maturational stages (CD44+, CD25+; CD44‐, CD25+; and CD44‐, CD25‐) depleted. At 2 weeks postpartum, the subset ratios normalized, and the total cell count showed recovery.

CONCLUSION: T cell development is blocked at the precursor level during the mouse pregnancy. These effects are transient, and gradual recovery is observed in the postpartum period.
Antibodies to Endometrial Transferrin and Alpha 2‐Heremans Schmidt (HS) Glycoprotein in Patients With Endometriosis
Authors: MATHUR SUBBI PILLAI , ZHOU GARY X. , JIANG HAIXIANG , BUTLER WILLIAM J. , ZHANG HUAMEI , ARNAUD PHILIPPE
Published: September 2011

PROBLEM: Identifying the endometrial antigens inciting autoimmunity is important in setting up an antibody assay for a non‐invasive diagnosis and clinical monitoring of endometriosis.

METHODS: Two‐dimensional gel electrophoresis of endometrial extracts, Western blot analysis, passive hemagglutination and enzyme‐linked immunosorbent assay (ELISA), amino acid sequencing and molecular studies were done on chosen antigens. Forty‐six women with endometriosis, 4 women with uterine leiomyomata, 4 with pelvic adhesions, 3 with repeat Cesarean sections (conditions that coexist with or predispose to endometriosis) and 46 controls participated.

RESULTS: Antigens with molecular weights (MW) of 64 kDa [isoelectric point (pI) of 3.5‐4.0] and 72 kDa (pI of 4.5) bound to IgG in all patients with endometriosis, but not the controls. Amino acid sequencing of the proteins revealed that they had homology to alpha 2‐Heremans Schmidt (HS) glycoprotein (MW: 64 kDa) and transferrin (MW: 72 kDa). Endometriosis patients had significant antibody levels to these two proteins (predictive value of 80–90%). The analysis of patients' endometrial RNA detected the message for alpha 2‐HS glycoprotein and transferrin. Albumin (pI 5.5) and collagen (pI 3.5) failed to elicit antibody responses.

CONCLUSIONS: Patients with endometriosis have significant antibodies to endometrial transferrin and alpha 2‐HS‐glycoprotein. We can effectively use an antibody assay using these antigens for diagnosing endometriosis.
Changes in Soluble CD4 and CD8 Proteins in Healthy Pregnant and Postpartum Women
Authors: Watanabe Mikio , Iwatani Yoshinori , Hidaka Yoh , Mitsuda Nobuaki , Amino Nobuyuki
Published: September 2011

PROBLEM: We examined physiological changes in serum levels of soluble CD4 (sCD4) and soluble CD8 (sCD8) during pregnancy and 1–12 months postpartum to study changes in the maternal immune system during and after pregnancy.

METHOD: The serum concentrations of sCD4 and sCD8 were measured by enzyme immunoassay in the sera separated from blood samples withdrawn from healthy women in the 1st, 2nd, and 3rd trimesters of pregnancy and 1,4,7, and 10–12 months postpartum (n=182) and healthy non‐pregnant women (n=25), and in 90 of the women, the changes in sCD4 and sCD8 were compared with changes in the number of peripheral CD4+ and CD8+ cells measured by flow cytometry.

RESULTS: The serum concentration of sCD4 decreased throughout pregnancy, from the first trimester, and recovered gradually after delivery. The serum concentration of sCD8 did not change significantly during or after pregnancy compared to the concentration in the nonpregnant controls, but the concentration 1 month postpartum was significantly higher than that in the 3rd trimester. The numbers of CD4+ and CD8+ cells decreased during pregnancy but did not change significantly after delivery. Interestingly, the ratio of the serum sCD4 level to the number of CD4+ cells decreased and the ratio of the sCD8 level to the number of CD8+ cells increased in the first and second trimesters of pregnancy, but these ratios were within the normal range from the third trimester of pregnancy to 10–12 months postpartum.

CONCLUSIONS: Decreases in serum sCD4 concentration and in the ratio sCD4/CD4+ cells, and an increase in the ratio sCD8/CD8+ cells may be important factors in the immunological changes that occur during pregnancy.
In Vitro Sperm Phagocytosis by Human Peritoneal Macrophages in Endometriosis‐Associated Infertility
Authors: Jha P. , Farooq A. , Agarwal N. , Buckshee K.
Published: September 2011

PROBLEM: The mechanism of infertility in women with endometriosis remains obscure. In this context the role of peritoneal macrophage's sperm phagocytic ability in infertile women with and without endometriosis was investigated.

METHOD: Analysis of peritoneal macrophage concentration and in vitro sperm phagocytosis in 10 infertile women with endometriosis and 10 infertile women without endometriosis was carried out.

