bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro antibacterial effect of calcium hydroxide combined with chlorhexidine or iodine potassium iodide on Enterococcus faecalis
Authors: Sirén Eva K. , Haapasalo Markus P. P. , Waltimo Tuomas M. T. , Ørstavik Dag
Published: July 2004

Several studies have shown a higher success rate of root canal therapy when the canal is free from bacteria at the time of obturation. Treatment strategies that are designed to eliminate this microflora should include agents that can effectively disinfect the root canal. Enterococcus faecalis is often associated with persistent endodontic infections. While in vivo studies have indicated calcium hydroxide to be the most effective all‐purpose intracanal medicament, iodine potassium iodide (IKI) and chlorhexidine (CHX) may be able to kill calcium hydroxide‐resistant bacteria. Supplementing the antibacterial activity of calcium hydroxide with IPI or CHX preparations was studied in bovine dentine blocks. While calcium hydroxide was unable to kill E. faecalis in the dentine, calcium hydroxide combined with IKI or CHX effectively disinfected the dentine. The addition of CHX or IKI did not affect the alkalinity of the calcium hydroxide suspensions. It may be assumed that combinations also have the potential to be used as long‐term medication. Cytotoxicity tests using the neutral red method indicated that the combinations were no more toxic than their pure components.
Mechanism of Fas‐mediated cell death and its enhancement by TNF‐α in human salivary gland adenocarcinoma cell line HSG
Authors: Chosa Naoyuki , Kyakumoto Seiko , Kito Noriko , Kamo Masaharu , Sato Nobuko
Published: July 2004

Fas‐mediated cell death in a human salivary gland adenocarcinoma cell line (HSG) was induced by treatment of the cells with agonistic anti‐Fas antibody (CH‐11), and this cell death was enhanced by pretreatment with tumor necrosis factor alpha (TNF‐α). The mode of cell death was apoptosis, because it was accompanied by caspase activation and the cleavage of poly(ADP‐ribose) polymerase. The TNF‐α treatment of the cells increased the expression of Fas, which was accompanied by the activation of nuclear factor κB (NFκB). These results suggest that the enhancement of the apoptosis caused by TNF‐α resulted from increased sensitivity of the HSG cells to CH‐11‐mediated apoptosis due to induction of Fas protein by TNF‐α via the activation of NFκB. In order to elucidate the apoptosis signaling pathway, we examined the effect of various caspase inhibitors on the apoptosis induced by CH‐11. Fas‐mediated apoptosis of HSG cells was slightly inhibited by the caspase‐9 inhibitor although it was mainly inhibited by that for caspase‐8. Based on this finding, we consider CH‐11‐induced apoptosis in HSG cells to be mainly mediated by the type I death signaling pathway that is caused by a caspase cascade initiated by the activation of caspase‐8 at the death‐inducing signaling complex (DISC).
The adhesion between fiber posts and root canal walls: comparison between microtensile and push‐out bond strength measurements
Authors: Goracci Cecilia , Tavares Andrea Urbano , Fabianelli Andrea , Monticelli Francesca , Raffaelli Ornella , Cardoso Paulo Capel , Tay Franklin , Ferrari Marco
Published: July 2004

Aim of the study was to compare the trimming and non‐trimming variants of the microtensile technique with the ‘micro’ push‐out test in the ability to measure accurately the bond strength of fiber posts luted inside root canals. In 15 endodontically treated teeth (Group A), fiber posts were cemented with Excite DSC in combination with Variolink II (Ivoclar‐Vivadent). In 15 roots RelyX Unicem (3M‐ESPE) was used for fiber post luting (Group B). Within each group, the bond strength of cemented fiber posts was assessed with the trimming and non‐trimming microtensile technique, as well as with the push‐out test. The great number of premature failures (16.9% in Group A, 27.5% in Group B) and the finding of high standard deviation values make questionable the reliability of the trimming microtensile technique. With the non‐trimming microtensile technique, only five sticks were obtained from a total of six roots. The remaining specimens failed prematurely during the cutting phase. With the push‐out test no premature failure occurred, the variability of the data distribution was acceptable, and regional differences in bond strength among root levels could be assessed. Relatively low values of bond strength were, in general, recorded for luted fiber posts. In conclusion, when measuring the bond strength of luted fiber posts, the push‐out test appears to be more dependable than the microtensile technique.
Bone remodeling during prenatal morphogenesis of the human mental foramen
Authors: Radlanski Ralf J. , Renz Herbert , Lajvardi Souzan , Schneider Richard A.
Published: July 2004

From a morphogenetic point of view, the mental foramen of the mandible is a highly suitable model to study the interactions of different tissues such as nerves, vessels, mesenchymal cells, cartilage, and bone. In previous work, we provided a three‐dimensional description of the mental foramen at different developmental stages, and now we complement those studies with a three‐dimensional visualization of different bone remodeling activities around the mental foramen. Histological serial sections of human embryos and fetuses, ranging in size from 25 to 117 mm crown‐rump‐length (CRL), were used to characterize the bone remodeling activity (apposition, inactivity, and resorption). We quantified and reconstructed this activity in three dimensions, and included information on the spatial relationship of the nerves, vessels, and dental primordia. In general, the mandible showed strong apposition at its outer surfaces. The brim of the mental foramen, however, displayed changing remodeling activity at different stages. In the depth of the bony gutter, which provides space for the nerve and the blood vessels, we found bone resorption beneath the inferior alveolar vein. Bone was also resorbed in proximity to the dental primordia. In future studies, we will relate gene expression data to these morphological findings in order to identify molecular mechanisms that regulate this complex system.
Effects of a fluoride etchant on resin bonding to titanium‐aluminum‐niobium alloy
Authors: Yanagida Hiroaki , Taira Yohsuke , Atsuta Mitsuru
Published: July 2004

This investigation was carried out in order to evaluate ammonium hydrogen fluoride (AHF) and cupric chloride (CC) as components of a metal etchant. The surface of cast titanium‐aluminum‐niobium (Ti‐6Al‐7Nb) was air‐abraded with alumina, etched for 10 s, and rinsed with water. A phosphate or a thiophosphate primer was applied to the bonding area, and an acrylic rod was bonded to the specimen with a tri‐n‐butylborane‐initiated self‐curing luting agent. Shear bond strengths were determined after thermocycling (4°C and 60°C) for 10,000 cycles. The average bond strength was significantly influenced by thermocycling, AHF, and primer, but was not influenced by CC. The maximum average bond strengths were obtained when the etchant consisted of 5mass% AHF, with and without 0.3mass% CC. Microphotographs showed that numerous micropits were created on the etched surface, suggesting increased micromechanical retention. In conclusion, chemical etching with 5mass% AHF significantly improved the durability of resin bonding to Ti‐6Al‐7Nb.
Evidence of chemisorption of maleic acid to enamel and hydroxyapatite
Authors: Fu Baiping , Yuan Jun , Qian Weixin , Shen Qingyi , Sun Xuemei , Hannig Matthias
Published: July 2004

Maleic acid has been used as an etchant or non‐rinse conditioner in adhesive dentistry. However, the inherent mechanisms of the interaction of maleic acid with hydroxyapatite/enamel have never been fully elucidated. The purpose of this study was to provide evidence for the chemisorption of maleic acid onto hydroxyapatite/enamel, and to identify the reaction products obtained following the interaction of maleic acid with hydroxyapatite. Hydroxyapatite particles were dissolved in a 15% (w/v) aqueous solution of maleic acid (pH = 0.98). Half of the solution was dried to obtain a desiccated mixture. This mixture, hydroxyapatite, maleic acid and self‐prepared calcium maleate were analysed by X‐ray diffraction (XRD). Acetone was added to the other half of the solution to obtain a precipitate. This precipitate, hydroxyapatite, maleic acid, unetched enamel and maleic acid‐etched enamel were analysed by X‐ray photoelectron spectroscopy (XPS). The precipitate was also analysed by 1H NMR. A new binding energy, indicating carboxylate groups, was detected by XPS on the precipitate and maleic acid‐etched enamel surface. XRD data indicated the formation of calcium maleate and calcium hydrogen phosphate after the reaction. NMR data revealed that one carboxylic group of maleic acid reacted with hydroxyapatite. Hence, maleic acid can chemisorb to hydroxyapatite and enamel via ionic interactions.
Risk Factors and Mechanisms of Preterm Delivery in Malawi
Authors: Abrams Elizabeth T. , Milner Danny A. , Kwiek Jesse , Mwapasa Victor , Kamwendo Deborah D. , Zeng Donglin , Tadesse Eyob , Lema Valentino M. , Molyneux Malcolm E. , Rogerson Stephen J. , Meshnick Steven R.
Published: July 2004

Problem:  We examined risk factors and mechanisms of preterm delivery (PTD) in malaria‐exposed pregnant women in Blantyre, Malawi.

Method of study:  The human immunodeficiency virus (HIV), malaria, syphilis, and anemia were assessed in a cross‐sectional study of 572 pregnant women. In a nested case–control study, chorioamnionitis (CAM) was examined; tumor necrosis factor (TNF)‐α, interleukin (IL)‐6, IL‐8, macrophage inflammatory protein (MIP)‐1α, monocyte chemotactic protein (MCP)‐1, transforming growth factor (TGF)‐β, cortisol, and corticotropin‐releasing hormone were measured in placental, maternal and/or cord blood.

Results:  HIV, infrequent antenatal clinic attendance, low‐maternal weight, no intermittent preventive malaria therapy (IPT), and CAM were associated with PTD, while malaria was not. Of the 18 compartmental cytokine measurements, elevations in placental and/or cord IL‐6 and IL‐8 were associated with both CAM and PTD. In contrast, there was no overlap between the cytokines affected by malaria and those associated with PTD.

Conclusions:  The HIV and CAM were the major infections associated with PTD in this study. CAM, but not malaria, causes PTD via its effect on proinflammatory cytokines.
Molecular Cloning, Expression of Testicular Transcript Abundant in Germ Cells and Immunobiological Effects of the Recombinant Protein
Authors: Verma Saguna , Mohapatra Bhagyalaxmi , Jagadish Nirmala , Selvi Ramasamy , Roy Pragyan , Rana Ritu , Lakshmi Kavitha , Suri Anil
Published: July 2004

Problem:  It has been well documented that antisperm antibodies can be causative factors for infertility. In this report we have identified a protein on human sperm referred as human sperm‐associated protein (HSAP) using serum of an immunoinfertile woman; it is thus a sperm‐specific protein – a candidate molecule for control of fertility.

Method of study:  An immunoinfertile woman serum showing head–head sperm agglutination and acrosomal localization, reacted with human sperm protein of apparent molecular weight of 48 kDa on Western blot. Anti‐48 kDa antiserum was raised in rabbit by eluting 48 kDa protein and was used to screen the human testis cDNA expression library. A putative positive hsap cDNA clone was obtained, sequenced and subjected to tissue specificities studies by Northern blotting. The cell type‐specific expression was done using in situ RNA hybridization studies. To obtain recombinant HSAP (r‐HSAP), hsap cDNA was cloned in pET 22b(+) expression vector. r‐HSAP was expressed as polyhistidine fusion protein in Escherichia coli and purified. Rabbits were immunized with the purified r‐HSAP, which led to generation of antibodies. In order to evaluate in vitro immunocontraceptive potential, the anti‐r‐HSAP antibodies were characterized by agglutination assay, zona‐free hamster egg penetration assay, indirect immunofluorescence (IIF) assay, and by flow cytometry analysis.