RESULTS: It was observed that infertile women with endometriosis are associated with higher concentration and higher sperm phagocytic activity of peritoneal macrophages than those without endometriosis.

CONCLUSION: Despite the small number of women in this study, it may be concluded that the increased number of peritoneal macrophages coupled with enhanced in vitro sperm phagocytosis may disturb fertilization and thereby cause infertility.
Identification of Human Sperm Antigen Recognized by Serum of an Immunoinfertile Woman: A Candidate for Immunocontraception
Authors: Suri A. , Chhabra S. , Upadhyay S.
Published: September 2011

PROBLEM: It has been well documented that antisperm antibodies can be causative factors of infertility. In this study we have identified an antigen on human sperm surface using serum of an immunoinfertile woman; it is thus a candidate for immunocontraception.

METHOD: Thirty‐three women of reproductive age who were infertile were screened for presence of antisperm antibodies by indirect immunofluorescence and agglutination assay. The serum of one such woman, SU‐4, reacted with her husband's as well as normal donor sperm and recognized a band of apparent molecular weight of 71‐kDa on Western blot. Anti‐71 ‐kDa antiserum was raised in rabbit by eluting 71 ‐kDa protein and was characterized by agglutination test, immunofluorescence assay, transmission electron microscopy, flow cytometry, and sperm‐egg interaction in mouse system.

RESULTS: Interestingly, sera raised in rabbit against 71‐kDa antigen, was identified by immunoinfertile serum of SU‐4, revealed similar results of localization of human acrosome. Anti‐71‐kDa antibodies showed cross‐reactivity with other species of sperm, demonstrated inhibition of sperm attachment to oocytes in an in vitro mouse system, and revealed surface binding of human live sperm by flow cytometry. Transmission electron microscopy documented the presence of 71‐kDa antigen in the acrosomal compartment.

CONCLUSION: This study has put in evidence an antigen of apparent molecular weight of 71‐kDa in all donor sperm tested in this study. The presence of this antigen on the sperm of several species will enable us to determine the efficacy of this antigen in controlling fertility in vivo in both rodents and primates. This antigen may be a candidate for immunocontraception.
Human Cytotrophoblastic Cells Absorb the NK Blocking Activity of Monoclonal BA11
Authors: Cadavid A.P. , Guilbert L.J. , Jalali G.R. , Underwood J.L. , Mowbray J.F. , Clark D.A.
Published: September 2011

PROBLEM: R80K is a polymorphic alloantigeneic protein present on human placental trophoblast and on paternal B lymphocytes and monocytes. This protein, unlike the former candidate TLX antigen, stimulates a protective maternal immune response in vivo. A murine monoclonal BA11 antibody, directed against R80K, prevents abortion in three murine pregnancy‐failure models and inhibits human and murine NK activity. We attempted to define the target of BA11 in the human NK assay system.

METHODS: A CELISA method was used to detect R80K antigen on the surface of different cells using the BA11 antibody. The effect, on human peripheral blood NK activity against K562, by BA11 before and after absorption by different cells, including the K562 target, was determined.

RESULTS: R80K was detected on term placental syncytio and cytotrophoblast and on BeWo cells, by CELISA. BA11 suppressed NK lysis of K562 cells in a dose‐dependent manner. Absorption of the BA11 by BeWo and by cytotrophoblastic cells significantly decreased the NK‐inhibitory activity. There was minimal absorption by K562 and BA11‐pretreated K562 cells remained susceptible to NK lysis. By contrast, BA11‐pretreated peripheral blood cells lost all NK activity.

CONCLUSIONS: The inhibition of NK killing of K562 cells by BA11 is more complex than a simple masking of a trophoblast cell‐associated molecule in K562 necessary for recognition in NK cells.
Regulation of Abortion by γλ T Cells
Authors: Arck Petra C. , Ferrick David A. , Steele‐Norwood Darlene , Croitoru Ken , Clark David A.
Published: September 2011

PROBLEM: T cells are present at the feto‐maternal interface, but their function during pregnancy has not been fully elucidated. T cells bearing γλ T‐cell receptor (TCR) may be particularly important, as some subsets can react to trophoblast cells by producing cytokines, such as interleukin‐2 (IL‐2).

METHOD: We depleted T cells bearing the γλ receptor by injecting monoclonal antibodies (mAB) into females of the abortion‐prone animal model CBA x DBA/2. We investigated the percentage and number of γλ T‐cell receptor positive (TCR)+ cells in decidua and spleen during pregnancy in control and γλ‐depleted female mice. Pregnant females were also exposed to ultrasonic sound stress to boost the abortion rate.