Results:  We have cloned a human testis gene encoding a protein (HSAP) of 328 amino acids. Antibodies against the purified recombinant protein specifically recognized approximately 40 kDa r‐HSAP, and a cognate 48 kDa protein band in human sperm extract in Western blot procedure. The anti‐r‐HSAP antibodies localized acrosomal compartment, inhibited sperm binding/attachment in zona‐free hamster penetration assay and revealed surface binding with human live sperm by flow cytometry. The cDNA sequence has been submitted to EMBL and has been given the accession number Y16676.

Conclusion:  This study has put in evidence that novel sperm‐specific r‐HSAP has role in sperm function and may have application in the development of a contraceptive vaccine. The availability of the recombinant protein will facilitate studies on the assessment of its potential as a contraceptive immunogen.
Structural and Functional Heterogeneity in the CD200R Family of Immunoregulatory Molecules and their Expression at the Feto‐maternal Interface
Authors: Gorczynski Reginald M. , Chen Zhiqi , Clark David A. , Kai Yu , Lee Lydia , Nachman Joseph , Wong Simon , Marsden Philip
Published: July 2004

Problem:  We have shown that CD200Fc, a chimeric molecule including the extracellular domain of CD200 and a murine immunoglobulin (Ig)G2a Fc region, regulates immune responses and prevents T helper (Th)1 cytokine‐triggered spontaneous abortions in mice. CD200 is expressed on a subpopulation of uterine decidua cells and on trophoblast, both in the mouse and human. The receptor(s) for CD200, CD200R(s), was not previously well‐characterized.

Methods:  5′‐rapid amplification of cDNA ends (RACE), cDNA and genomic DNA clone analysis were used to identify a family of CD200Rs on mouse chromosome 16, juxtaposed to the CD200 gene, named CD200R1, R2, R3, and R4. Northern blot and reverse transcriptase polymerase chain reaction (RT‐PCR) analysis was used to detect expression of different CD200R subtypes in different organs. Rabbit polyclonal and rat monoclonal antibodies (mAbs) to CD200R isoforms was used for fluoresence‐activated cell sorter (FACS) analysis, to test for immunomodulatory effects on allogeneic mixed‐lymphocyte responses in vitro, and for immunohistochemistry.

Results:  The CD200Fc was able to interact physically with each of the CD200Rs expressed on the cell surface. Northern blot and RT‐PCR analyses indicated distinct patterns of CD200R isoform mRNA expression in different tissues and FACS analyses confirmed unique cell‐ and tissue‐specific expression of the different CD200Rs. mAbs directed against the different isoforms modified the development of in vitro alloimmune responses. The addition of anti‐CD200R1/R4 elicited immunomodulatory responses in vitro comparable to findings with CD200Fc, but different from the effects of anti‐CD200R2–3.

Conclusions:  These data provide evidence for a family of CD200R molecules in the mouse genome and defines the existence of previously unrecognized diversity in the CD200/CD200R immunomodulatory gene member family. Although this gene member family is clustered in the genome, the different CD200Rs and CD200 exhibit distinct expression patterns and functional properties. Restricted CD200R isoform expression at the feto‐maternal interface suggests CD200:CD200R interactions may serve important function(s) determining the successful outcome of pregnancy.
Interleukin‐4 Gene Polymorphism is not Involved in the Risk of Recurrent Pregnancy Loss
Authors: Saijo Yasuaki , Sata Fumihiro , Yamada Hideto , Konodo Tomoko , Kato Emi Hirayama , Kataoka Soromon , Shimada Shigeki , Morikawa Mamoru , Minakami Hisanori , Kishi Reiko
Published: July 2004

Problem:  Enhanced secretion of type‐2 T‐helper (Th2) cytokine is a characteristic feature in normal physiological pregnancy. A study has demonstrated defective production of interleukine‐4 (IL‐4) and other Th2 cytokine in women with recurrent pregnancy loss (RPL). Several studies have suggested that IL‐4 variable number of tandem repeat (VNTR) gene polymorphism is probably associated with different IL‐4 production.

Methods of study:  The IL‐4 VNTR genotypes were assessed in 109 Japanese women with RPL and 210 ethnically matched women experiencing at least one live birth and no spontaneous abortion.

Results:  No significant differences in IL‐4 VNTR genotype frequencies were found between the RPL and the control [B1B1 genotype (reference); B1/B2 and B2/B2 genotypes, odds ratio, 0.91; 95% confidence interval, 0.58–1.45].

Conclusion:  The present study suggests that the IL‐4 VNTR allele is not a major genetic regulator in RPL.
Serum P‐selectin Level During Controlled Ovarian Hyperstimulation – a Preliminary Report
Authors: Orvieto Raoul , Badir Walid , Bar Jacob , La Marca Antonio , Ashkenazi Jacob , Avrech Ori M. , Fisch Benjamin
Published: July 2004

Objective:  To measure levels of serum P‐selectin in patients undergoing controlled ovarian hyperstimulation (COH) cycles and to determine their possible correlation to COH variables.

Setting:  Large university‐based infertility and in vitro fertilization unit.

Patients:  Fourteen consecutive patients undergoing our routine COH protocol for unexplained infertility.

Interventions and Main Outcome Measures:  Blood was drawn three times during the COH cycle: (1) day 2 or 3 of the menstrual cycle, before gonadotropin treatment (Day‐0); (2) day of or prior to human chorionic gonadotropin (hCG) administration (Day‐hCG); and (3) day of ovulation (Day‐OVU). Serum levels of sex steroids and P‐selectin were compared among the three time points. P‐selectin was measured with a commercial quantitative sandwich immunoassay technique. To reduce interpatient variability, the percent difference between the Day‐0 (non‐stimulated, basal) level and the Day‐hCG and Day‐OVU levels was calculated.

Results:  P‐selectin level on Day‐hCG was significantly higher than on Day‐0 (P < 0.05) and non‐significantly higher than on Day‐OVU (P < 0.12). No significant correlations were observed between serum P‐selectin and patient age, amount of gonadotropins used, or estradiol or progesterone level.

Conclusion:  The increase in serum P‐selectin level during COH until peak estradiol suggests that COH may potentiate a state of platelet activation which is substantially attenuated after hCG administration.
11β‐Hydroxysteroid Dehydrogenase Inhibitor Carbenoxolone Stimulates Chorionic Gonadotropin Secretion from Human Term Cytotrophoblast Cells Differentiated In Vitro*
Authors: Nacharaju Vijaya L. , Divald Andras , McCalla Cyrus O. , Yang Lianfu , Muneyyirci‐Delale Ozgul
Published: July 2004

Problem:  To investigate the effect of altering local glucocorticoid concentration on human chorionic gonadotropin (hCG) production by cultured placental trophoblast cells.

Method of study:  Human placental trophoblasts were isolated from fresh placentas. Cytotrophoblasts were purified and placed into 24‐well multiplates. For cultivation Dulbecco's modified Eagle's medium (DMEM) with 15 mm HEPES and 15% FBS was used. 11β‐Hydroxysteroid dehydrogenase (11β‐HSD) activity and its inhibition by carbenoxolone (CE) were measured in cultured cells. Cultures were exposed to CE for 16–20 hr. Overnight production of hCG was measured by radioimmunoassay in control and treated cells.

Results:  The 11β‐HSD activity in these cultures was inhibited by nm concentrations of CE, the apparent Ki being 2.5 nm. Inhibition of 11β‐HSD activity with 0.1 nm CE resulted in 1.5‐fold increase in the production of hCG.

Conclusions:  Increasing local glucocorticoid concentration by the inhibition of 11β‐HSD results in higher hCG secretion, which in turn enhance cell differentiation.
Life‐Threatening Bleeding, Pregnancy and Lupus Anticoagulant: Success after Steroid and Anticoagulant Therapy
Authors: Alijotas‐Reig Jaume , Ferrer‐Raventos Juan Carlos
Published: July 2004

Problem:  The clinical manifestations associated with the presence of the lupus anticoagulant (LAC) are usually thrombotic and/or obstetric complications, specially miscarriages and/or repeated fetal deaths. The bleeding episodes in patients with LAC with no other coagulation disorders are exceptional, especially during pregnancy, intra‐ and postpartum.

Method of study:  Here we present two cases of patients with classification criteria of primary antiphospholipid antibody syndrome, a history of recurrent complicated pregnancies and massive bleeding during abortion and postpartum. We studied anticardiolipin (IgG/IgM), beta2glicoprotein‐1 (IgG) and antimitochondrial (type 5) antibodies, LAC, rapid plasma reagin, coagulation test, clotting factors and, placental vascular flow from the second trimester of their last pregnancies.

Results:  LAC was repeatedly detected in both cases. The prothrombin time and clotting factors including factors II and XIII were normal. Intrauterine growth restriction was also observed in both patients. Pre‐eclampsia was detected in patient 2. We tried enoxaparin (60–80 mg/day) and prednisone (1 mg/kg/day) therapy and no maternal, hemorrhagic or thrombotic episodes were observed.

Conclusions:  Exceptionally, LAC may be associated with recurrent, life‐threatening bleeding, even in patients with no prothrombin deficiency. The close follow‐up and early anticoagulant and steroid therapy seems to be able to control thrombotic and hemorrhagic complications during the pregnancy and postpartum.
The Effect of Inflammatory Cytokines on Secretion of Macrophage Colony‐Stimulating Factor and Monocyte Chemoattractant Protein‐1 in Human Granulosa Cells
Authors: Kawano Yasushi , Fukuda Junichiro , Itoh Hiroko , Takai Noriyuki , Nasu Kaei , Miyakawa Isao
Published: July 2004

Problem:  In order to investigate the role of macrophage colony‐stimulating factor (M‐CSF) and monocyte chemoattractant protein ‐1 (MCP‐1) in human ovulation, we studied the regulation of M‐CSF and MCP‐1 in cultured human granulosa cells.

Method of study:  Immortalized granulosa cells (GC1a) were cultured in serum‐free medium, and incubated with interleukin (IL)‐1α, IL‐1 receptor antagonist (ra) and tumor necrosis factor (TNF)‐α. The supernatants were collected, and M‐CSF and MCP‐1 were measured by ELISA.

Results:  The levels of M‐CSF and MCP‐1 were increased after treatment with IL‐1α (1 nm) and TNF‐α (1 nm) in a time‐dependent manner. The levels of M‐CSF and MCP‐1 were significantly increased after treatment with IL‐1α and TNF‐α in a dose‐dependent manner. However, the levels of M‐CSF and MCP‐1 were significantly decreased by treatment with IL‐1α (1 nm) and/or increasing concentrations of IL‐1 ra.

Conclusions:  Our data indicated that M‐CSF and MCP‐1 were regulated by IL‐1α and TNF‐α. It was suggested that M‐CSF and MCP‐1 may play an important role in human preovulatory processes.
Effect of Hypoxia on Urocortin Production in Human Gestational Trophoblasts In Vitro
Authors: Choy Mei Y. , Leung Tse N. , Leung Po S. , Lau Tze K.
Published: July 2004

Problem:  Urocortin is produced by the placenta throughout pregnancy but its regulation remains unknown. The effect of hypoxia on placental urocortin production is not known. The aim of this study was to determine the effect of in vitro hypoxia on human trophoblastic urocortin production.

Method of study:  Placental explants and primary cultures were incubated in anaerobe hypoxic bags for 24 h in a humidified incubator. Urocortin peptide secretion and mRNA (messenger RNA) production was determined by enzyme‐linked immunosorbent assay and reverse transcription‐polymerase chain reaction, respectively. Morphological and functional integrity was verified by immunohistochemical analysis of urocortin expression. Vascular endothelial growth factor expression was used to verify the generation of cellular hypoxia in our in vitro system.