RESULTS: Stress failed to increase the abortion rate in the γλ TCR‐depleted mice. FACScan analysis show that the ratio of cells bearing the γλ TCR dramatically decreased after injection of mAB to the γλ TCR in spleen and decidua, these cells recovered six days after depletion, showing a change in cytokine pattern. Levels of TNF‐α in decidual γλ T cells decreased; similar effects of decreasing Th1 cytokines could be observed in splenic γλ T cells. We further identified increased levels of intracellular TNF‐α in the Vλ4 subset in the decidua, compared to spleen.

CONCLUSIONS: Trophoblast recognition by the Vλ4 T‐cell subset in the decidua may cause the release of abortogenic cytokines such as TNF‐α. Depletion of such γλ TCR T cells during early pregnancy may promote successful pregnancy outcome in normal pregnancy and prevent stress‐induced abortions.
Increased Expression of Cell Surface Markers on Endometrial γδ T‐Cell Receptor+ Intraepithelial Lymphocytes Induced by the Local Presence of the Sheep Conceptus
Authors: Liu Wen‐Jun , Gottshall Susan L. , Hansen Peter J.
Published: September 2011

PROBLEM: In sheep, γδ T‐cell receptor (TCR)+ cells are a major lymphocyte subpopulation in the luminal epithelium of the uterine endometrium. During late pregnancy, this population of T cells increases in number and becomes more granulated. This study was performed to determine whether this apparent activation was induced by local effects of the conceptus or systemic effects of pregnancy.

METHODS: The unilaterally‐pregnant ewe, in which the conceptus is surgically confined to one uterine horn, was used to distinguish between systemic and local effects of pregnancy on function of endometrial γδ TCR+ intraepithelial lymphocytes. Lymphocytes collected from peripheral blood, and from the endometrial luminal epithelium of cyclic and unilaterally‐pregnant ewes (day 140 of gestation) were analyzed by flow cytometry.

RESULTS: As compared to γδ TCR+ lymphocytes in peripheral blood, γδ TCR+ intraepithelial lymphocytes from non‐pregnant ewes had a lower percentage of cells staining positive for CD25, CD44, and L‐selectin. There was no effect of pregnancy on the percentage of γδ TCR+ peripheral blood lymphocytes staining positive for CD25, CD44, CD29, or L‐selectin. Similarly, the percentage of intraepithelial lymphocytes staining positive for these antigens was similar for cells collected from cyclic ewes and cells from the nonpregnant uterine horn of unilaterally‐pregnant ewes. In contrast, there was a higher percentage of CD25, CD44, CD29, and L‐selectin+ cells for γδ TCR+ intraepithelial lymphocytes from the conceptus‐containing uterine horn of pregnant ewes than for γδ TCR+ intraepithelial lymphocytes from the endometrium of cyclic ewes or from the nonpregnant uterine horn of pregnant ewes.

CONCLUSION: The local presence of the conceptus causes changes in cell surface marker expression on endometrial γδ TCR+ intraepithelial lymphocytes during pregnancy. These changes may reflect conceptus‐induced activation of this population of cells.
Differential Expression of IGF‐I and IGF‐II in Eutopic and Ectopic Endometria of Women With Endometriosis and in Women Without Endometriosis
Authors: SBRACIA M. , SCARPELLINI F. , ZUPI E. , MANNA C. , MARCONI D. , ROMANINI C. , ALO P. , DI TONDO U. , GRASSO J.A.
Published: September 2011

PROBLEM: Factors regulating the development, growth, and differentiation of endometrial cells of endometriotic lesions are poorly understood. To investigate the paracrine‐autocrine regulation of ectopic endometrial cell growth, the expression of IGF‐I and IGF‐II were studied.

METHOD: Tissue specimens of eutopic and ectopic endometria were obtained from eight patients with endometriosis at laparoscopy and from the endometria of 14 women without endometriosis as controls. They were tested for the expression of IGF‐I and IGF‐II by immunohistochemical analysis.

RESULTS: Immunohistochemical study for IGF‐I in controls showed a more intense staining during the proliferative phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis a reduction in the staining was observed, whereas in epithelial cells of fibrotic peritoneal adhesions an intense immunostaining for IGF‐I was observed. Immunohistochemical study of IGF‐II in controls showed a more intense staining during secretory phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis, a reduction in the staining was observed, whereas in epithelial cells of fibrotic peritoneal adhesions an intense immunostaining for IGF‐I was observed. Immunohistochemical study of IGF‐II in controls showed a more intense staining during secretory phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis, a reduction in the staining was observed, whereas in epithelial cells of ovarian lesions and fibrotic peritoneal adhesions, no immunostaining for IGF‐II was observed.