Results:  Hypoxia did not affect urocortin secretion or mRNA expression in explant and single‐cell cultures. Production was greater from first trimester than term explants and from single‐cell primary cultures more than from explant cultures.

Conclusions:  Hypoxia does not influence human placental urocortin secretion or mRNA expression in vitro.
Endometrial and Peritoneal Macrophages: Expression of Activation and Adhesion Molecules
Authors: Eidukaite Audrone , Tamosiunas Vytas
Published: July 2004

Problem:  Macrophages are highly individualized in tissues and their activities are a reflection of systemic and local environmental signals. The expression of activation (CD69, CD71) and adhesion (CD54) molecules on the surface of CD14+ endometrial macrophages at various phases of the menstrual cycle was compared with the cell surface receptors of peritoneal fluid macrophages.

Method of study:  Two‐colour‐flow cytometry was used to determine the peritoneal and endometrial macrophage phenotype.

Results:  Endometrium macrophages expressed a lower level of CD69+ and CD54+ macrophages than peritoneal macrophages. However, CD71 receptors displayed similar expression in both macrophage populations, endometrium and peritoneal, except during the proliferative phase.

Conclusion:  These findings demonstrate the differences between macrophages from endometrium and peritoneal fluid with regard to CD69, CD71 and CD54 expression. In addition, increased numbers of endometrial macrophages in the late secretory phase of the menstrual cycle suggest that they may play a role in menstruation.
Relationship between Cervical Mucus Interleukin‐8 Concentrations and Vaginal Bacteria in Pregnancy
Authors: Sakai Masatoshi , Ishiyama Akihiko , Tabata Mika , Sasaki Yasushi , Yoneda Satoshi , Shiozaki Arihiro , Saito Shigeru
Published: July 2004

Problem:  High interleukin (IL)‐8 concentration in cervical mucus in the second trimester is a risk factor for premature birth. We investigated the relationship between vaginal pathogens and IL‐8 in cervical mucus.

Method of study:  In 501 women with single pregnancy, vaginal secretions were cultured for bacteria and cervical mucus IL‐8 concentrations were measured between 20 and 24 gestational weeks.

Results:  Lactobacillus species were detected in 56.0% of 84 subjects with high IL‐8 (≥377 ng/mL), significantly less often than in 417 subjects with IL‐8 below 377 ng/mL (84.7%; P < 0.0001). Anaerobic pathogens were detected in 83.3% of high IL‐8 subjects, significantly more often than in normal IL‐8 subjects (43.9%; P < 0.0001). By multivariate analysis, cervical IL‐8 was significantly high only in subjects without Lactobacillus species; they showed a significantly higher prematurity rate than Lactobacillus‐positive subjects.

Conclusions:  Absence of vaginal Lactobacilli was associated with increased cervical IL‐8 and increased risk of premature delivery.
Leukemia Inhibitory Factor: An Important Regulator of Endometrial Function
Authors: Kondera‐Anasz Zdzisława , Sikora Justyna , Mielczarek‐Palacz Aleksandra
Published: July 2004

Problem:  Leukemia inhibitory factor (LIF) is multifunctional cytokine that displays biological activities in different cells, including endometrial cells. The aim of this study is to describe implications of LIF on a physiological function of endometrium.

Method of study:  The role of LIF in the endometrial function is reviewed and summarized from the available literature.

Results:  LIF plays an important role in a physiological function of endometrium. In human endometrial LIF expression depends on cellular localizations, steroid hormones, menstrual stages and a local cytokine network. Stronger LIF expression exists in an endometrial epithelium during a luteal phase of the menstrual cycle, which coincides with the time of an implantation. The impairments of the endometrial LIF expression may play a significant role in the pathological processes involving implantation and the infertility.

Conclusions:  There is a substantial evidence that LIF is a potential regulator of the endometrial function and might be one of the factors that play a key role in human reproduction.
HLA and NK Cell Inhibitory Receptor Genes in Resolving Hepatitis C Virus Infection
Authors: Khakoo Salim I. , Thio Chloe L. , Martin Maureen P. , Brooks Collin R. , Gao Xiaojiang , Astemborski Jacquie , Cheng Jie , Goedert James J. , Vlahov David , Hilgartner Margaret , Cox Steven , Little Ann-Margeret , Alexander Graeme J. , Cramp Matthew E. , O'Brien Stephen J. , Rosenberg William M. C. , Thomas David L. , Carrington Mary
Published: August 2004

Natural killer (NK) cells provide a central defense against viral infection by using inhibitory and activation receptors for major histocompatibility complex class I molecules as a means of controlling their activity. We show that genes encoding the inhibitory NK cell receptor KIR2DL3 and its human leukocyte antigen C group1 (HLA-C1) ligand directly influence resolution of hepatitis C virus (HCV) infection. This effect was observed in Caucasians and African Americans with expected low infectious doses of HCV but not in those with high-dose exposure, in whom the innate immune response is likely overwhelmed. The data strongly suggest that inhibitory NK cell interactions are important in determining antiviral immunity and that diminished inhibitory responses confer protection against HCV.
Localization and Physical Properties Experiments Conducted by Spirit at Gusev Crater
Authors: Arvidson R. E. , Anderson R. C. , Bartlett P. , Bell J. F. , Blaney D. , Christensen P. R. , Chu P. , Crumpler L. , Davis K. , Ehlmann B. L. , Fergason R. , Golombek M. P. , Gorevan S. , Grant J. A. , Greeley R. , Guinness E. A. , Haldemann A. F. C. , Herkenhoff K. , Johnson J. , Landis G. , Li R. , Lindemann R. , McSween H. , Ming D. W. , Myrick T. , Richter L. , Seelos F. P. , Squyres S. W. , Sullivan R. J. , Wang A. , Wilson J.
Published: August 2004

The precise location and relative elevation of Spirit during its traverses from the Columbia Memorial station to Bonneville crater were determined with bundle-adjusted retrievals from rover wheel turns, suspension and tilt angles, and overlapping images. Physical properties experiments show a decrease of 0.2% per Mars solar day in solar cell output resulting from deposition of airborne dust, cohesive soil-like deposits in plains and hollows, bright and dark rock coatings, and relatively weak volcanic rocks of basaltic composition. Volcanic, impact, aeolian, and water-related processes produced the encountered landforms and materials.
Nota Bene: A Bridge Too Far?
Authors: Smith Orla
Published: August 2004

His Brother's Keeper
A Story from the Edge of Medicine. by Jonathan Weiner. Ecco, New York, 2004. 368 pp. $26.95, C$36.95. ISBN 0-06-001007-X.

This account of one family's efforts to save a member from the fatal neurodegenerative disease amyotrophic lateral sclerosis also illuminates the promise and pitfalls of biomedical research.
Analysis of Clonal Composition of
Mycobacterium tuberculosis
Isolates in Primary Infections in Children
Authors: García de Viedma Darío , Marín Mercedes , Ruiz María Jesús , Bouza Emilio
Published: August 2004

The assumption that
Mycobacterium tuberculosis
infections should be considered clonally homogeneous has been weakened in the last few years. Recent studies have shown (i) the isolation of different
M. tuberculosis
strains from sequential episodes, (ii) mixed infections by two
M. tuberculosis
strains, and (iii) genetic variations in
M. tuberculosis
subpopulations due to microevolution events. Nevertheless, it is unknown whether clonal heterogeneity could be found in the initial steps of
M. tuberculosis
infection, i.e., the primary infection. In the present study we analyzed the clonal composition of the
M. tuberculosis
isolates causing primary infections in children. Cultures were clonally homogeneous in most cases (11 of 12). In 1 of the 12 cases (8.3%), clonal heterogeneity among the
M. tuberculosis
isolates was found by spoligotyping and IS
6110
-based restriction fragment length polymorphism analysis. This case occurred in a 2-year-old child in whom microevolution events were unlikely and who had no risk factors for overexposure to
M. tuberculosis
. Clonal heterogeneity should also be considered in primary
M. tuberculosis
infections, including circumstances in which it is usually unexpected.
Sex Pheromone Response, Clumping, and Slime Production in Enterococcal Strains Isolated from Occluded Biliary Stents
Authors: Donelli Gianfranco , Paoletti Claudia , Baldassarri Lucilla , Guaglianone Emilio , Di Rosa Roberta , Magi Gloria , Spinaci Cinzia , Facinelli Bruna
Published: August 2004

Bile-resistant bacteria, particularly gram-positive
Enterococcus faecalis
and
Enterococcus faecium
, play an important role in biliary stent occlusion, because their sessile mode of growth protects them against host defenses and antimicrobial agents. Twelve
E. faecalis
and seven
E. faecium
strains isolated from occluded biliary stents have been investigated for slime production, presence of aggregation substance genes, and ability to adhere to Caco-2 cells. Ten isolates were strong producers of slime, and seven isolates produced clumps when exposed to pheromones of
E. faecalis
JH2-2 and/or OG1RF. The small
E. faecium
clumps differed from the large clumps of
E. faecalis
and were similar to those of
E. faecium
LS10(pBRG1) carrying a pheromone response plasmid. After induction with pheromones, the adhesion to Caco-2 cells of clumping-positive strains was found to increase from two- to fourfold. Amplicons of the expected size were detected in three clumping-positive and three clumping-negative
E. faecalis
isolates by using primers (
agg
) internal to a highly conserved region of the
E. faecalis
pheromone response plasmids pAD1, pPD1, and pCF10 and primers internal to
prgB
of the
E. faecalis
plasmid pCF10. The
agg
/
prgB
-positive
E. faecalis
strains were also positive in Southern hybridization experiments with a
prgB
-specific probe. No PCR products were obtained with the same primers from four clumping-positive isolates (one
E. faecalis
and three
E. faecium
strains), which were also Southern hybridization negative. Our results demonstrate that slime production and pheromone response are both present in isolated enterococci, suggesting that clinical strains with these features might have a selective advantage in colonizing biliary stents.
Suicide PCR on Skin Biopsy Specimens for Diagnosis of Rickettsioses
Authors: Fournier Pierre-Edouard , Raoult Didier
Published: August 2004

As rickettsioses may be severe diseases andRickettsia prowazekiiis a potential agent of bioterrorism, highly efficient diagnostic techniques are required to detect rickettsiae in patients. We developed a nested PCR assay using single-use primers targeting single-use gene fragments present in the genomes of bothRickettsia conoriiandR. prowazekii. We used this “suicide” PCR with DNA from 103 skin biopsy specimens from patients who definitely had a rickettiosis, 109 skin biopsy specimens from patients who possibly had a rickettsiosis, and 50 skin biopsy specimens from patients with nonrickettsial diseases. The suicide PCR detected “R. conorii conorii” in 38 biopsy specimens,R. africaein 28 biopsy specimens,R. slovacain 12 biopsy specimens, “R. sibirica mongolotimonae” in 5 biopsy specimens,R. aeschlimanniiin 2 biopsy specimens, and “R. conorii caspia” and “R. sibirica sibirica” in 1 biopsy specimen each. The technique had a specificity of 100% and a sensitivity of 68%. It was 2.2 times more sensitive than culture (P< 10−2) and 1.5 times more sensitive than regular PCR (P< 10−2). The efficacy of the suicide PCR was reduced by antibiotic therapy prior to biopsy (P< 10−2) and was increased when it was performed with eschar biopsy specimens (P= 0.03). We propose the use of the suicide PCR as a sensitive, specific, and versatile technique for improving the diagnosis of rickettsioses, especially when it is used on eschar biopsy specimens taken prior to antibiotic therapy.
Characterization of a Trinucleotide Repeat Sequence (CGG)
5
and Potential Use in Restriction Fragment Length Polymorphism Typing of
Mycobacterium tuberculosis
Authors: Otsuka Yayoi , Parniewski Pawel , Zwolska Zofia , Kai Masatake , Fujino Tomoko , Kirikae Fumiko , Toyota Emiko , Kudo Koichiro , Kuratsuji Tadatoshi , Kirikae Teruo
Published: August 2004