CONCLUSIONS: In endometriosis there is an alteration of mechanisms regulating cell proliferation and differentiation.
Growth‐Regulated α Expression in Human Preovulatory Follicles and Ovarian Cells
Authors: Oral Engin , Seli Emre , Bahtiyar Mert O. , Jones Ervin E. , Arici Aydin
Published: September 2011

PROBLEM: Around the time of ovulation the number of neutrophils increases in the theca of the leading follicle. We hypothesized that growth‐regulated a (GROα), a neutrophil chemoattractant/activating factor, may be a modulator of periovulatory neutrophil chemotaxis.

METHOD: GROα levels were measured in follicular fluids (n = 61). Granulosa‐lutein and ovarian stromal cells were also cultured. After experimental paradigms, GROα mRNA was evaluated by Northern analysis, GROα in follicular fluids, and culture supernatants were quantified using ELISA.

RESULTS: In follicular fluids the mean pre‐human chorionic gonadotropin (hCG) GROα level was 51 ± 24 (±SEM) pg/ml, post‐hCG it was 210 ± 20 pg/ml (P = 0.04). GROα was produced constitutively by ovarian stromal and granulosa‐lutein cells. Interleukin‐α (IL‐1α) and tumor necrosis factor‐α (TNF‐α) further stimulated GROα production. Treatment of stromal cells with hCG also stimulated GROα production.

CONCLUSION: GROα is a constituent of periovulatory follicular fluid. Ovarian stromal and granulosa‐lutein cells express the GROα mRNA and produce the protein. The regulation of GROα by cytokines and hCG suggests that GROα may play a role in the process of ovulation.
Anti‐Endometrial Autoantibodies in Women with a Diagnosis of Infertility
Authors: Palacio J.R. , Iborra A. , Martinez P. , Gris J.M. , Andolz P.
Published: September 2011

PROBLEM: The purpose of this study was to investigate the frecuency of anti‐endometrial antibodies (AEA) in infertile women.

METHOD OF STUDY: Sera from fertile women (n = 6), and from patients with ovulatory dysfunction (n = 11), tubal obstruction (n = 9) and unexplained infertility (n = 5) were investigated for the presence of anti‐endometrial membrane antibodies. We used two human endometrial cancer cell lines and human endometrial cells from gynecological biopsies as an antigenic source for analysis. The immunoenzymatic assay (ELISA) was performed with cultured endometrial cells in monolayers. Immunoblot analysis was performed with these two cell lines.

RESULTS: A good correlation between the response with each cell line and with human endometrial cells was obtained, indicating that the antigens analyzed were probably similar. Endometrial antibodies were detectable in a high percentage of women with tubal obstruction (77.8 and 66.7%, respectively) and ovulatory dysfunction (54.5 and 45.5%, respectively). Unexplained infertility showed anti‐endometrial immunological response (40 and 60%, respectively). Some endometrial antigens in infertile women are the target for autoimmune response. The serum from a patient with tubal obstruction and ovulatory dysfunction showed two antigens by immunoblot, with molecular weights of 97 and 50 kDa.

CONCLUSION: The presence of anti‐endometrial antibodies, detected by ELISA, is associated with infertility, mainly with ovulatory dysfunction and tubal obstruction. Some endometrial antigens may be involved in these two pathologies.
HLA Class I Expression on the Materno‐Fetal Interface
Authors: Hammer Astrid , Hutter Heinz , Dohr Gottfried
Published: September 2011

PROBLEM: The conditions that permit the genetically dissimilar (haplo‐non‐identical) human fetus to evade rejection by its mothers immune system have been the subject of intense interest for several years. As the placental cells, which are in contact with maternal blood or tissue, are devoid of HLA class II antigens, the interest has focused on the expression of HLA class I molecules.

METHOD OF STUDY: Recent findings on the constitutive, transcriptional, and translational expression of HLA class I molecules on anatomically and morphologically different sub‐populations of trophoblast cells will form the basis of this article.

RESULTS: The expression of HLA class I molecules in the trophoblast cells, forming the materno‐fetal junctional zone is inhomogeneous. It differs depending on the differentiation and location of trophoblast cells within the placenta and furthermore on the stage of gestation. On the transcriptional level HLA‐A, ‐B, ‐C, ‐E, and ‐G could be detected on individual trophoblast populations, whereas only HLA‐C and HLA‐G seem to be translated to protein. CONCLUSIONS: The expression of HLA class I antigens by trophoblast cells is not simply suppressed. Instead, less polymorphic HLA‐G and HLA‐C antigens are carefully selected from the class I multigene family. This gives rise to the assumption that these two HLA class I molecules play an important role in the maintenance of pregnancy.