The genomes of 28 bacterial strains, including mycobacterial species
Mycobacterium tuberculosis
and
Mycobacterium bovis
, were analyzed for the presence of a special class of microsatellite, that of trinucleotide repeat sequences (TRS). Results of a search of all 10 possible TRS motifs (i.e., CCT, CGG, CTG, GAA, GAT, GTA, GTC, GTG, GTT, and TAT) with five or more repeating units showed that (CGG)
5
was highly represented within the genomic DNA of
M. tuberculosis
and
M. bovis
. Most of the (CGG)
5
repeats in the genome were within the open reading frames of two large gene families encoding PE_PGRS and PPE proteins that have the motifs Pro-Glu (PE) and Pro-Pro-Glu (PPE). (CGG)
5
-probed Southern hybridization showed that some mycobacterial species, such as
Mycobacterium marinum
,
Mycobacterium kansasii
, and
Mycobacterium szulgai
, possess many copies of (CGG)
5
in their genomes. Analysis of clinical isolates obtained from Tokyo and Warsaw with both IS
6110
and (CGG)
5
probes showed that there is an association between the fingerprinting patterns and the geographic origin of the isolates and that (CGG)
5
fingerprinting patterns were relatively more stable than IS
6110
patterns. The (CGG)
5
repeat is a unique sequence for some mycobacterial species, and (CGG)
5
fingerprinting can be used as an epidemiologic method for these species as well as IS
6110
fingerprinting can. If these two fingerprinting methods are used together, the precise analysis of
M. tuberculosis
isolates will be accomplished. (CGG)
5
-based fingerprinting is particularly useful for
M. tuberculosis
isolates with few or no insertion elements and for the identification of other mycobacterial species when informative probes are lacking.
Characterization and Prevalence of MefA, MefE, and the Associated
msr
(D) Gene in
Streptococcus pneumoniae
Clinical Isolates
Authors: Daly Melissa M. , Doktor Stella , Flamm Robert , Shortridge Dee
Published: August 2004

Recent work has shown that the efflux genes in
Streptococcus pneumoniae
that are responsible for acquired macrolide resistance can be distinguished as either
mef
(E) or
mef
(A). The genetic elements on which
mef
(A) and
mef
(E) are found also carry an open reading frame (ORF) that is 56% homologous to
msr
(A) in
Staphylococcus
. The prevalence of
mef
(A/E) and of the
msr
-like ORF [
msr
(D)] was evaluated in 153

mef
+
S. pneumoniae

clinical isolates collected in North America, Europe, Africa, and Asia from 1997 to 2002. Clinical isolates were screened with PCR primers specific for either
mef
(A) or
mef
(E) and for
msr
(D).
mef
(A),
mef
(E), and
msr
(D) were cloned from

mef
+

strains and transformed into a susceptible, competent strain of
S. pneumoniae
. The transformants were tested for antimicrobial susceptibilities and efflux pump induction. The results of this work demonstrated that
mef
(A) is more often isolated in parts of Europe, with some incidence in Canada, and that the
msr
-like gene alone can confer the efflux phenotype.
Comparison of Results of Fluconazole Disk Diffusion Testing for
Candida
Species with Results from a Central Reference Laboratory in the ARTEMIS Global Antifungal Surveillance Program
Authors: Pfaller M. A. , Hazen K. C. , Messer S. A. , Boyken L. , Tendolkar S. , Hollis R. J. , Diekema D. J.
Published: August 2004

The accuracy of antifungal susceptibility tests is important for accurate resistance surveillance and for the clinical management of patients with serious infections. Our main objective was to compare the results of fluconazole disk diffusion testing of
Candida
spp. performed by ARTEMIS participating centers with disk diffusion and MIC results obtained by the central reference laboratory. A total of 2,949 isolates of
Candida
spp. were tested by NCCLS disk diffusion and reference broth microdilution methods in the central reference laboratory. These results were compared to the results of disk diffusion testing performed in the 54 participating centers. All tests were performed and interpreted following NCCLS recommendations. Overall categorical agreement between participant disk diffusion test results and reference laboratory MIC results was 87.4%, with 0.2% very major errors (VME) and 3.3% major errors (ME). The categorical agreement between the disk diffusion test results obtained in the reference laboratory with the MIC test results was similar: 92.8%. Likewise, good agreement was observed between participant disk diffusion test results and reference laboratory disk diffusion test results: 90.4%, 0.4% VME, and 3.4% ME. The disk diffusion test was especially reliable in detecting those isolates of
Candida
spp. that were characterized as resistant by reference MIC testing. External quality assurance data obtained by surveillance programs such as the ARTEMIS Global Antifungal Surveillance Program ensure the generation of useful surveillance data and result in the continued improvement of antifungal susceptibility testing practices.
Development of Two PCR-Based Techniques for Detecting Helical and Coccoid Forms of
Helicobacter pylori
Authors: Shahamat M. , Alavi M. , Watts J. E. M. , Gonzalez J. M. , Sowers K. R. , Maeder D. W. , Robb F. T.
Published: August 2004

The primary mode of transmission of
Helicobacter pylori
, a human pathogen carried by more than half the population worldwide, is still unresolved. Some epidemiological data suggest water as a possible transmission route.
H. pylori
in the environment transforms into a nonculturable, coccoid form, which frequently results in the failure to detect this bacterium in environmental samples by conventional culture techniques. To overcome limitations associated with culturing, molecular approaches based on DNA amplification by PCR have been developed and used for the detection of
H. pylori
in clinical and environmental samples. Our results showed the
glmM
gene as the most promising target for detection of
H. pylori
by PCR amplification. Under optimal amplification conditions,
glmM
-specific primers generated PCR-amplified products that were specific for
H. pylori
and some other
Helicobacter
species. Genome sequence analysis revealed the existence of a conserved region linked to a hypervariable region upstream of the 16S rRNA gene of
H. pylori
. Selective PCR primer sets targeting this sequence were evaluated for the specific detection of
H. pylori
. One primer set, Cluster2 and B1J99, were shown to be highly specific for
H. pylori
strains and did not produce any PCR products when other
Helicobacter
species and other bacterial species were analyzed. In tests with 32 strains of
H. pylori
, 6 strains of other
Helicobacter
species, 8 strains of
Campylobacter jejuni
, and 21 strains belonging to different genera, the primers for
glmM
were selective for the
Helicobacter
genus and the primers containing the region flanking the 16S rRNA gene were selective for
H. pylori
species only. The combination of two sensitive PCR-based methods, one targeting the
glmM
gene and the other targeting a hypervariable flanking region upstream of the 16S rRNA gene, are complementary to each other. Whereas the
glmM
-specific primers provide a rapid, sensitive presumptive assay for the presence of
H. pylori
and closely related
Helicobacter
spp., the primers for sequences flanking the 16S rRNA gene can confirm the presence of
H. pylori
and locate the potential source of this bacterium.
Rapid Identification of
Nocardia farcinica
Clinical Isolates by a PCR Assay Targeting a 314-Base-Pair Species-Specific DNA Fragment
Authors: Brown June M. , Pham Kim N. , McNeil Michael M. , Lasker Brent A.
Published: August 2004

Nocardia farcinica
is the most clinically significant species within the
Nocardia asteroides
complex. Differentiation of
N. farcinica
from other members of
N. asteroides
complex is important because this species characteristically demonstrates resistance to several extended-spectrum antimicrobial agents. Traditional phenotypic characterization of this species is time- and labor-intensive and often leads to misidentification in the clinical microbiology laboratory. We previously observed a 409-bp product for all strains of
N. farcinica
by using randomly amplified polymorphic DNA analysis with the primer DKU49. In this investigation, the 409-bp fragment was sequenced and then used to design a specific primer pair, Nf1 (16-mer) and Nf2 (16-mer), complementary to the 409-bp fragment. PCR amplification of genomic DNA from 28
N. farcinica
isolates with Nf1 and Nf2 generated a single intense 314-bp fragment. The specificity of the assay with these primers was verified, since there were no PCR amplification products observed from heterologous nocardial species (
n
= 59) or other related bacterial genera (
n
= 41). Restriction enzyme digestion using CfoI and direct sequencing of the 314-bp fragment further confirmed the specificity of the assay for
N. farcinica
. This highly sensitive and specific PCR assay provides a rapid (within 1 day of obtaining DNA) method for identification of this medically important emerging pathogen. Rapid diagnosis of
N. farcinica
infection may allow for earlier initiation of effective therapy, thus improving patient outcome.
Comparison of the Directigen Flu A+B Membrane Enzyme Immunoassay with Viral Culture for Rapid Detection of Influenza A and B Viruses in Respiratory Specimens
Authors: Cazacu Andreea C. , Chung Sooyoung E. , Greer Jewel , Demmler Gail J.
Published: August 2004

The performance of a commercially available, rapid membrane enzyme immunoassay for influenza A and B virus detection was compared to that of viral culture in 4,092 respiratory specimens collected from patients presenting with respiratory symptoms during the 2002-2003 influenza season. The test's overall sensitivity was 43.83%, lower than previously reported but similar for detection of both influenza A and B viruses (42.98 versus 44.76%). However, specificity, 99.74%, was excellent for both influenza A and B viruses (99.82 versus 99.92%). These values make this test a very good confirmatory test when clinical suspicion is high, but a less accurate screening test for large populations.
Concurrent Infections with Vector-Borne Pathogens Associated with Fatal Hemolytic Anemia in a Cattle Herd in Switzerland
Authors: Hofmann-Lehmann Regina , Meli Marina L. , Dreher Ute M. , Gönczi Enikö , Deplazes Peter , Braun Ueli , Engels Monika , Schüpbach Jörg , Jörger Kaspar , Thoma Rudolf , Griot Christian , Stärk Katharina D. C. , Willi Barbara , Schmidt Joseph , Kocan Katherine M. , Lutz Hans
Published: August 2004

Bovine anaplasmosis is a vector-borne disease that results in substantial economic losses in other parts of the world but so far not in northern Europe. In August 2002, a fatal disease outbreak was reported in a large dairy herd in the Swiss canton of Grisons. Diseased animals experienced fever, anorexia, agalactia, and depression. Anemia, ectoparasite infestation, and, occasionally, hemoglobinuria were observed. To determine the roles of vector-borne pathogens and to characterize the disease, blood samples were collected from all 286 animals: 50% of the cows were anemic. Upon microscopic examination of red blood cells,Anaplasma marginaleinclusion bodies were found in 47% of the cows. The infection was confirmed serologically and by molecular methods. Interestingly, we also found evidence of infections withAnaplasma phagocytophilum, largeBabesiaandTheileriaspp., andMycoplasma wenyonii. The last two species had not previously been described in Switzerland. Anemia was significantly associated with the presence of the infectious agents detected, with the exception ofA. phagocytophilum. Remarkably, concurrent infections with up to five infectious vector-borne agents were detected in 90% of the ill animals tested by PCR. We concluded thatA. marginalewas the major cause of the hemolytic anemia, while coinfections with other agents exacerbated the disease. This was the first severe disease outbreak associated with concurrent infections with vector-borne pathogens in alpine Switzerland; it was presumably curtailed by culling of the entire herd. It remains to be seen whether similar disease outbreaks will have to be anticipated in northern Europe in the future.